Macrocyclic NS3-serine protease inhibitors of hepatitis C virus comprising N-cyclic P2 moieties

FIELD OF INVENTION

BACKGROUND OF THE INVENTION

[00009] HCV has been implicated in cirrhosis of the liver and in induction of hepatocellular carcinoma. The prognosis for patients suffering from HCV infection is currently poor. HCV infection is more difficult to treat than other forms of hepatitis due to the lack of immunity or remission associated with HCV infection. Current data indicates a less than 50% survival rate at four years post cirrhosis diagnosis. Patients diagnosed with localized resectable hepatocellular carcinoma have a five-year survival rate of 10-30%, whereas those with localized unresectable hepatocellular carcinoma have a five-year survival rate of less than Reference is also made to WO 00/09558 (Assignee: Boehringer Ingelheim Limited; Published Feb. 24, 2000) which discloses peptide derivatives of the formula:
where the various elements are defined therein. An illustrative compound of that series is:
Reference is also made to WO 00/09543 (Assignee: Boehringer Ingelheim Limited; Published Feb. 24, 2000) which discloses peptide derivatives of the formula:
where the various elements are defined therein. An illustrative compound of that series is:

SUMMARY OF THE INVENTION

[00019] In its principal embodiment, the present invention provides a macrocyclic compound of Formula I:
wherein:

• • [00021] X and Y are independently selected from the moieties: alkyl, alkyl-aryl, heteroalkyl, heteroaryl, aryl-heteroaryl, alkyl-heteroaryl, cycloalkyl, alkyl ether, alkyl-aryl ether, aryl ether, alkyl amino, aryl amino, alkyl-aryl amino, alkyl sulfide, alkyl-aryl sulfide, aryl sulfide, alkyl sulfone, alkyl-aryl sulfone, aryl sulfone, alkyl-alkyl sulfoxide, alkyl-aryl sulfoxide, alkyl amide, alkyl-aryl amide, aryl amide, alkyl sulfonamide, alkyl-aryl sulfonamide, aryl sulfonamide, alkyl urea, alkyl-aryl urea, aryl urea, alkyl carbamate, alkyl-aryl carbamate, aryl carbamate, alkyl-hydrazide, alkyl-aryl hydrazide, alkyl hydroxamide, alkyl-aryl hydroxamide, alkyl sulfonyl, aryl sulfonyl, heteroalkyl sulfonyl, heteroaryl sulfonyl, alkyl carbonyl, aryl carbonyl, heteroalkyl carbonyl, heteroaryl carbonyl, alkoxycarbonyl, aryloxycarbonyl, heteroaryloxycarbonyl, alkylaminocarbonyl, arylaminocarbonyl, heteroarylaminocarbonyl or a combination thereof, with the proviso that X and Y may optionally be additionally substituted with moieties selected from the group consisting of aromatic, alkyl, alkyl-aryl, heteroalkyl, aryl-heteroaryl, alkyl-heteroaryl, cycloalkyl, alkyl ether, alkyl-aryl ether, alkyl sulfide, alkyl-aryl sulfide, alkyl sulfone, alkyl-aryl sulfone, alkyl amide, alkyl-aryl amide, alkyl sulfonamide, alkyl amines, alkyl-aryl amines, alkyl-aryl sulfonamide, alkyl urea, alkyl-aryl urea, alkyl carbamate and alkyl-aryl carbamate;
  • [00022] R¹═COR⁵ or B(OR)₂, wherein R⁵═H, OH, OR⁸, NR⁹R¹⁰, CF₃, C₂F₅, C₃F₇, CF₂R⁶, R⁶, COR⁷ wherein R⁷═H, OH, OR⁸, CHR⁹R¹⁰, or NR⁹R¹⁰, wherein R⁶, R⁸, R⁹ and R¹⁰ are independently selected from the group consisting of H, alkyl, aryl, heteroalkyl, heteroaryl, cycloalkyl, cycloalkyl, arylalkyl, heteroarylalkyl, CH(R¹′)COOR¹¹, CH(R¹′)CONR¹²R¹³, CH(R¹′)CONHCH(R²′)COO R¹¹, CH(R¹′)CONHCH(R²′)CONR¹²R¹³, CH(R¹′)CONHCH(R²)R′, CH(R¹′)CONHCH(R²′)CONHCH(R³′)COO R¹¹, CH(R¹′)CONHCH(R²′)CONHCH(R³′)CONR¹²R¹³, CH(R¹′)CONHCH(R²′)CONHCH(R³′)CONHCH(R⁴′)COO R¹¹, CH(R¹′)CONHCH(R²′)CONHCH(R³′)CONHCH(R⁴′)CONR¹²R¹³, CH(R¹′)CONHCH(R²′)CONHCH(R³′)CONHCH(R⁴′)CONHCH(R⁵′)COO R¹¹, CH(R¹′)CONHCH(R²′)CONHCH(R³′)CONHCH(R⁴′)CONHCH(R⁵′) CONR¹²R¹³, wherein R¹′, R²′, R³′, R⁴′, R⁵′, R¹¹, R¹², R¹³, and R′ are independently selected from a group consisting of H, alkyl, aryl, heteroalkyl, heteroaryl, cycloalkyl, alkyl-aryl, alkyl-heteroaryl, aryl-alkyl and heteroaralkyl;
  • [00023] Z is selected from O, N, or CH;
  • [00024] W may be present or absent, and if W is present, W is selected from C═O, C═S, or SO₂;
  • [00025] Q maybe present or absent, and when Q is present, Q is CH, N, P, (CH₂)_p, (CHR)_p, (CRR′)_p, O, NR, S, or SO₂; and when Q is absent, M is also absent, and A is directly linked to X;
  • [00026] A is O, CH₂, (CHR)_p, (CHR-CHR′)_p, (CRR′)_p, NR, S, SO₂ or a bond;
  • [00027] E is CH, N or CR, or a double bond towards A, L or G;
  • [00028] G may be present or absent, and when G is present, G is (CH₂)_p, (CHR)_p, or (CRR′)_p′; and when G is absent, J is present and E is directly connected to the carbon atom where G was connected to;
  • [00029] J maybe absent or present, and when J is present, J is (CH₂)_p, (CHR)_p, or (CRR′)_p, SO₂, NH, NR or O; and when J is absent, G is present and E is directly linked to N;
  • [00030] L may be present or absent, and when L is present, L is CH, CR, O, S or NR; and when L is absent, then M may be absent or present, and if M is present with L being absent, then M is directly and independently linked to E, and J is directly and independently linked to E;
  • [00031] M may be present or absent, and when M is present, M is O, NR, S, SO₂, (CH₂)_p, (CHR)_p (CHR-CHR′)_p, or (CRR′)_p;
  • [00032] p is a number from 0 to 6; and
  • [00033] R, R′, R², R³ and R⁴ are independently selected from the group consisting of H; C1-C10 alkyl; C2-C10 alkenyl; C3-C8 cycloalkyl; C3-C8 heterocycloalkyl, alkoxy, aryloxy, alkylthio, arylthio, amino, amido, ester, carboxylic acid, carbamate, urea, ketone, aldehyde, cyano, nitro; oxygen, nitrogen, sulfur, or phosphorus atoms with said oxygen, nitrogen, sulfur, or phosphorus atoms numbering zero to six;
    (cycloalkyl)alkyl and (heterocycloalkyl)alkyl, wherein said cycloalkyl is made of three to eight carbon atoms, and zero to six oxygen, nitrogen, sulfur, or phosphorus atoms, and said alkyl is of one to six carbon atoms; aryl; heteroaryl; alkyl-aryl; and alkyl-heteroaryl;
    with said alkyl, heteroalkyl, alkenyl, heteroalkenyl, aryl, heteroaryl, cycloalkyl and heterocycloalkyl moieties may be optionally substituted, with said term “substituted” referring to optional and suitable substitution with one or more moieties selected from the group consisting of alkyl, alkenyl, alkynyl, aryl, aralkyl, cycloalkyl, heterocyclic, halogen, hydroxy, thio, alkoxy, aryloxy, alkylthio, arylthio, amino, amido, ester, carboxylic acid, carbamate, urea, ketone, aldehyde, cyano, nitro, sulfonamide, sulfoxide, sulfone, sulfonyl urea, hydrazide, and hydroxamate.

DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS

[00048] In one embodiment, the present invention discloses compounds of Formula I as inhibitors of HCV protease, especially the HCV NS3/NS4a serine protease. Among the compounds encompassed by Formula I, preferred compounds are those which have the Formula II:
or a pharmaceutically acceptable derivative thereof, where the various definitions are given above. Some of the preferred embodiments include, but are not limited to, the following definitions of the various functionalities in the above-noted general formulas I and II. Thus, for example, R² in formula I may be selected from the following moieties:
Some preferred representations for the moiety
are, for example, the following structures a, b, or c:
The structure a may be selected from the following non-limiting types of structures:
Other additional preferred embodiments are, for example, where the moiety:
is sometimes the following structures, where the definitions for the various positions are exemplified in the structures of compounds shown later in this section:
In some more preferred compounds, the moieties G and J are independently selected from (CH₂)_p, (CHR)_p, (CHR-CHR′)_p, and (CRR′)_p, and the moiety A-E-L-M-Q is an aromatic ring consisting of two to eight carbon atoms, zero to six hetero atoms with X and J being ortho, para or meta with respect to each other.
In other preferred embodiments, R³ in formula I is selected from the following structures:
wherein

• • [00058] R³⁰═H, CH₃ or other alkyl groups;
  • [00059] R³¹═OH, O-alkyl, NH₂ or N-alkyl;
  • [00060] R³² and R³³ may be the same or different and are independently selected from H,
  • [00061] F, Cl, Br and CH₃;
  • [00062] and the moiety X-Y is selected from the following structures:

[00065] Some of the inventive compounds and the methods of synthesizing the various types of the inventive compounds are listed below, then schematically described, followed by the illustrative Examples.

[00133] The preparation of the compound of formula 1h, wherein R¹, R², R³ are defined above, R′ is alkyl, heteroalkyl (OR″, SR′″, NR″R′″ wherein R″ and R′″ are alkyl groups), halo substituent at ortho, meta, or para -position to oxygen atom; R is alkyl, aryl, or alkylaryl groups; n is from zero to five; X is (CH₂)_m where m is one to five, oxygen atom, NY where Y is hydrogen atom, alkyl, aryl group; and PG¹ and PG² are appropriate protecting groups (PG¹=t-boc, cbz and PG²═H, Bn etc) is outlined in Scheme 1. The protected 4-hydroxyproline acid (1a) is alkylated at the 4-position by an alkyl bromide in the presence of sodium hydride. The product 1b is then converted to an ester either with an alcohol under acidic conditions, or with trimethylsilyldiazamethane. After deprotection, the resulting amine is coupled to a Boc-protected amino acid in the presence of HOOBt, EDCl.HCl and NMM. After removal of the Boc group from product 1d, the dipeptide is reacted with a substituted hydroxyphenyl acetic acid using the same coupling conditions. Catalytic hydrogenation of the benzyl ether gives the precursor for the macrocyclization. The macrocyclization is achieved under Mitsunobu conditions by using triphenylphosphine and ADDP. (The Mitsunobu reaction is reviewed by D. L. Hughes, Org. Reactions, 42 (1992) 335, John Wiley & Sons, New York, L. Paquette, ed.) After the ester is hydrolyzed to an acid with lithium hydroxide, it is coupled to an amine intermediate to afford 1h.

[00134] The preparation of the compound of Formula 2e, wherein R¹, R², R³, R and R′ are defined above, is outlined in Scheme 2. The protected 3,4-dehydroproline 2a is diastereoselectively dihydroxylated to afford cis-diol 2b. The acetal formation between 2b and an aldehyde can be accomplished in the presence of catalytic amount of p-toluenesulfonic acid. The bicyclic proline derivative 2c is converted to the macrocyclic ester 2d and subsequently to HCV inhibitor 2e according to the sequence outlined in Scheme 1.

[00135] The preparation of the compound of Formula 3f, wherein R¹, R², R³, R, R′ and n are defined above, is outlined in Scheme 3. When treated with trifluoroboron diethyletherate, the protected 4-hydroxyproline 3a and alkene 3b are converted to proline ether 3c, which undergoes the same sequence of transformations outlined in Scheme 1 to give the macrocyclic ester 3e and subsequently, to the desired final product 3f.

[00136] The preparation of the compound of Formula 4f, wherein R¹, R², R³, R, and n are defined above, is outlined in Scheme 4. The coupling between amino ester 4a and a Boc-protected amino acid was realized in the presence of NMM, HOOBt, and EDCl.HCl. After removal of the Boc group from 4b, the dipeptide is coupled to a terminal alkene carboxylic acid to give 4c. The double bond was then converted to an alcohol through hydroboration. The macrocyclization of the phenol alcohol is accomplished under Mitsunobu conditions by using triphenylphosphine and ADDP. After the resulting ester 4e is hydrolyzed to an acid with lithium hydroxide, the acid is coupled to an amine intermediate to afford 4f.

[00137] The preparation of the compound of Formula 5h in Scheme 5 where in R, R², R³, R′ are defined in Scheme 1, R⁴ being alkyl, cycloalkyl, aryl, heteroaryl, and heteroalkyl and R⁵ is OR, NR₂, or OH. The compound 5b was obtained by a Wittig reaction of 5a with tert-butyl phosphonoacetate and NaH. The compound 5b was converted to 5c by the treatment of MCPBA. The epoxide 5c was further opened with NaN₃ to give compound 5d which was reduced with Pd/C/H₂ to the amine and Cbz protected using Cbz-Cl, Et₃N to obtain compound of the formula 5e. The compound 5e was deprotected with TFA and further elaborated to 5f. The Cbz group of 5f was hydrogenolyzed and then coupled with compound of formula 1g using EDCl, HOOBt, NMM to obtain 5g. The compound of the type 5g is oxidized with Dess-Martin reagent to generate compounds of formula 5h.

[00138] The compound of formula 6m is synthesized as outlined in Scheme 6 wherein R¹, R², R′ and n are defined in Scheme 1 and R⁶ being alkyl, aryl, ester, carboxylic acid and carboxylamides. The compound of type 6b is synthesized from 6a by a Wittig olefination using Ph₃PCH₃l and BuLi. The compound 6b is further aminohydroxylated to synthesize compound of the type 6c, which was reduced using Rh/C, and H₂ to afford compound of type 6d. The compound 6d was oxidized to compound of type 6e using RuCl₃ and H₅lO₆. The compound 6e was elaborated to compound 6i by coupling it with deprotected 6h using NMM, EDCl, and HOOBt. Extension of compound 6i to 6j was again achieved by coupling deprotected 6i and appropriately substituted phenyl acetic acid using EDCl, HOOBt and NMM. The compound 6k obtained after hydrogenolysis of benzyl group in 6j was cyclized to 6l using the Mitsunobu conditions. 6l was further elaborated to compounds of type 6m as outlined in Scheme 1.

[00139] The compounds of type 7d are synthesized using the arene ruthenium chemistry wherein the substituents R¹, R2, R³, R′ are defined in Scheme 1, and n=0 to 3. The synthesis of the compound of formula 7b was obtained from compound of type 7a by an EDCl, HOBt, Hünigs base coupling. The treatment of 7b with Cs₂CO₃ and photolytic removal of ruthenium converted the compound of type 7b to type 7c, which was further elaborated to compounds 7d as outlined in Scheme 1.

[00140] The preparation of the compound of formula 8e, wherein R¹, R², R³ are defined above, R′ is alkyl, heteroalkyl (OR″, SR′″, NR″R′″ wherein R″ and R′″ are alkyl groups), halo substituent at ortho, meta, or para-position to oxygen atom; R is alkyl, aryl, or alkylaryl groups; n is from zero to five; and X is (CH₂)_m where m is one to five, oxygen atom, NY where Y is hydrogen atom, alkyl, aryl group is outlined in Scheme 8. The protected cis-4-hydroxyproline derivative 8a is converted to the brosylate (Bs=4-bromobenzenesulfonyl) derivative 8b which was displaced with the appropriate mercaptoalcohol under sodium hydride conditions to give 8c. Conversion to the macrocyclic ester 8d and subsequently to the desired target 8e was accomplished as outlined in Scheme 1.

[00141] The preparation of the compound of formula 9d, wherein R¹, R², R³ are defined above, R′ is alkyl, heteroalkyl (OR″, SR′″, NR″R′″ wherein R″ and R′″ are alkyl groups), halo substituent at ortho, meta, or para-position to oxygen atom; R is alkyl, aryl, or alkylaryl groups; n is from zero to five; X is (CH₂)_m where m is one to five, oxygen atom, NY where Y is hydrogen atom, alkyl, aryl group; and LG is leaving group (e.g., OTs, Br) is outlined in Scheme 9. The protected 4-hydroxyproline derivative 1a is converted to 9a via formation of 11d (which is described in Scheme 1). Removal of the benzyl ether followed by conversion to an appropriate leaving group and unravelling the N protecting group provided 9b. Conversion to the macrocyclic ester 9c was carried out by treatment with sodium carbonate/sodium iodide in refluxing acetone. Subsequent processing to the desired target 9d was accomplished as outlined in Scheme 1.

[00142] The preparation of the compound of formula 10e, wherein R¹, R², R³ are defined above, R′ is alkyl, heteroalkyl (OR″, SR′″, NR″R′″ wherein R″ and R′″ are alkyl groups), halo substituent at ortho, meta, or para -position to oxygen atom; R is alkyl, aryl, or alkylaryl groups; n and q is any combination of zero to five; V is oxygen atom, NY where Y is hydrogen atom, alkyl, aryl group; W is alkyl, aryl, alkylaryl, heteroaryl; PG′ and PG² are appropriate protecting groups (PG¹=t-boc, cbz and PG²═H, Bn, etc) is outlined in Scheme 10. The protected 4-hydroxyproline derivative 1a is converted to 10b as described in Scheme 1. Removal of the protecting group (10c) followed by treatment with phosgene equivalent results in the macrocyclic ester 10d. Conversion to the desired target 10e was accomplished as outlined in Scheme 1.

[00143] The preparation of the compound of formula 11g, wherein R¹, R², R³ are defined above, R is alkyl, aryl, or alkylaryl groups; X is (CH₂)_m, (CH₂)_mO, (CH₂)_mNY where m is one to five, and Y is hydrogen atom, alkyl, aryl group; A is hydrogen atom or appropriately positioned halogen atom; PG¹ is appropriate protecting groups (PG¹=t-boc, cbz, etc) is outlined in Scheme 11. The protected 4-aminoproline derivative 11a is converted to 11b by treatment with suitable benzenesulfonyl chloride and a base. Removal of protecting group and coupling with a protected aminoacid derivative gave 11c. This was converted to 11d using similar deprotection, coupling strategy. Palladium(0) catalyzed cyclization afforded 11e as a mixture of isomers which was hydrogenated to provide macrocyclic ester 11f. Subsequent conversion to the desired target 11g was accomplished as outlined in Scheme 1.

[00144] The preparation of the compound of formula 12d, wherein R¹, R², R³ are defined above, R is alkyl, aryl, or alkylaryl groups; n is from zero to five; X is (CH₂)_m where m is one to five, oxygen atom, NY where Y is hydrogen atom, alkyl, aryl group; A is hydrogen atom or appropriately positioned halogen atom; and PG¹ and PG² are appropriate protecting groups (PG¹=t-boc, cbz and PG²═H, Bn etc) is outlined in Scheme 12. The protected 4-aminoproline derivative 11a is converted to 11c as described in Scheme 11. Conversion of 11c to the macrocyclic ester 12c and subsequent conversion to the desired target 12d was accomplished as outlined in Scheme 1.
Preparation of Intermediates
Intermediate A

[00147] To a stirred solution of 1-nitrobutane (16.5 g, 0.16 mol) and glyoxylic acid in H₂O (28.1 g, 0.305 mol) and MeOH (122 mL) at 0° C.-5° C., was added dropwise triethyl amine (93 mL, 0.667 mol) over 2 hrs. The solution was warmed to room temperature, stirred overnight and concentrated to dryness to give an oil. The oil was then dissolved in H₂O and acidified to pH=1 with 10% HCl, followed by extraction with EtOAc. The combined organic solution was washed with brine, dried over Na₂SO₄, filtered and concentrated to dryness to give the product ii (28.1 g, 99% yield)

[00148] To a stirred solution of starting material ii (240 g, 1.35 mol) in acetic acid (1.25 L) was added 10% Pd/C (37 g). The resulting solution was hydrogenated at 59 psi for 3 hrs and then at 60 psi overnight. The acetic acid was then evaporated and azeotroped 3 times with toluene, then triturated with MeOH and ether. The solution was then filtered and azeotroped twice with toluene to give an off white solid (131 g, 0.891 mol, 66%).

[00149] To a stirred solution of the amino acid iii (2.0 g, 13.6 mmol) in dioxane (10 mL) and H₂O (5 mL) at 0° C., was added 1N NaOH solution (4.3 mL, 14.0 mmol). The resulting solution was stirred for 10 minutes, followed by addition of di-t-butyldicarbonate (0.110 g, 14.0 mmol) and stirred at 0° C. for 15 minutes. The solution was then warmed to room temperature, stirred for 45 minutes and kept at refrigerator overnight and concentrated to dryness to give a crude material. To the solution of this crude material in EtOAc (100 mL) and ice, was added KHSO₄ (3.36 g) and H₂O (32 mL) and stirred for 4-6 minutes. The organic layer was then separated and the aqueous layer was extracted twice with EtOAc and the combined organic layer was washed with water, brine, dried over Na₂SO₄, filtered and concentrated to dryness to give the product as a clear gum (3.0 g, 89% yield).

[00150] To a stirred solution of starting material (3.00 g, 12.0 mmol) in DMF (15 mL) and CH₂Cl₂ (15 mL) at −20° C. was added HOOBt (1.97 g, 12.0 mmol), N-methyl morpholine (4.0 mL, 36.0 mmol) and EDCl (2.79 g, 14.5 mmol) and stirred for 10 minutes, followed by addition of HCl.H₂N-Gly-OBn (2.56 g, 13.0 mmol). The resulting solution was stirred at −20° C. for 2 hrs, then kept at refrigerator overnight and concentrated to dryness, followed by dilution with EtOAc (150 mL). The EtOAc solution was then washed twice with saturated NaHCO₃, H₂O, 5% H₃PO₄, brine, dried over Na₂SO₄, filtered and concentrated to dryness to give the product (4.5 g, 94%). LRMS m/z MH⁺=395.1.

[00151] The solution of starting material v (7.00 g, 17.8 mmol) in absolute ethanol (300 mL) was stirred at room temperature under a hydrogen atmosphere in the presence of Pd-C (300 mg, 10%). The reaction progress was monitored by TLC. After 2h, the mixture was filtered through a celite pad and the resulting solution was concentrated in vacuo to give the product vi (5.40 g, quantitative). LRMS m/z MH⁺=305.1.

[00152] To a solution of dimethylamine hydrochloride (1.61 g, 19.7 mmol), N-Boc-phenylglycine (4.50 g, 17.9 mmol), HOOBt (3.07 g, 18.8 mmol) and EDCl (4.12 g, 21.5 mmol) in anhydrous DMF (200 mL) and CH₂Cl₂ (150 mL) at −20° C. was added NMM (5.90 mL, 53.7 mmol). After being stirred at this temperature for 30 min, the reaction mixture was kept in a freezer overnight (18 h). It was then allowed to warm to rt., and EtOAc (450 mL), brine (100 mL) and 5% H₃PO₄ (100 mL) were added. After layers were separated, the organic solution was washed with 5% H₃PO₄ (100 mL), saturated aqueous sodium bicarbonate solution (2×150 mL), water (150 mL), and brine (150 mL), dried (MgSO₄), filtered and concentrated in vacuo to afford crude product viii (4.86 g) as a white solid, which was used without further purification.

[00153] The N-Boc-phenylglycine dimethylamide viii (4.70 g, crude) was dissolved in 4 N HCl (60 mL, 240 mmol) and the resulting solution was stirred at room temperature. The progress of the reaction was monitored by TLC. After 4 h, the solution was concentrated in vacuo to yield a white solid which was used in the next reaction without further purification. LRMS m/z MH⁺=179.0.

[00154] The desired compound x was prepared according to the coupling procedures described in Step 4. LRMS m/z MH⁺=465.1.

[00155] The desired intermediate A was prepared from tripeptide x according to the procedures described in Step 7. LRMS m/z MH⁺=365.1.
Intermediate B

[00157] The desired product xii was obtained by the procedure described for Intermediate A, Step 8 using commercially available xi as the coupling partner. The crude material was sufficiently pure for further studies. A portion of the product was purified by flash chromatography using 97/3 dichloromethane/MeOH. HRMS (FAB) Calcd for C₂₅H₄₀N₃O₇: 494.2866 (M+H)⁺. Found: 494.2863.

[00158] The desired product B was obtained by the procedure described for Intermediate A, Step 7. The crude material was used without further purification.
Intermediate C

[00160] The desired compound xiii was prepared similar to the coupling procedures described in Step 6 for intermediate A except for substituting methylamine for dimethylamine.

[00161] The desired compound xiv was prepared from xiii according to the procedures described in Step 7 for intermediate A.

[00162] The desired compound xv was prepared according to the coupling procedures described in Step 6 for intermediate A, except for substituting amine xiv for amine ix. LRMS m/z MH⁺=451.1.

[00163] The desired intermediate C was prepared according to the procedures described in Step 7 for intermediate A. LRMS m/z MH⁺=351.1. It was used without further purification.
Intermediate D

[00165] The desired intermediate D was prepared from compound v according to the procedures described in Step 7 for intermediate A. It was used without further purification.
Intermediate E

[00167] To a solution of xvi (5g) in dichloromethane (20 mL) was added TFA (20 ML) and stirred at ambient temperature for 4 hrs. Another portion of TFA (10 mL) was added and left standing for additional 3 hrs. All the volatiles were evaporated and it was dried in vacuo to provide quantitative yield of xvii. This material was carried further. (Note: The starting material xvi was obtained by a similar protocol described for B, using nitropentane as precursor).

[00168] The desired compound xviii was obtained by the method described for Intermediate A, Step 3. Sodium carbonate was used as the base instead of NaOH. The crude material xviii was carried further without purification.

[00169] To a cold (−20° C.) solution of xviii (4.8 g, 10.7 mmol) in dichloromethane/DMF (4/1, 25 mL) was added dimethylamine hydrochloride (959 mg, 11.77 mmol), diisopropylethylamine (4.2 mL, 24.1 mmol) and BOP (6.14 g, 13.89 mmol). It was left standing at −8° C. overnight. The reaction mixture was diluted with dichloromethane and washed with 10% citric acid solution, saturated NaHCO₃ solution and brine. The organic layer was dried (Na₂SO₄) and concentrated. The residue was purified by flash column chromatography using 97.5/2.5 dichloromethane/MeOH to afford 2.4 g of xix (47% yield). HRMS (FAB) Calcd for C₂₄H₃₉N₄O₆: 479.2870 (M+H)⁺. Found: 479.2862.

[00170] The desired product E was obtained by the procedure described for Intermediate A, Step 7. The crude material was used without further purification.
Intermediate F

[00172] To a cold (0° C.) solution of xx (20.0 g, 86.5 mmol) in benzene (300 mL) and MeOH (50 mL) was added trimethylsilyl diazomethane (2M in hexanes, 56 mL, 112 mmol) dropwise till the solution remained yellow. The reaction mixture was concentrated to provide 21 g of xxi (99% yield) which was pure enough to be carried further. HRMS (FAB) Calcd for C₁₁H₂₀NO₅: 246.1341 (M+H)⁺. Found: 246.1347.

[00173] To a cold (˜5° C.) mechanically stirred mixture of triphenylphosphine (31.97 g, 121.9 mmol) and methanesulfonic acid (7.66 mL, 118.1 mmol) in toluene was added DEAD (26.47 g, 152 mmol) slowly while maintaining the reaction temperature below 35° C. After the addition was complete, the reaction mixture was cooled to 20° C. and a solution of xxi (23.71 g, 96.8 mmol) in toluene was added followed by triethylamine (5.39 mL, 38.7 mmol). The mixture was heated to 70-75° C. for 6 hrs and cooled to 5-10° C. for 1 hr. All the solid material were filtered off and the filtrate was washed with 5% KH₂PO₄ solution and brine. The organic layer was dried (Na₂SO₄), and concentrated. Flash column chromatography of the residue using 95/5 dichloromethane/EtOAc provided 26 g of xxii (83% yield). HRMS (FAB) Calcd for C₁₂H₂₂NO₇S: 324.1117 (M+H)⁺. Found: 324.1115.

[00174] The mesylate xxii (26 g, 80.4 mmol) was dissolved in DMF and all the volatiles were evaporated in vacuo. (CAUTION: Trace amounts of dichloromethane must be removed by this process). To the remaining solution was added sodium azide (5.75 g, 88.4 mmol) and warmed to 70° C. over 5 hrs. The reaction mixture was cooled, diluted with EtOAc and washed with saturated NaHCO₃. The organic layer was dried (Na₂SO₄) and concentrated to afford 18 g (83% yield) of xxiii which was sufficiently pure for further studies.

EXAMPLES

Example 1

Preparation of Compounds of Formulas 1A and 1B

[00176]

[00177] To a solution of Boc-Hyp-OH (7.0 g, 30.3 mmol) and benzyl 3-bromopropyl ether (7.8 g, 34.0 mmol) in anhydrous DMF (400 mL) at room temperature was added sodium hydride (3.5 g, 60% dispersion in mineral oil, 87.5 mmol) and sodium iodide (0.5 g, 3.33 mmols) with stirring. The resulting suspension was vigorously stirred at room temperature overnight (18 h). The reaction was quenched carefully with a slow addition of water (50 mL) and acidified with 6 N HCl solution (20 mL). After addition of ethyl acetate (800 mL), brine (150 mL) and more water (150 mL), the formed two layers were separated and the organic layer was washed with 5% H₃PO₄ (3×200 mL). It was then dried with MgSO₄, filtered and concentrated in vacuo to afford I b as an oil which was used in Step B without further purification.

[00178] The acid 1b from Step A was dissolved in benzene (25 mL) and methanol (28 mL). To this solution at room temperature was added a solution of trimethylsilyl diazomethane (27 mL, 2.0 M in cyclohexane) with caution. After being stirred at room temperature for 1 h, it was concentrated in vacuo to yield the methyl ester. Flash chromatography (8 to 20% EtOAc-CH₂Cl₂) afforded 1c (5.15 g; 13.1 mmol, 43%, 2 steps) as an oil.

[00179] The Boc-amino methyl ester 1c (5.83 g, 14.8 mmol) was dissolved in 4 N HCl in dioxane (80 mL, 320 mmol) and the resulting solution was stirred at room temperature. The progress of the reaction was monitored by TLC. After 5 h, the solution was concentrated in vacuo and the residue was kept under vacuum overnight to yield a white solid which was used in the next coupling reaction without further purification.

[00180] To a solution of the amine ester 1d (from Step 1B), N-Boc-cyclohexylglycine (4.10 g, 14.9 mmol), HOOBt (2.60 g, 15.9 mmol) and EDCl (3.41 g, 17.8 mmol) in anhydrous DMF (150 mL) and CH₂Cl₂ at −20° C., was added NMM (6.50 mL, 59.1 mmol). After being stirred at this temperature for 30 min, the reaction mixture was kept in a freezer overnight (18 h). It was then stirred in air and allowed to warm to room temperature in 1 h. EtOAc (450 mL), brine (100 mL) and 5% H₃PO₄ (100 mL) were added. The separated organic solution was washed with 5% H₃PO₄ (100 mL), saturated aqueous sodium bicarbonate solution (2×150 mL), water (150 mL), and brine (150 mL), dried with magnesium sulfate, filtered and concentrated in vacuo. Flash chromatography (10 to 20% EtOAc-CH₂Cl₂) afforded 1e (6.60 g, 84% 2 steps) as a white solid. ¹H NMR (400 MHz, d₆-DMSO) δ 7.36-7.25 (m, 5H), 6.87 (d, J=8.9 Hz, 1H), 4.46-4.40 (m, 2H), 4.25 (t, J=8.3 Hz, 1H), 4.11 (s, 1H), 4.05-4.04 (m, 1H), 4.03-3.94 (m, 1H), 3.60 (s, 3H), 3.50-3.41 (m, 4H), 2.25-2.20 (m, 1H), 1.95-1.88 (m, 1H), 1.77-1.55 (m, 9H), 1.35 (s, 9H), 1.19-0.90 (m, 4H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.0, 170.7, 155.6, 138.8, 128.2, 127.4, 127.3, 78.0, 77.1, 71.9, 66.6, 65.1, 57.4, 56.3, 51.8, 34.5, 29.6, 28.6, 28.2, 25.9, 25.6, 25.5.

[00181] The Boc-amino methyl ester 1e (6.53 g, 12.3 mmol) was dissolved in 4 N HCl (60 mL, 240 mmol) and the resulting solution was stirred at room temperature. The progress of the reaction was monitored by TLC. After 4 h, the solution was concentrated in vacuo and the residue was kept under vacuum overnight to give a white solid which was used in the next coupling reaction without further purification. ¹H NMR (400 MHz, d₆-DMSO) δ 7.36-7.27 (m, 5H), 4.43 (s, 2H), 4.35-4.31 (m, 1H), 3.88 (d, J =11.7 Hz, 1H), 3.62 (s, 3H), 3.62-3.57 (m, 2H), 3.53-3.41 (m, 3H), 2.32-2.27 (m, 1H), 1.97-1.91 (m, 1H), 1.79-1.60 (m, 8H), 1.17-1.07 (m, 5H); ¹³C NMR (100 MHz, d₆-DMSO) δ 171.5, 167.4, 138.6, 133.3, 129.1, 128.8, 128.2, 127.4, 77.1, 71.9, 66.5, 65.3, 57.8, 54.9, 52.4, 52.0, 34.0, 29.6, 27.7, 27.0, 25.6, 25.5, 25.48; HRMS m/z 433.2702 [calcd for C₂₄H₃₆N₂O₅, 433.2702].

[00182] To a solution of the amine 1f (from Step 1 D), 3-hydroxy phenylacetic acid (1.90 g, 12.5 mmol), HOOBt (2.10 g, 12.9 mmol) and EDCl (2.85 g, 14.9 mmol) in anhydrous DMF (250 mL) and CH₂Cl₂ (100 mL) at −20° C., was added NMM (4.20 mL, 38.2 mmol). After being stirred at this temperature for 30 min, the reaction mixture was kept in a freezer overnight (18 h). It was then stirred in air and allowed to warm to room temperature in 1 h. EtOAc (500 mL), brine (100 mL) and 5% H₃PO₄ (100 mL) were added. The separated organic solution was washed with 5% H₃PO₄ (100 mL), saturated aqueous sodium bicarbonate solution (2×150 mL), water (150 mL), and brine (150 mL), dried with magnesium sulfate, filtered and concentrated in vacuo. Flash chromatography (10 to 20% EtOAc-CH₂Cl₂) afforded 1g (6.30 g, 11.1 mmol, 90% (2 steps)) as a white solid. ¹H NMR (400 MHz, d₆-DMSO) δ 9.26 (s, 1H), 8.19 (d, J =8.5 Hz, 1H), 7.36-7.25 (m, 5H), 7.05-7.01 (m, 1H), 6.66-6.64 (m, 1H), 6.60-6.57 (m, 1H), 4.46-4.39 (m, 2H), 4.34 (t, J=8.3 Hz, 1H), 4.29-4.25 (m, 1H), 4.09-4.08 (m, 1H), 3.91 (d, J=11.0 Hz, 1H), 3.66-3.58 (m, 1H), 3.61 (s, 3H), 3.50-3.39 (m, 5H), 3.30 (d, J=13.7 Hz, 1H), 2.24-2.18 (m, 1H), 1.95-1.89 (m, 1H), 1.74-1.57 (m, 8H), 1.18-0.89 (m, 5H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.0, 170.3, 170.0, 157.1, 138.6, 137.6, 128.9, 128.2, 127.4, 127.3, 119.6, 116.1, 113.2, 76.9, 71.8, 66.6, 65.2, 57.4, 54.7, 51.9, 51.8, 41.8, 34.4, 29.6, 28.5, 28.0, 25.5, 25.5; HRMS m/z 567.3073 [calcd for C₃₂H₄₂N₂O₇, 567.3070].

[00183] To a solution of the benzyl ether 1g (6.23 g, 11.0 mmol) in ethanol (200 mL) under nitrogen at room temperature was added 10% Pd-C (1.5 g) cautiously. The resulting suspension was vigorously stirred at room temperature under hydrogen for 23 h. After careful filtration, the solution was concentrated in vacuo. Flash chromatography (2 to 5% MeOH—CH₂Cl₂) afforded 1 h (4.54 g, 9.52 mmol, 87%) as a colorless oil. ¹H NMR (400 MHz, d₆-DMSO) δ 9.26 (s, 1H), 8.22 (d, J=8.6 Hz, 1H), 7.06-7.02 (m, 1H), 6.66-6.58 (m, 3H), 4.42-4.40 (m, 1H), 4.35-4.31 (s, 1H), 4.27 (t, J=8.3 Hz, 1H), 4.10-4.09 (m, 1H), 3.92 (d, J=11.2 Hz, 1H), 3.64 (dd, J=11.2, 4.3 Hz, 1H), 3.61 (s, 3H), 3.59-3.43 (m, 5H), 3.40-3.38 (m,1H), 2.26-2.21 (m, 1H), 1.97-1.90 (m, 1H), 1.74-1.55 (m, 8H), 1.18-0.89 (m, 5H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.0, 170.3, 170.1, 157.1, 137.6, 129.0, 119.6, 116.0, 113.3, 76.9, 65.2, 57.6, 57.4, 54.8, 51.9, 51.8, 41.7, 34.4, 32.6, 28.5, 28.0, 25.9, 25.52, 25.49; HRMS m/z 477.2606 [calcd for C₂₅H₃₆N₂O₇, 477.2601].

[00184] A solution of the phenol alcohol 1h (4.50 g, 9.43 mmol) and ADDP (6.60 g, 26.2 mmol) in anhydrous CH₂Cl₂ was bubbled with argon through a frit glass bubbler for 20 min. To this solution at 0° C. was added triphenylphosphine (4.10 g, 16.3 mmol). After stirring at 0° C. for 20 min, a second portion of triphenylphosphine (3.40 g, 13.5 mmol) was added. The solution was then warmed to room temperature and stirred overnight (24 h) under nitrogen until TLC indicated the complete consumption of the starting material. After removal of solvent in vacuo, the residue was partially purified by flash chromatography (1 to 2% MeOH in CH₂Cl₂) to afford a mixture of the macrocycle 1i and triphenylphosphine oxide. ¹H NMR (400 MHz, d₆-DMSO) δ 8.47 (d, J=9.7 Hz, 1H), 7.17-7.13 (m, 1H), 6.79 (s, 1H), 6.73 (d, J=1.8 Hz, 1H), 6.71 (d, J=1.8 Hz, 1H), 4.50-4.45 (m, 1H), 4.24 (dd, J=10.3, 7.6 Hz, 1H), 4.17-4.06 (m, 4H), 3.68 (d, J=15.1 Hz, 1H), 3.63 (s, 3H), 3.60-3.51 (m, 2H), 3.37 (d, J=15.1 Hz, 1H), 3.35-3.27 (m, 1H), 2.51-2.43 (m, 1H), 1.85-1.47 (m, 9H), 1.22-1.12 (m, 3H), 0.97-0.88 (m, 2H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.0, 170.0, 169.8, 158.4, 138.1, 129.1, 121.8, 115.4, 112.2, 77.0, 64.9, 63.6, 57.0, 54.3, 53.4, 51.8, 41.3, 33.2, 28.9, 28.5, 28.2, 26.0, 25.2; HRMS m/z 459.2495 [calcd for C₂₅H₃₄N₂O₆, 459.2495].

[00185] An aqueous lithium hydroxide solution (0.45 g in 30 mL H₂O ) was added to a 0° C. solution of the methyl ester 1i in THF (30 mL) and methanol (30 mL). The mixture was stirred in an ice bath and warmed to room temperature along with it in 4 h. The progress of the reaction was monitored by TLC. After the volatiles were removed in vacuo, EtOAc (150 mL) and water (30 mL) were added and the two layers separated. The aqueous solution was extracted again with CH₂Cl₂ (150 mL), after which it was acidified to pH=1. EtOAc (200 mL) was then added and the aqueous solution was saturated with solid sodium chloride. After separation of the layers, the aqueous layer was extracted with EtOAc (2×150 mL). Organic solutions were combined, dried with magnesium sulfate, filtered and concentrated in vacuo to afford 1j (1.45 g, 3.26 mmol, 35%, 2 steps) as a pale yellow foam. ¹H NMR (400 MHz, d₆-DMSO) δ 12.32 (bs, 1H), 8.45 (d, J=9.5 Hz, 1H), 7.17-7.13 (m, 1H), 6.73-6.70 (m, 1H), 6.79 (s, 1H), 6.73-6.70 (m, 2H), 4.47 (t, J=9.7 Hz, 1H), 4.17-4.00 (m, 5H), 3.68 (d, J=15.1 Hz, 1H), 3.58-3.45 (m, 2H), 3.39-3.21 (m, 2H), 2.47-2.41 (dd, J=13.4, 7.6 Hz, 1H), 1.85-1.56 (m, 9H), 1.19-1.11 (m, 3H), 0.93-0.87 (m, 2H); ¹³C NMR (100 MHz, d₆-DMSO) δ 173.2, 170.2, 170.0, 158.4, 138.1, 129.3, 122.0, 115.5, 112.2, 77.3, 65.1, 63.8, 57.3, 54.2, 53.7, 41.5, 33.6, 29.0, 28.6, 28.4, 26.1, 25.4; HRMS m/z 445.2335 [calcd for C₂₄H₃₂N₂O₆, 445.2339].

[00186] To a solution of the acid 1j (0.59 g, 1.33 mmol), amine A (H₂N-NVa-CH(OH)-CO-Gly-Phg-NMe₂, 0.55 g, 1.37 mmol), HOOBt (250 mg, 1.53 mmol) and EDCl (315 mg, 1.64 mmol) in anhydrous DMF (50 mL) and CH₂Cl₂ (50 mL) at −20° C. was added NMM (0.50 mL, 4.55 mmol). After stirred at this temperature for 30 min, the reaction mixture was kept in a freezer for 40 h. Then EtOAc (200 mL), brine (50 mL) and 5% H₃PO₄ (50 mL) were added. The separated organic solution was washed, successively, with 5% H₃PO₄ (80 mL), saturated aqueous sodium bicarbonate solution (2×80 mL), water (80 mL), and brine (80 mL), dried with magnesium sulfate, filtered and concentrated in vacuo. Flash chromatography (4 to 7.5% MeOH—CH₂Cl₂) afforded 1k as a mixture of four diastereomers (0.59 g, 0.75 mmol, 56%) as a white solid. ¹ H NMR (400 MHz, d₆-DMSO) δ 8.54-8.35 (m, 2H), 7.95-6.98 (m, 8H), 6.79-6.77 (m, 1H), 6.72-6.70 (m, 2H), 5.96-5.73 (m, 2H), 4.53-4.45 (m, 1H), 4.35-3.61 (m, 11H), 3.54-3.41 (m, 1H), 3.40-3.22 (m, 1H), 2.93-2.92 (m, 3H), 2.84 (s, 3H), 2.42-2.17 (m, 1H), 1.87-1.55 (m, 10H), 1.49-1.06 (m, 7H), 0.98-0.75 (m, 5H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.1, 171.9, 171.8, 170.8, 170.7, 170.4, 170.4, 170.3, 170.0, 169.8, 169.7, 169.6, 169.2, 169.2, 167.9, 167.8. 167.8, 158.4, 158.3, 138.2, 138.15, 138.11, 138.07, 137.6, 137.50, 137.48, 129.1, 128.5, 128.3, 127.8, 127.7, 121.7, 121.6, 115.4, 115.3, 112.09, 112.07, 112.0, 111.9, 76.9, 76.8, 73.3, 72.1, 71.9, 64.9, 64.8, 63.2, 58.7, 58.5, 57.9, 57.8, 54.6, 54.5, 54.48, 53.8, 53.78, 53.7, 53.66, 53.0, 52.9, 51.0, 50.8, 50.7, 41.6, 41.5, 41.4, 41.3, 36.6, 35.3, 33.9, 33.86, 33.5, 32.9, 32.1, 29.9, 29.0, 28.9, 28.5, 28.4, 28.3, 26.0, 25.9, 25.3, 25.25, 25.2, 18.6, 18.56, 18.5, 13.8, 13.7; HRMS m/z 791.4339 [calcd for C₄₂H₅₈N₆O₉, 791.4344, error=1 ppm].

Example 2

Preparation of Compound 2

[00188]

[00189] The desired compound 2a was prepared according to the method of Example 1, Step J, except for substituting the amine B for the amine A. The hydoxy amide was obtained as a mixture of inseparable diastereomers in the form of a white solid in 60% yield.

Example 3

Preparation of the Compound of Formula 3 below

[00191]

Example 4

Preparation of Compounds of Formulas 4A and 4B

[00193]

[00194] The desired product 4a was obtained by the method described for Example 1, Step A. The crude material was carried to the next step as it was.

[00195] The desired product 4b was obtained by the method described for Example 1, Step B. The material was purified by flash column chromatography using 80/20 to 75/25 hexanes/ethyl acetate to provide 4b in 50% yield as a colorless oil.

[00196] The desired compound 4c was prepared by the protocol described for Example 1, Step C. The material was carried forward as it was.

[00197] The desired product 4d was obtained by the method described in Example 1, Step D. The material after work-up was sufficiently pure to be carried to the next step. HRMS (FAB) Calcd for C₃₀H₄₇N₂O₇: 547.3383 (M+H)⁺. Found: 547.3372.

[00198] The desired product 4e was obtained by the method described in Example 1, Step E. The crude material was carried to the next step as was.

[00199] The desired product 4f was obtained by the method described in Example 1, Step F. The material was purified by flash column chromatography using 80/20 to 60/40 dichloromethane/ethyl acetate to provide 4f in 85% yield. HRMS (FAB) Calcd for C₃₃H₄₅N₂O₇: 581.3227 (M+H)⁺. Found: 581.3222.

[00200] The desired product 4g was obtained by the method described in Example 1, Step G. The crude material was carried to the next step as was. HRMS (FAB) Calcd for C₂₆H₃₉N₂O₇: 491.2757 (M+H)⁺. Found: 491.2761.

[00201] The desired product 4h was obtained by the method described in Example 1, Step H. Purification by column chromatography using 99/1 dichloromethane/methanol afforded 4h along with triphenylphosphine oxide. This mixture was taken to the next step.

[00202] The desired product was obtained by the method described in Example 1 Step I. Yield of 4i (for 2 steps)=24%. ¹H NMR (DMSO-d₆) δ 0.90-0.95 (m, 2H), 1.10-1.16 (m, 3H), 1.51-1.79 (m, 11H), 2.43 (dd, 1H), 3.29-3.32 m, 2H), 3.50-3.54 (m,1H), 3.62-3.68 (m, 2H), 3.91-3.99 (m, 3H), 4.04-4.08 (m, 2H), 4.46 (t, 1H), 6.67-6.72 (m, 3H), 7.13 (app. t, 1H), 8.36 (d, 1H), 12.40 (br. s, 1H); ¹³C NMR (DMSO-d₆) δ 25.26, 25.31, 25.97, 26.62, 28.42, 33.28, 39.75, 41.49, 53.50, 54.28, 57.45, 67.57, 67.98, 77.25, 111.07, 115.23, 121.48, 129.11, 137.99, 158.33, 170.07, 172.92; HRMS (FAB) Calcd for C₂₅H₃₅N₂O₆: 459.2495 (M+H)⁺. Found: 459.2494.

[00203] The expected product 4j was synthesized as described earlier for the Example 1 Step J. The material after work-up was of sufficient purity to be carried forward to the next step. HRMS (FAB) Calcd for C₄₃H₆₁ N₆O₉: 805.4500 (M+H)⁺. Found: 805.4492.

Example 5

Preparation of Compound 5

[00205]

[00206] The expected product 5a was synthesized as described earlier for the Example 2, Step A. The material after work-up was of sufficient purity to be carried forward to the next step. HRMS (FAB) Calcd for C₄₅H₆₄N₅O₁₀: 834.4653 (M+H)⁺. Found: 834.4648.

Example 6

Preparation of Compound 6

[00208]
The expected product 6 was synthesized as described earlier for the Example 3, Step A in quantitative yield. HRMS (FAB) Calcd for C₄₁ H₅₄N₅O₁₀: 776.3871 (M+H)⁺. Found: 776.3865.

Example 7

Preparation of compounds 7A and 7B

[00210]

[00211] The desired compound 7a was prepared from 1c according to the procedure of Example 1, Step A. The crude product was used in Step B without further purification.

[00212] The desired compound 7b was prepared from 7a according to the procedure of Example 1, Step B.

[00213] The desired compound 7c was prepared from 7b according to the procedure of Example 1, Step C. The product was used in Step D without further purification.

[00214] The desired compound 7d was prepared from 7c according to the procedure of Example 1, Step D.

[00215] The desired compound 7e was prepared from 7d according to the procedure of Example 1, Step E. The product was used in Step F without further purification.

[00216] The desired compound 7f was prepared from 7e according to the procedure of Example 1, Step F.

[00217] The desired compound 7g was prepared from 7f according to the procedure of Example 1, Step F.

[00218] A solution of the phenol alcohol 7g (830 mg, 1.79 mmol) and ADDP (1.36 g, 5.39 mmol) in anhydrous CH₂Cl₂ (200 mL) was bubbled with Argon through a frit glass bubbler for 20 min. To this solution at 0° C. was added triphenylphosphine (1.41 g, 5.38 mmol). After stirring at 0° C. for 20 min, the solution was warmed to room temperature and stirred overnight (20 h) under nitrogen. After removal of solvent in vacuo, the residue was purified by flash chromatography (1 to 3% MeOH in CH₂Cl₂) to afford a mixture of the desired product 7 h and triphenylphosphine oxide, which was used in Step I without further purification.

[00219] The desired compound 7i was prepared from 7h in 36% yield (2 steps) according to the procedure of Example 1, Step I.

[00220] The desired compound 7j was prepared from 7i and A in 56% yield according to the procedure of Example 1, Step J.

Example 8

Preparation of Compound of Formula 8

[00222]

[00223] The desired compound 8a was prepared according to the method of Example 1, Step J, except substituting amine B for amine A. The product was obtained as a mixture of inseparable diastereomers in the form of a white solid in 57% yield.

Example 9

Preparation of Compound of Formula 9

[00225]

Example 10

preparation of Compounds of Formula 10A and 10B

[00227]

[00228] To a 0° C. solution of 10a (10 g, 41 mmol) in dichloromethane (60 mL) was added triethylamine (28.68 mL, 204 mmol) slowly. 4-bromobenzenesulfonyl chloride (20.91 g, 82 mmol) and DMAP (few crystals) were then added and the temperature was maintained at 0° C. for 30 min. The reaction mixture was left standing in the refrigerator (˜5° C.) overnight followed by slow warming to ambient temperature over two hours. At this time TLC analysis revealed complete consumption of the starting material. The reaction mixture was diluted with dichloromethane, washed with saturated sodium bicarbonate solution, and 10% aqueous citric acid solution. The organic layer was dried (Na₂SO₄) and concentrated. The crude mixture was purified by flash column chromatography using 100/0 to 95/5 dichloromethane/ethyl acetate to provide 18.4 g (97% yield) of the brosylate 10b as a white solid; ¹H NMR (mixture of rotamers, CDCl₃) δ 1.41 and 1.45 (2s, 9H), 2.40-2.50 (m, 2H), 3.59-3.69 (m, 5H), 4.33-4.37 and 4.46 (2 dd, 1H), 5.11 (m, 1H), 7.72-7.74 (m, 4H); ¹³C NMR (mixture of rotamers, CDCl₃) δ 28.18, 28.27, 36.01, 36.98, 51.59, 52.03, 52.20, 52.35, 56.95, 57.22, 57.28, 78.35, 79.53, 80.66, 129.10, 129.26, 132.66, 135.66, 135.81, 153.25, 153.64, 171.45, 171.78; HRMS (FAB) Calcd for C₁₇H₂₃NO₇SBr: 464.0379 (M+H)⁺. Found: 464.0375.

[00229] To a suspension of sodium hydride (60% dispersion in mineral oil, 187 mg, 4.68 mmol) in DMF at 0° C. was added 3-mercaptopropanol (0.42 mL, 4.85 mmol) under argon atmosphere. The mixture was stirred for 30 min while maintaining the temperature. A solution of brosylate 101b (1.5 g, 3.23 mmol) in DMF (total volume=10 mL) was added slowly and the mixture was warmed to ambient temperature over 2 hrs. The reaction was quenched by pouring into cold 10% citric acid solution. The aqueous layer was extracted with ethyl acetate, the organic layer was dried (Na₂SO₄) and concentrated. The crude material was purified by flash column chromatography using 85/15 dichloromethane/ethyl acetate to provide 800 mg (78% yield) of the sulfide 10c as an oil; ¹H NMR (mixture of rotamers, CDCl₃) δ 1.41 and 1.47 (2s, 9H), 1.83-1.89 (m, 2H), 2.13-2.34 (m, 2H), 2.69 (t, 2H), 3.23-3.49 (m, 2H), 3.73-3.78 (m, 5H), 3.86-3.95 (m, 1H), 4.33-4.37 and 4.42-4.46 (2dd, 1H); ¹³C NMR (mixture of rotamers, CDCl₃) δ 28.21, 28.30, 32.15, 32.23, 36.65, 37.27, 40.45, 40.89, 52.16, 52.35, 52.50, 52.84, 58.32, 58.55, 61.22, 61.41, 80.35, 153.49, 153.99, 173.05, 173.23; HRMS (FAB) Calcd for C₁₄H₂₆NO₅S: 320.1532 (M+H)⁺. Found: 320.1528.

[00230] The desired compound 10d was prepared by the protocol described for Example 1, Step C. Reaction conditions were 0° C., 1 hr. The material was carried to the next step as it was.

[00231] The desired compound 10e was prepared by the method described for Example 1, Step D. The coupling reaction was carried out at −8° C. for 2 days. After workup the product 10e was sufficiently pure by TLC and was obtained in 80% yield; HRMS (FAB) Calcd for C₂₂H₃₉N₂O₆S: 459.2529 (M+H)⁺. Found: 459.2523.

[00232] The desired compound 10f was prepared by the protocol described for Example 1, Step E. The material was carried forward as it was.

[00233] The desired compound 10g was prepared by the procedure described for Example 1, Step F. The crude product was purified by flash column chromatography using 98/2 of dichloromethane/methanol to provide 10 g in 40% yield as a white solid; ¹H NMR (mixture of rotamers, CDCl₃) δ 0.90-1.26 (m), 1.66-1.88 (m), 2.22-2.31 (m, 2H), 2.73 (t, 2H), 3.47 (s), 3.5-3.55 (m), 3.65-3.75 (m), 3.88-3.94 (dd, 1H), 4.07-4.12 (dd, 1H), 4.53 (t, 1H), 4.62 (t, 1H), 6.73-6.80 (m, 4H), 7.17 (t, 1H); ¹³C NMR (mixture of rotamers, CDCl₃) δ 25.80, 25.89, 26.14, 27.71, 28.55, 29.22, 31.88, 35.46, 40.58, 42.44, 43.16, 52.32, 52.90, 55.49, 58.46, 60.30, 114.59, 116.27, 121.01, 130.02, 135.90, 156.73, 171.25, 171.87, 171.96; HRMS (FAB) Calcd for C₂₅H₃₇N₂O₆S: 493.2372 (M+H)⁺. Found: 493.2364.

[00234] The desired compound 10h was prepared by the protocol described for Example 1, Step G. The crude product was suspended in 80/20 ethyl acetate/hexane, and the solid material was filtered off. The filtrate was concentrated and purified by flash column chromatography using 80/20 hexane/acetone to yield 22% of 10h as a solid. ¹H NMR (CDCl₃) δ 0.98-1.30 (m), 1.64-1.90 (m), 2.06-2.14 (m, 1H), 2.16-2.21 (dd, 2H), 2.62-2.70 (m, 2H), 3.38-3.46 (m, 2H), 3.60-3.66 (m, 3H), 3.71 (s, 3H), 3.88-3.94 (dd, 1H), 4.07-4.15 (m, 1H), 4.22-4.29 (m,1H), 4.48 (t,1H), 4.60 (t,1H), 5.97 (br t,1H), 6.76-6.81 (m, 2H), 6.99 (br s,1H), 7.20 (dd, 1H); HRMS (FAB) Calcd for C₂₅H₃₅N₂O₅S: 475.2267 (M+H)⁺. Found: 475.2260.

[00235] The advanced intermediate 10i was synthesized as described for Example 1, Step H, in quantitative yield as a white solid; ¹H NMR (DMSO-d₆) δ 0.88-0.96 (m, 2H), 1.10-1.14 (m, 3H), 1.59-1.76 (m, 7H), 1.88-1.94 (m, 1H), 2.09 (app. t, 1H), 2.61 (dd,1H), 3.32 (app. d,1H), 3.40-3.45 (m, 2H), 3.61 (app. d, 1H), 3.83 (q, 1H), 4.13 (app. t, 1H), 4.19 (t, J=7.32Hz, 1H), 4.40 (t, J=9.52Hz, 1H), 6.76-6.79 (m, 2H), 6.89 (s, 1H), 7.16 (app. t, 1H), 8.39 (d, 1H), 12.5 (br. s, 1H); ¹³C NMR (DMSO-d₆) δ 25.33, 25.41, 26.01, 26.44, 28.09, 28.62, 29.24, 34.90, 39.50, 41.40, 42.30, 53.18, 54.44, 58.06, 66.94, 114.88, 115.25, 122.28, 129.20, 137.84, 157.90, 169.25, 170.29, 172.59; HRMS (FAB) Calcd for C₂₄H₃₃N₂O₅S: 461.2110 (M+H)⁺. Found: 461.2104.

[00236] The desired compound 10j was prepared as described earlier for Example 1, Step I, in quantitative yield as a pale yellow solid. The material obtained after workup was sufficiently pure by TLC for further manipulations; HRMS (FAB) Calcd for C₄₂H₅₉N₆O₈S: 807.4115 (M+H)⁺. Found: 807.4103.

Example 11

Preparation of Compound of Formula 11

[00238]

[00239] To a cold (0° C.) solution of 10h (200 mg, 0.42 mmol) in dichloromethane (10 mL) was added MCPBA (60%, 364 mg, 1.26 mmol). The reaction mixture was slowly warmed to ambient temperature over 16 hrs. The reaction mixture was diluted with dichloromethane and washed with saturated sodium bicarbonate and sodium bisulfite solution. The organic layer was dried (Na₂SO₄), and concentrated. Purification by flash column chromatography using 98/2 dichloromethane/methanol provided 11a (138 mg, 65% yield). HRMS (FAB) Calcd for C₂₅H₃₅N₂O₇S: 507.2165 (M+H)⁺. Found: 507.2158.

[00240] The expected product 11b was synthesized as described for Example 1, Step I in 90% yield as a white solid; HRMS (FAB) Calcd for C₂₄H₃₃N₂O₇S: 493.2008 (M+H)⁺. Found: 493.2012.

[00241] The desired compound 11c was prepared as described earlier for Example 1, Step J, in quantitative yield. The material obtained after workup was sufficiently pure by TLC for further manipulations. HRMS (FAB) Calcd for C₄₂H₅₉N₆O₁₀S: 839.4013 (M+H)⁺. Found: 839.4019.

Example 12

Preparation of Compounds of Formulas 12A and 12B

[00243]

[00244] The desired product 12a was obtained by the method described for Example 1, Step D using N-boc-tert-butylglycine as the coupling partner. The material was purified by flash column chromatography using 90/10 dichloromethane/ethyl acetate to provide 12a in 73% yield. ¹³C NMR (mixture of rotamers, CDCl₃) δ 26.20, 28.31, 29.07, 30.06, 34.94, 35.86, 37.06, 51.21, 52.16, 52.84, 57.78, 58.33, 65.95, 66.92, 72.97, 75.48, 79.45, 127.55, 127.66, 128.35, 138.45, 155.62, 165.06, 171.13, 172.54; HRMS (FAB) Calcd for C₂₇H₄₃N₂O₇: 507.3070 (M+H)⁺. Found: 507.3077.

[00245] The desired compound 12b was prepared by the protocol described for Example 1, Step E. The material was carried forward to the next step.

[00246] The desired product 12c was obtained by the procedure described for Example 1, Step F. The material was purified by flash column chromatography using 99/1 dichloromethane/methanol to yield 12c in 91%. ¹³C NMR (CDCl₃) δ 26.24, 29.93, 34.95, 35.96, 43.48, 52.18, 53.09, 57.06, 58.06, 66.10, 66.92, 72.93, 77.43, 114.59, 116.14, 120.87, 127.58, 127.64, 127.74, 128.37, 130.02, 135.95, 138.39, 156.90, 170.65, 171.06, 172.38; HRMS (FAB) Calcd for C₃₀H₄₁N₂O₇: 541.2914 (M+H)⁺. Found: 541.2921.

[00247] The desired product 12d was obtained by the procedure described for Example 1, Step G. The product obtained after filtering off the catalyst was pure enough for subsequent manipulations. ¹³C NMR (CDCl₃) δ 26.27, 32.09, 35.44, 35.67, 43.19, 52.21, 52.74, 57.60, 58.21, 58.75, 65.78, 77.74, 114.74, 116.02, 120.68, 130.07, 135.66, 157.11, 170.59, 172.05, 172.51; HRMS (FAB) Calcd for C₂₃H₃₅N₂O₇: 451.2444 (M+H)⁺. Found: 451.2436.

[00248] The desired product 12e was obtained by the procedure described for Example 1, Step H. The crude material was suspended in ethyl acetate/hexane (approx. 1/1) and the undissolved solid material was filtered off. Repeated this process once again, the filtrate was concentrated and applied on the column as a dichloromethane solution. The column was eluted with 75/25 hexane/acetone to yield 29% of 12e. HRMS (FAB) Calcd for C₂₃H₃₃N₂O₆: 433.2339 (M+H)⁺. Found: 433.2339.

[00249] The advanced intermediate 12f was synthesized as described for Example 1, Step I, in quantitative yield; ¹H NMR (DMSO-d₆) δ 0.96 (s, 9H), 1.66-1.70 (m, 1H), 1.75-1.82 (m, 2H), 2.43 (dd,1H), 3.32-3.36 (m, 2H), 3.48-3.52 (m, 1H), 3.55 (dd, 1H), 3.84 (app. d, 1H), 3.99 (app. d, 1H), 4.06-4.10 (m, 3H), 4.16 (dd, 1H), 4.69 (d, 1H), 6.70-6.72 (m, 3H), 7.15 (app. t, 1H), 8.42 (d, 1H), 12.43 (br. s, 1H), ¹³C NMR (DMSO-d₆) δ 26.25, 28.54, 33.31, 34.97, 41.22, 53.96, 56.11, 56.97, 63.36, 64.96, 76.84, 111.94, 115.25, 121.73, 129.13, 138.36, 158.27, 169.85, 170.15, 173.04; HRMS (FAB) Calcd for C₂₂H₃₁N₂O₆: 419.2182 (M+H)⁺. Found: 419.2180.

[00250] The expected product 12g was synthesized as described earlier for the Example 1, Step J. The material after work-up was of sufficient purity to be carried forward to the next step. HRMS (FAB) Calcd for C₄₀H₅₇N₆O₉: 765.4187 (M+H)⁺. Found: 765.4175.

Example 13

Preparation of Compounds of Formulas 13A and 13B

[00252]

[00253] The expected product 13a was synthesized as described earlier for the Example 1, Step J. The material after work-up was of sufficient purity to be carried forward to the next step. HRMS (FAB) Calcd for C₄₁H₅₉N₆O₉: 779.4344 (M+H)⁺. Found: 779.4350.

Example 14

Preparation of Compound 14

[00255]

[00256] A solution of vinyl benzoic acid 14a (10 g, 68 mmol) in dry benzene (150 mL) was treated with ditert-butylacetal of DMF (69 g, 340 mmol, 5.0 equiv.) and heated at reflux for 4 h. The reaction mixture was concentrated in vacuo and diluted with aq. NaOH (1M, 300 mL). The reaction mixture was extracted in diethyl ether (3×100 mL). The combined organic layer was extracted with aq. NaOH (1M, 100 mL), H₂O (2×100 mL), brine (1×100 mL) dried (Na₂SO₄), filtered and concentrated in vacuo. The residue was distilled under reduced pressure to yield 14b 9.2 g (66.2%) of a colorless oil.

[00257] A solution of tert-butyl carbamate (5.96 g, 50.9 mmol) in 1-PrOH (68 mL) was treated with aq. NaOH (128 mL, 0.41 M ) and tert-butyl hypochlorite (5.5g, 50.9 mmol). The reaction mixture was cooled to 0° C. and (DHQ)₂Phal (780 mg, 1.00 mmol) in 1-PrOH (64 mL) was added. A solution of tert-butyl-4-vinylbenzoate 14b in 1-PrOH (119 mL) was added followed by K₂OsO₄•H₂O (248 mg, 0.7 mmol) and the reaction mixture was stirred at 0° C. for 4-5 h. The reaction turns green and all the starting material disappears with a new product formed. The reaction mixture was concentrated in vacuo and the residue was diluted with H₂O (300 mL) and extracted with EtOAc (3×100 mL). The combined organic layer was extracted with aq. HCl (200 mL), brine(100 mL) dried (Na₂SO₄) filtered conc. in vacuo and purified by chromatography (SiO₂, EtOAc/Hexanes 1:2) to yield 14c as a colorless solid (4.6 g, 82%).¹H NMR (CD₃OD, δ) 7.90 (d, 2H, J=6.0 Hz), 7.40 (d, 2H, J=6.3 Hz), 7.22 (bd, 1H, J=5.7 Hz), 4.69 (bs, 1H), 3.71-3.62 (m, 2H) 1.58 (s, 9H), 1.39 (s, 9 Hz); ¹³C NMR (CD ₃OD, 75 MHz) 169.7, 160.5, 149.8, 134.5, 132.9, 130.4, 84.7, 82.9, 68.7, 60.7, 31.25, 30.9-MS (FAB) 675.2 ([2M+1]⁺, 15), 338 ([M+1]⁺, 15), 282 (65), 225 (50), 165 (100); HRMS calcd for C₁₈H₂₈NO₅ (M+1): 338.1887; found 338.1967.

[00258] A solution of aromatic compound 14c (1.0 g, 2.96 mmol) in CH₃ OH (20 mL) was treated with Rh/C (10% w/w 100 mg) and hydrogenated (60 psi) for 2d. The reaction mixture was filtered through a plug of celite and the residue was concentrated in vacuo to yield 14d. The crude product was purified by chromatography (SiO₂, EtOAc/Hex 2:3) to yield the cis compound 14d (830 mg, 83%) which was further purified by crystallizing from hexanes. ¹H NMR (CD₃OD, δ) 6.31 (d, 1H, J=6.9 Hz), 3.58-3.49 (s, 2H), 3.40 (bd, 1H, J=4.8 Hz), 2.48-2.46 (m, 1H), 2.1-1.98(m, 2H), 1.61-1.2 (m, 7H), 1.45 (s, 9H), 1.42 (s, 9H), ¹³C NMR (CD ₃OD, 75 MHz) 176.2, 158.5, 81.2, 79.8, 63.1, 57.2, 41.8, 38.8, 28.8, 28.3, 27.7, 27.5, 25.9. MS (FAB) 687.2 ([2M+1]⁺, 5), 344 ([M+1]⁺, 20), 232 (40), 188(100),107 (13); HRMS calcd for C₁₈H₃₄NO₅ (M+1): 344.2437; Found: 344.2444. CHN calcd for C₁₈H₃₃NO₅ C=64.07%, H=8.07%,N=4.15% Found C=64.32%, H=8.21%, N=4.32%.

[00259] A solution of amino alcohol 14d (3.3 g, 11.08 mmol) in dry THF (200 mL) was cooled to −78° C. (dry ice/acetone, internal temperature −68° C.) was treated with LDA (44 mL, 2M soln in heptanes, 88 mmol, 8.0 equiv.). The reaction mixture was stirred at −78° C. for 2 h and quenched with CH₃ OH (20 mL). The reaction mixture was treated with aq. HCl (150 mL, 1 M) and extracted with ether (3×100 mL). The combined ether layer was extracted with brine (50 mL) dried (MgSO₄), concentrated in vacuo and purified by crystallization from boiling hexanes. The solid separating out from the mother liquor was predominantly cis stereo isomer, where concentration of the mother liquor gave the pure trans isomer. The above sequence was repeated twice more to obtain 2.7 g of the trans compound and 600 mg of cis/trans mixture. ¹³C NMR (CDCl3, 75 MHz) 175.3, 156.6, 79.8, 63.6, 57.0, 44.1, 38.3, 37.7, 28.9, 28.6, 28.4, 28.1, 26.6, 26.1. MS (electron spray) 344 (M⁺, 50), 288 (50) 232 (90), 188 (100).

[00260] A solution of alcohol 14e (2.6 g, 7.6 mmol) in CH3CN (150 mL) and CCl₄ (150 mL) was treated with H₂O (22 mL), cooled to 0° C., treated with periodic acid (7.05 g, 30.92 mmol, 4.0 equiv.) and RuCl₃•3H₂O (60 mg, 0.3 mmol, 4 mol %). The reaction mixture was stirred at rt. for 3h, and concentrated in vacuo. The residue was diluted with water (150 mL) and extracted with EtOAc (3×100 mL). The combined organic layer was extracted with H₂O (100 mL) and with aq. NaOH (1M, 3×100 mL).The combined aq. layers were acidified with HCl (6M, pH ˜1) and extracted with EtOAc (3×100 mL). The ethyl acetate layers were pooled, extracted with brine (100 mL) dried (Na₂SO₄) filtered concentrated in vacuo to yield acid 14f (1.8 g, 66%) used for further couplings without further purification. MS (FAB) 380.2 ([M+Na]⁺, 30)358 ([M+1]⁺, 5), 302 (20), 258(20), 246 (100), 202 (70), 200 (20)

[00261] A solution of Boc-trans-4-tertbutylcarboxyl-cyclohexylgylcine 14f (1.9 g, 5.3 mmol) in CH₂Cl₂ (30 mL) was treated with proline compound 1d (1.92 g, 5.85 mmol, 1.1 equiv.) and cooled to 0° C. The reaction mixture was treated with Hünigs base (1.51 g, 11.7 mmol, 2.2 equiv., 2.15 mL) followed by the addition of BOP reagent (2.6 g, 5.85 mmol, 1.1 equiv.) The reaction mixture was stirred at rt. for 12 h diluted with aq. HCl (1M, 100 mL) and extracted with EtOAc (3×100 mL). The combined ethyl acetate layers were extracted with aq. NaOH (1M, 100 mL) brine (100 mL) dried (Na₂SO₄), filtered concentrated in vacuo and purified by chromatography (SiO₂, EtOAc/Hexanes 2:3) to yield 14g as colorless foam (1.8 g, 54%); ¹H NMR (CD₃OD, δ, mixture of rotomers) 7.32-7.23 (m, 5H), 6.64 (d, 1H, J=9.0 Hz), 4.47-4.39 (m, 3H), 4.19-4.04 (m, 3H), 3.74 (s, 3H), 3.66-3.56 (m, 4 H), 2.55-2.10 (m, 2H), 1.99-1.00 (m, 12H), 1.42 (s, 9H), 1.40 (s, 9H). ¹³C NMR (CD₃OD, δ, mixture of rotamers), 175.6, 174.6, 172.4, 172.0, 156.4, 138.5, 128.1, 127.5, 127.4, 127.3, 79.8, 79.7, 79.0, 77.5, 72.5, 66.7, 65.4, 58.1, 56.6, 52.1, 51.3, 43.9, 40.4, 39.4, 38.6, 34.6, 29.8, 28.4, 27.9, 27.3, 26.1, 25.4, 24.5; MS (FAB) 633 ([M+1]+, 11), 533 (55), 477 (24), 428 (5), 294 (100), 234 (12) 156 (40), 128 (39); CHN Calcd. for C₃₄H₅₂NO₉ C 64.53% H 8.28% N 4.43%; Found C 64.41% H 8.00% N 4.19%.

[00262] A solution of Boc-trans-4-tertbutylcarboxycyclohexyl glycine 14g (1.8 g) in HCl (4M soln in dioxane, 60 mL) was stirred at rt. for 4-5 h. The reaction was followed by TLC (EtOAc/Hex 3:7) for the disappearance of starting material and the appearance of base line product. The reaction mixture was concentrated in vacuo and the residue was dried in the pump overnight. The solid 14h was used for couplings without further purification.

[00263] A solution of 3-hydroxyphenylacetic acid 14i (501 mg, 3.29 mmol, 1.8 equiv.) and amine hydrochloride 14h (1.79 g, 2.99 mmol) in dry CH₂Cl₂ (30 mL) was treated with Hünigs base (850 mg, 6.59 mmol, 2.20 equiv., 1.2 mL) and BOP reagent (1.5 g, 3.29 mmol, 1.1 equiv.) at 0° C. and stirred at rt. for 24 h. The reaction mixture was concentrated in vacuo and diluted with aq. HCl (1 M, 250 mL). The aq. layer was extracted with EtOAc (3×100 mL). The combined organic layers were extracted with aq. NaOH (1×100 mL), brine (1×100 mL) dried (Na₂SO₄) filtered, concentrated in vacuo and purified by chromatography (EtOAc/Hexanes 1:1) to yield 14j as a colorless solid (710 mg, 36%).

[00264] A solution of the coupled compound 14j (710 mg, 1.1 mmol) in CH3OH (50 mL) was treated with Pearlmans catalyst (10% Pd(OH)₂/C) and hydrogenated (H₂, 40 psi) for 12 h. The Pd/C was filtered off through a plug of celite and filtrate was concentrate and used for the next cyclization without further purification R_f 0.12 (acetone/Hexanes 3:7); ¹H NMR (CD₃OD, δ, mixture of rotomers) 8.25 (bs, 1H), 7.01 (bt, 1H, J=7.2 Hz), 6.72 (bs, 1H), 6.65 (d, 2H, J=7.8 Hz), 4.79-4.70 (bs, 3H), 4.55-4.41 (bs, 2H), 4.20-4.12 (bs, 2H), 3.77 (s, 3H), 3.66-3.44 (bm, 6H) 2.43-1.04 (bm, 14H) 1.40 (s, 9H); ¹³C NMR (CD₃OD, δ, mixture of rotomers): 175.5, 174.6, 172.6, 172.4, 171.3, 161.1, 157.3, 136.8, 136.7, 129.3, 120.0, 115.7, 113.6, 100.0, 94.8, 79.9, 79.8, 77.5, 65.1, 58.5, 58.2, 55.6, 55.5, 54.3, 5.2, 51.5, 43.9, 42.1, 39.4, 35.0, 32.4, 28.6,28.4,27.5, 27.2, 27.1, 26.1, 25.2, 25.0; MS (FAB): 577 ([M+1]⁺, 70), 521(10), 443 (10), 387 (10), 374 (10), 318 (15), 290 (100), 248 (30); HRMS calcd. for C₃₀H₄₅N₂O₉ (M+H)⁺: 577.3125; Found 577.3133.

[00265] A solution of alcohol 14k (600 mg, 1.05 mmol) and ADDP (787 mg, 3.12 mmol, 3.0 equiv.) in CH₂Cl₂ (30 mL) was treated with Ph₃P (818 mg, 3.12 mmol, 3.0 equiv.) under positive pressure of dry N₂ at 0° C. The reaction mixture was stirred at rt. for 24 h, concentrated in vacuo and the residue was purified by chromatography (SiO₂, acetone/Hexanes 2:3) to yield a colorless solid 14l (120 mg, 22%) of cyclic product; R_f 0.73 (acetone/Hexanes 1:1); ¹H NMR (CD₃OD, δ) 7.13 (t, 1H J=7.8 Hz), 6.76 (s, 1H), 6.71 (t, 2H, J=8.1 Hz), 4.58 (d, 1H, J=9.9 Hz), 4.37 (dd, 1H, J=7.8, 2.7 Hz), 4.23-4.11 (m, 4H), 3.74-3.61 (m, 2H), 3.66 (s, 3H), 3.59-3.40 (m, 2H), 2.54-2.41 (m, 1H), 2.19-2.10 (m, 1H), 1.98-1.42 (m, 10H), 1.43 (s, 9H); ¹³C NMR (CD₃OD, δ) 175.6, 172.4, 172.1, 170.8, 159.0, 137.0, 129.1, 121.7, 115.7, 112.2, 94.8, 79.8, 77.4, 65.2, 64.0, 57.6, 55.2, 53.8, 51.3, 44.0, 41.5, 39.1, 33.3, 28.7, 28.1, 26.9; MS (electron spray) 559 ([M+1]⁺, 100) 327 (10), 189 (20); HRMS calcd. for C₃₀H₄₃N₂O₈ (M+1)⁺: 559.3019; Found: 559.3025; CHN calcd for C₃₀H₄₂N₂O₈.0.5H₂O C=63.47%, H=7.63%, N=4.93%; Found C=63.57%, H=7.46%, N=4.93%.

[00266] A solution of methyl ester 14I (120 mg, 0.22 mmol) in THF (5.0 mL) and H₂O (1.0 mL) was treated with LiOH (20 mg, 0.5 mmol, 2.0 equiv.). The reaction mixture was stirred at rt. for 3 h and CH₃OH (1.0 mL) was added and stirred for an additional 1 h. The reaction mixture was stirred with HCl (4.0 M in Dioxane, 1 mL) and concentrated in vacuo and the water was lyophilized to yield a colorless solid 14m which was used for next coupling.

[00267] A solution of carboxylic acid 14m (110 mg, 0.21 mmol) in DMF (3.0 mL) and CH₂Cl₂ (5.0 mL) was treated with Hünigs base (109 mg, 0.84 mmol, 4.0 equiv. 155.0 μL) and HOOBt (52 mg, 0.315 mmol, 1.5 equiv.). The reaction mixture was cooled to 0° C. and treated with EDCl (61 mg, 0.31 mmol, 1.5 equiv.) The reaction mixture was stirred at 0° C. and treated with amine hydrochloride A after 30 min. The reaction mixture was stirred at rt. for 24 h and concentrated in vacuo to remove DMF and CH₂Cl₂. The residue was diluted with aq. HCl (100 mL) and extracted with CH₂Cl₂ (3×75 mL) The combined organic layers were extracted with aq. NaOH (1M, 3×50 mL), brine (100 mL) and concentrated in vacuo. The residue 14n (79 mg) was oxidized without further purification.

Example 15

Preparation of Compound 15

[00269]

Example 16

Preparation of Compound 16

[00271]

[00272] The desired compound 16a was prepared from 14d in 70% yield according to the procedure of Example 14, Step F. It was used for the couplings without further purification's; MS (FAB): 380.2 ([M+Na]⁺, 30) 358 ([M+1]⁺5), 302 (20), 258(20), 246 (100), 202 (70), 200 (20); HRMS calcd. for C₁₈H₃₂NO₆ (M+1)⁺: 358.2230; Found: 358.2237.

[00273] The desired compound 16b was prepared from 16a in 41% yield according to the procedure of Example 14, Step G; [α]_D −52.7 (c 0.3 CHCl₃, 25); ¹H NMR (CDCl₃, δ) 7.35-7.21 (m, 5H), 6.63 (d, 1H, J=9.3 Hz), 4.46 (d, 2H, J=4.3 Hz), 4.41 (t, 1H, J=9.3 Hz), 4.38-4.07 (m, 3H), 3.68 (s, 3H), 3.66-3.43 (m, 5H), 2.45 (p, 1H, J=4.2 Hz), 2.32 (dd, 1H, J=7.8, 5.7 Hz), 2.02-1.90 (m, 3H), 1.90-1.56 (m, 3H), 1.56-1.24 (m, 24H); ¹³C NMR (CD₃OD, δ)174.7, 172.4, 172.0, 156.4, 138.4, 128.0, 127.5, 127.4, 79.8, 79.0, 77.5, 72.5, 66.6, 65.3, 58.0, 55.3, 52.1, 51.3, 40.4, 38.6, 34.7, 29.7, 27.4, 27.03, 26.1, 24.5. MS (FAB) 633.2 [(M+1)⁺, 100]; HRMS calcd for C₃₄H₅₃N₂O₉ (M+1)⁺: 633.3751; Found 633.3759.

[00274] The desired compound 16c was prepared from 16b according to the procedure of Example 14, Step H. The product was used without further purification.

[00275] The desired compound 16d was prepared from 16c in 41% yield according to the procedure of Example 14, Step I. ¹H NMR (CHCl₃, δ) 7.34-7.26 (m, 5H), 7.12 (t, 1H, J=7.5 Hz), 6.72-6.67 (m, 3H), 4.76-4.64 (m, 1H), 4.47 (s, 2H), 4.51-4.42 (m, 1H), 4.11-4.02 (m, 2H), 3.68 (s, 3H), 3.70-3.65 (m, 1H), 3.55-3.43 (m, 6H), 2.54-2.42 (m, 1H), 2.28-2.39 (m, 1H), 2.1-1.9 (m, 3H), 1.86-1.64 (4), 1.50-1.38 (m, 14H); ¹³C NMR (CDCl³, δ) 175.0, 172.4, 171.5, 171.1, 157.1, 138.5, 136.0, 130.1, 128.5, 127.9, 127.7, 121.0, 116.0, 114.8, 80.5, 77.6, 73.0, 66.9, 66.2, 58.2, 54.3, 52.5, 52.3, 43.4, 39.4, 39.9, 34.9, 30.0, 28.2, 26.8, 26.6, 26.0, 24.0; MS (FAB) 689 [(M+Na)⁺, 35), 667 [(M+H)⁺, 23), 633 (5), 294 (100), 204 (39), 156 (63).

[00276] The desired compound 16e was prepared from 16d according to the procedure of Example 14, Step J. The product was used without further purification.

[00277] The desired compound 16f was prepared from 16e in 20% yield according to the procedure of Example 14, Step K.¹ H NMR (CDCl₃, δ) 8.56 (d, 1H, J=7.2 Hz), 7.14 (t, 1H, J=6 Hz), 6.86 (s, 1H), 6.66 (d, 1H, J=6 Hz), 6.73 (dd, 1H, J=6.3, 15 Hz), 4.86-4.77 (m, 1H), 4.40 (dd, 1H, J=3.0, 2.7 Hz), 4.24-4.13 (m, 4 H), 3.70 (s, 3H), 3.70-3.66 (m, 2H), 3.66-3.32 (m, 3H), 2.53 (dd, 1H, J=5.7, 3.9 Hz), 2.45-2.42 (m, 1H), 1.99-1.80 (m, 6H), 1.60-1.57 (m, 4H), 1.45-1.43 (m, 11 H); ¹³C NMR (CDCl₃, δ) 175.8, 173.3, 173.0, 171.7, 160.0, 138.0, 130.1, 122.7, 116.8, 113.3, 81.0, 78.6, 66.3, 65.2, 58.8, 55.1, 52.5, 42.6, 42.4, 38.9, 34.6, 30.0, 28.2, 28.0, 27.0, 26.9, 26.8, 26.2; MS (FAB) 559 (M⁺, 33), 327 (33), 225 (100)

[00278] The desired compound 16g was prepared from 16f according to the procedure of Example 14, Step H. The product was used without further purification.

[00279] The desired compound 16h was prepared from 16g and A according to the procedure of Example 14, Step L. The product was used without further purification.

Example 17

Preparation Of Compound 17

[00281]

Example 18

Preparation of Compound of Formula 18

[00283]

[00284] To a cold (0° C.) slurry of 18a (15.0 g, 90 mmol) in dioxane (100 mL), water (100 mL), and saturated sodium bicarbonate (100 mL) was added a solution of tert-butoxycarbonyl anhydride (7.2 g, 33 mmol) in dioxane (100 mL). The reaction mixture was slowly warmed to ambient temperature over 6 hr. The reaction mixture was concentrated in vacuo. The residue was diluted with water and extracted with diethylether (2×150 mL). The ether layer was discarded. The aqueous layer was acidified slowly with solid citric acid (pH˜4) and extracted with ethyl acetate (3×150 mL). The organic layer was dried (Na₂SO₄) and concentrated in vacuo to afford 18b (14.6 g, 61% yield) as a white foam.

[00285] To a 80° C. solution of 18b (14.6 g, 54.68 mmol) in toluene (230 mL) was added DMF-di-tert-butyl acetal (53 mL, 218.72 mmol) dropwise over 2 hrs. The reaction mixture was maintained at the same temperature for 1 hr after the addition was complete. It was then cooled to ambient temperature and concentrated. The residue was purified by flash column chromatography using 96/4 to 90/10 dichloromethane/ethyl acetate to provide the required compound 18c (7.53 g, 43% yield). HRMS (FAB) Calcd for C₁₇H₂₆NO₅: 324.1811 (M+H)⁺. Found: 324.1807.

[00286] To a cold (0° C.) solution of 18c (7.5 g, 23.22 mmol) in dichloromethane (100 mL) was added triethylamine (7.12 mL, 51.08 mmol) followed by triflic anhydride (4.30 mL, 25.54 mmol) dropwise. The reaction mixture was slowly warmed to ambient temperature over 4 hrs. It was quenched with saturated bicarbonate solution and extracted into dichloromethane. The combined organic layer was washed with saturated bicarbonate and brine, dried (Na₂SO₄) and concentrated. The brown residue was purified by flash column chromatography using dichloromethane to provide 7.74 g of 18d (73% yield).

[00287] To an oven dried flask was added THF (75 mL, previously deoxygenated by bubbling argon), palladium acetate (74 mg, 0.33 mmol), R-(+)-BINAP (311 mg, 0.495 mmol), and cesium carbonate (5.38 g, 16.5 mmol) under argon atmosphere. To this mixture was added 18d (5.0 g, 11 mmol) followed by diphenylketimine (2.77 mL, 16.5 mmol). The flask was flushed with argon and heated to reflux for 12 hrs (overnight). Cooled the reaction mixture to ambient temperature and diluted with ether (500 mL). The organic layer is washed with saturated ammonium chloride solution (2×300 mL), dried (Na₂SO₄) and concentrated. Purification by flash chromatography using 100/0 to 90/10 dichloromethane/ethyl acetate provided the required product 18e (3.58 g) in 67% yield.

[00288] To a solution of 18e (3.0 g, 6.17 mmol) in methanol (62 mL) was added sodium acetate (1.218 g, 14.8 mmol), and hydroxylamine hydrochloride (0.774 g, 1 1.1 1 mmol). The reaction mixture was stirred at ambient temperature for 3 hrs. The reaction mixture was concentrated, diluted with dichloromethane and washed with 0. 1 N NaOH solution. The organic layer was dried (Na₂SO₄) and concentrated. Purification by flash column chromatography using 95/5 to 92/8 dichloromethane/ethyl acetate afforded 18f (1.31 g) in 66% yield.

[00289] To a cold (−20° C.) solution of dichloromethane (2 mL) was added chlorosulfonyl isocyanate (0.16 mL, 1.87 mmol). To this was added tert-butanol (0.18 mL, 1.87 mmol) in dichloromethane (2 mL) and slowly warmed to 0° C. over 2.5 hrs. At this time a solution of 18f (0.6 g, 1.87 mmol) in dichloromethane (6 mL) containing triethylamine (0.52 mL, 3.73 mmol) was added dropwise. The reaction mixture was warmed to ambient temperature over 12 hrs (overnight). Added saturated bicarbonate and dichloromethane and the organic layer was separated, dried (Na₂SO₄) and concentrated. Purification by flash chromatography using 95/5 to 90/10 dichloromethane/ethyl acetate provided 18g (0.59 g) in 63% yield.

[00290] The expected product 18h was synthesized as described earlier for the Example 1, Step C. The material was carried to the next step as it was.

[00291] The expected product 18i was synthesized as described earlier for the Example 1, Step D. The crude material was purified by flash chromatography using 98/2 to 90/10 dichloromethane/methanol to afford 18i in 34% yield.

[00292] The expected product 18j was synthesized as described earlier for the Example 1, Step C. The material was carried to the next step as it was.

[00293] The expected product 18k was synthesized as described earlier for the Example 1, Step J. The material after work-up was of sufficient purity to be carried forward to the next step.

Example 19

Preparation of Compound of Formula 19

[00295]

Example 20

Preparation of Compounds of Formulas 20A and 20B

[00297]

[00298] The desired product 20a was obtained by the method described for Example 1, Step F. The material was purified by flash column chromatography using 98/2 dichloromethane/methanol to provide 20a in 97% yield. HRMS (FAB) Calcd for C₃₂H₄₃N₂O₇: 567.3070 (M+H)⁺. Found: 567.3073.

[00299] The desired product 20b was obtained by the method described for Example 1, Step G. The material was purified by flash column chromatography using 98/2 to 96/4 dichloromethane/methanol to provide 20b in 81% yield. HRMS (FAB) Calcd for C₂₅H₃₇N₂O₇: 477.2601 (M+H)⁺. Found: 477.2606.

[00300] The desired product 20c was obtained by the method described for Example 1, Step H. Purification by column chromatography using 99/1 dichloromethane/methanol afforded 20c along with triphenylphosphine oxide. This mixture was taken to the next step. HRMS (FAB) Calcd for C₂₅H₃₅N₂O₆: 459.2495 (M+H)⁺. Found: 459.2490.

[00301] The desired product 20d was obtained by the method described for Example 1, Step I. Yield of 20d (for 2 steps) 23%. ¹H NMR (DMSO-d₆). 0.84 (m, 2H), 1.10 (m, 3H), 1.56-1.67 (m, 6H), 1.75-1.81 (m, 1H), 2.32-2.49 (m, 3H), 2.55-2.59 (m,1H), 2.94 (dt,1H), 3.50 (dd,1H), 3.56-3.65 (m, 2H), 3.99 (dd,1H), 4.06-4.23 (m, 4H), 4.37 (t,1H), 6.64-6.74 (m, 3H), 7.08 (app. t, 1H), 7.95 (d, 1H), 12.30 (br. s, 1H); ¹³C NMR δ (DMSO-d₆) 25.25, 25.97, 28.30, 28.55, 30.61, 33.77, 36.04, 39.41, 52.52, 54.02, 57.22, 66.38, 68.03, 77.49, 114.75, 115.37, 121.14, 128.86, 142.66, 158.92, 169.87, 170.83, 172.99; HRMS (FAB) Calcd for C₂₄H₃₃N₂O₆: 445.2339 (M+H)⁺. Found: 445.2343.

[00302] The expected product 20e was synthesized as described earlier for the Example 1, Step J. The material after work-up was of sufficient purity to be carried forward to the next step.

Example 21

[00304] Preparation of Compound of Formula 21

[00305] The expected product 21a was synthesized as described earlier for the Example 2, Step A. The material after work-up was of sufficient purity to be carried forward to the next step.

Example 22

Preparation of Compound of Formula 22

[00307]

Example 23

Preparation of Compound of Formula 23

[00309]

[00310] The desired compound 23a was prepared in 58% yield from 1I and D according to the method of Example 1, Step J.

Example 24

Preparation of Compound of Formula 24

[00312]

Example 25

Preparation of Compound of Formula 25

[00314]

Example 26

Preparation of Compounds of Formulas 26A and 26B

[00316]

[00317] To a cold (0° C.) solution of 26a (4.0 g, 20 mmol) in THF/MeCN (35/5 mL) was added 4-nitrophenyl chloroformate (4.86 g, 24 mmol) and then pyridine (1.9 mL, 24 mmol). The reaction mixture was warmed to ambient temperature over 4.5 hrs. Reaction was monitored till consumption of 26a (needed to add some more of other two reagents). The reaction was quenched by adding water, the organics were separated, washed with brine, dried (Na₂SO₄), and evaporated in vacuo to afford the product 26b. This material was sufficiently pure for further studies.

[00318] To a solution of 1f (2.3 g, 5.1 mmol) in dichloromethane/DMF (25/5 mL) at 0° C. was added 26b (2.24 g, 6.1 mmol) followed by triethylamine (0.86 mL, 6.1 mmol). Few crystals of imidazole were added and the reaction mixture stored at −8° C. for 16 hrs. The reaction mixture was diluted with dichloromethane, washed with saturated sodium bicarbonate, 10% aqueous citric acid solution, dried (Na₂SO₄), and evaporated in vacuo. The crude material was purified by flash column chromatography using 100/0 to 70/30 dichloromethane/ethyl acetate to provide 26c (1.2 g, 38% yield). HRMS (FAB) Calcd for C₃₈H₄₈N₃O₇: 658.3492 (M+H)⁺. Found: 658.3483.

[00319] The desired product 26d was obtained by the method described for Example 1, Step G. The crude material was carried to the next step as it was. HRMS (FAB) Calcd for C₂₄H₃₆N₃O₇: 478.2553 (M+H)⁺. Found: 478.2547.

[00320] The desired product 26e was obtained by the method described for Example 1, Step H. Purification by column chromatography using 99/1 dichloromethane/methanol afforded 26e along with triphenylphosphine oxide. This mixture was taken to the next step.

[00321] The desired product 26f was obtained by the method described for Example 1, Step I. HRMS (FAB) Calcd for C₂₃H₃₂N₃O₆: 446.2291 (M+H)⁺. Found: 446.2290.

[00323] The desired product 21 was obtained by the oxidation protocol described previously for Example 2, Step B. Purification by flash column chromatography using 100/0 to 98/2 dichloromethane/methanol afforded 21 in 38% yield. HRMS (FAB) Calcd for C₄₄H₆₀N₅O₁₀: 818.4340 (M+H)⁺. Found: 818.4329. chromatography using 98/2 dichloromethane/methanol to provide 26g in 31% yield (3 steps). HRMS (FAB) Calcd for C₄₁H₅₈N₇O₉: 792.4296 (M+H)⁺. Found: 792.4284.

Example 27

Preparation of Compound 27

[00325]

[00326] To the mixture of Boc-3,4-didehydroproline-OMe (27a, 5.30 g, 23.4 mmol), N-methylmorpholine-N-oxide (4.75 g, 35.1 mmol) in acetone (10 mL) and water (15 mL) at rt. was added an Osmium tetroxide solution in tert-butanol (2.5% w/w, 3.5 mL, 0.344 mmol). To this cloudy solution was added THF until the mixture became almost homogeneous. After stirred at rt. overnight, saturated aqueous sodium thiosulfate solution (30 mL) was added and 10 min later, followed by addition of EtOAc (300 mL) and brine (80 mL). After layers were separated, aqueous solution was extracted with EtOAc (2×100 mL). Organic solutions were combined, dried (MgSO₄), filtered and concentrated in vacuo to give a dark liquid. Flash chromatography (4 to 8% MeOH/CH₂Cl₂) afforded 27b (4.73 g, 18.1 mmol, 77%) as an oil.

[00327] To a suspension of the diol 27b (1.6 g, 6.12 mmol) magnesium sulfate (4.0 g, 33.2 mmol) and 3-benzyloxypropionaldehyde (2.32 g, 13.0 mmol) in anhydrous CH₂Cl₂ (60 mL) at 0° C. was added p-toluenesulfonic acid (150 mg, 1.01 mmol). The resulting mixture was stirred vigorously and allowed to warm to rt. along with the ice-bath overnight (18 h). Saturated sodium bicarbonate solution (60 mL), water (30 mL) and CH2Cl2 (100 mL) were added and the layers were separated. The aqueous solution was extracted with CH₂Cl₂ (2×100 mL) and combined organic solution was dried (MgSO4), filtered and concentrated in vacuo to give a colorless oil. Flash chromatography (5 to 15 % EtOAc/CH₂Cl₂) afforded 27c (2.35 g, 5.57 mmol, 91%) as an oil.

[00328] The desired compound 27d was prepared from 27c according to the method of Example 1, Step C. It was used in the next reaction without further purification.

[00329] The desired compound was prepared from 27d according to the method of Example 1, Step D. Flash chromatography (8 to 20% EtOAc/CH₂C₁₂) afforded 27e.

[00330] The desired compound 27f was prepared from 27e according to the method of Example 1, Step E. It was used in the next reaction without further purification.

[00331] The desired compound was prepared from 27f according to the method of Example 1, Step F. Flash chromatography (8 to 20% EtOAc/CH₂Cl₂) afforded 27g (36%, 4 steps). HRMS m/z 595.3014 [calcd for C₃₃H₄₂N₂O₈, 595.3019].

[00332] The desired compound was prepared quantitatively from 27g according to the method of Example 1, Step G. Flash chromatography (3 to 5% MeOH/CH₂Cl₂) afforded 27h as a white solid. HRMS m/z 595.2553 [calcd for C₂₆H₃₆N₂O₈, 595.2550].

[00333] The desired compound was prepared from 27h according to the method of Example 1, Step H. Flash chromatography (3 to 5% MeOH/CH₂Cl₂) afforded 27i as a mixture with triphenylphosphine oxide which was hydrolyzed.

[00334] The desired compound was prepared (72%, 2 steps) from 27i according to the method of Example 1, Step I.

[00335] The desired compound was prepared from 27j according to the method of Example 1, Step J. Flash chromatography (3 to 6% MeOH/CH₂Cl₂) afforded 27k (69%) as a mixture of diastereomers.

Example 28

Preparation of Compound 28

[00337]

[00338] The desired compound was prepared from 27j according to the method of Example 2, Step A. Flash chromatography (3 to 6% MeOH/CH₂Cl₂) afforded 28a (50%) as a mixture of inseparable diastereomers.

Example 29

Preparation of Compound 29

[00340]

[00341] The desired compound 29A was prepared from 28A according to the method of Example 3, Step A.

Example 30

Preparation of Compound 30

[00343]

[00344] To a solution of the Cbz—HYP—OMe (30a) (3.0 g, 10.7 mmol) and 4-benzyloxy-2-methyl-1-butene 30b (5.30 g, 30.0 mmol) in anhydrous CH₂Cl₂ (80 mL) at 0° C. was added trifluoroboron diethyl etherate (0.25 mL, 1.97 mmol). The resulting mixture was stirred at rt. overnight (18 h). Saturated sodium bicarbonate solution (30 mL), brine (50 mL) and EtOAc (300 mL) were added and the layers were separated. The aqueous solution was extracted with EtOAc (2×100 mL) and the combined organic solution was dried (MgSO₄), filtered and concentrated in vacuo to give a yellow oil. Flash chromatography (5 to 20% EtOAc/CH₂Cl₂) afforded 30c (2.00 g, 4.39 mmol, 41%) as an oil.

[00345] The desired compound 30d was prepared from 30c quantitatively according to the procedure of Example 1, Step G.

[00346] The desired compound 30e was prepared from 30d and Boc-cyclohexylglycine-OH according to the procedure of Example 1, Step D. Flash chromatography (3 to 5% MeOH/CH₂Cl₂) afforded 30e (61%).

[00347] The solution of 30e and 2 N HCl in dioxane and EtOAc (1:1) was stirred at rt. for 3 h, after which it was concentrated in vacuo. The product was used in the next reaction without further purification.

[00348] The desired compound 30g was prepared from 30f according to the procedure of Example 1, Step F. Flash chromatography (2 to 5% MeOH/CH₂Cl₂) afforded 30g (48%, 2 steps).

[00349] The mixture of 30g (700 mg, 1.28 mmol) and potassium carbonate (530 mg, 3.84 mmol) in anhydrous methanol (80 mL) was vigorously stirred at rt. The reaction progress was monitored by TLC. After 3 h, it was concentrated in vacuo before EtOAc (200 mL) and water (100 mL) were added and layers were separated. The aqueous solution was extracted with EtOAc (2×100 mL). The organic solutions were combined, dried (MgSO₄), filtered and concentrated in vacuo. The product was used in the next reaction without further purification.

[00350] The desired compound 30i was prepared from 30h according to the procedure of Example 1, Step H.

[00351] The desired compound 30j was prepared from 30i according to the procedure of Example 1, Step I (23%, 3 steps).

[00352] The desired compound was prepared from 30j according to the procedure of Example 1, Step J . Flash chromatography (3 to 6% MeOH/CH₂Cl₂) afforded 30k (58%).

Example 31

Preparation of Compound 31

[00354]

[00355] The desired compound was prepared from 30j and B according to the procedure of Example 1, Step J. Flash chromatography (2 to 5% MeOH/CH₂Cl₂) afforded 31a (73%).

Example 32

Preparation of Compound 32

[00357]

Example 33

Preparation of Compound 33

[00359]

[00360] The suspension of methyl 3,5-dihydroxyphenylacetate (33a) (5.0 g, 27.4 mmol) methyl iodide (4.6 g, 32.9 mmol), potassium carbonate (5.69 g, 41.2 mmol) in DMF (30 mL) was heated to 55° C. and stirred overnight. After cooling to rt., saturated aqueous sodium bicarbonate solution (100 mL) and EtOAc (200 mL) were added and layers were separated. The aqueous solution was extracted with EtOAc (2×100 mL). The combined organic solution was dried (MgSO₄), filtered and concentrated in vacuo. Flash chromatography (2 to 5% MeOH/CH₂Cl₂) afforded 33b (1.11 g, 29%) as a white liquid.

[00361] An aqueous lithium hydroxide solution (0.342 g in 10 mL H₂O) was added to a solution of the methyl ester 33b in THF (10 mL) and methanol (10 mL) at rt. The progress of the reaction was monitored by TLC. After 4 hr., the volatiles were removed in vacuo, EtOAc (150 mL) and water (30 mL) were added and the aqueous solution was acidified to pH=1 and saturated with solid sodium chloride. After separation of the layers, the aqueous layer was extracted with EtOAc (2×150 mL). Organic solutions were combined, dried with sodium sulfate, filtered and concentrated in vacuo to afford 33c (1.6 g).

[00362] The desired compound 33d was prepared from 33c and 1f according to the procedure of Example 1, Step F. Flash chromatography (2 to 5% MeOH/CH₂Cl₂) afforded 33d in 90% yield.

[00363] The desired compound 33e was prepared from 33d according to the procedure of Example 1, Step G. Flash chromatography (2 to 5% MeOH/CH₂Cl₂) afforded 33e in 56% yield.

[00364] The desired compound 33f was prepared from 33e according to the procedure of Example 1, Step H. Flash chromatography (2 to 5% MeOH/CH₂Cl₂) afforded 33e as a mixture with triphenylphosphine oxide which was used in the next reaction without further purification.

[00365] The desired compound 33g was prepared from 33f in 45% yield (2 steps) according to the procedure of Example 1, Step I.

[00366] The desired compound 33h is prepared from 33g and A according to the procedure of Example 1, Step J.

Example 34

Preparation of Compound 34

[00368]

[00369] The desired compound 34a is prepared according to the method of Example 1, Step J, except substituting amine B for amine A. The product is obtained as a mixture of inseparable diastereomers.

Example 35

Preparation of Compound 35

[00371]

Example 36

Preparation of Compound 36

[00373]

[00374] A solution of tert-butyl phosphonoacetate (15.1 g, 50.0 mmol) in dry THF was cooled to 0° C. and treated with NaH (60%, 2.5 g, 62.5 mmol, 1.25 equiv.) and stirred for 20 min. The reaction mixture was treated with 2-pentanone (4.3 g, 50 mmol) and stirred at rt. for 24 h. The reaction mixture was diluted with aq. NaHCO₃ and extracted into ether (3×100 mL). The combined ether layer were extracted with brine, dried (MgSO₄), concentrated in vacuo and distilled to yield 8.2 g (88%) of 36b (stereochemical ratio 2:1).

[00375] A solution of 36b (5.0 g, 27.1 mmol) was dissolved in dichloroethane and treated with 4,4′-thiobis-(2-tert-butyl-5-methylphenol) (100 mg) and MCPBA (60-80%, 7.76 g, 27.1 mmol) and heated at reflux for 4 h. The reaction mixture was concentrated in vacuo and the residue was diluted with ether (200 mL) The ether layer was washed twice with satd aq Na₂S₂O₃, aq. NaOH and brine (100 ml). The reaction mixture was concentrated in vacuo to yield 4.2 g (77%) of 36c which was used as it is in the next step.

[00376] A solution of epoxide (4.0 g, 18.4 mmol) in dry ethanol (100 mL) was treated with NaN₃ (12 g, 184 mmol) and NH₄Cl (9.6 g. 184 mmol) and heated at reflux for 36 h. The reaction mixture was diluted with water and the reaction mixture was extracted with ether (300 mL). The combined organic layers were dried with Na₂SO₄ and concentrated. The residue was purified by SiO₂ chromatography (EtOAc/Hex 1:19) to yield 1.1 g (28%) of 36d and 731 mg (18%) of 36d′ as a colorless liquid.

[00378] A solution of Cbz—Cl (1.48 g, 8.7 mmol, 1.23 mL) was added dropwise to a mixture of amine and Et3N (878 mg, 1.25 mL) at −78° C. in CH₂Cl₂ (30 mL). The reaction mixture was warmed to rt. and concentrated in vacuo. The residue was chromatographed on SiO₂ (EtOAc/Hex 8:2) to 36e (450 mg, 15%) as a colorless solid.

[00380] The acid obtained by hydrolysis of 36e was dissolved in CH₂Cl₂ (10 mL) at −20° C. and treated with H-Glycyl-Phenylglycycl-N(CH₃)₂ (281 mg, 0.93 mmol), HOOBt (208 mg, 1.27 mmol, 1.25 equiv.) EDCl (244 mg, 1.27 mmol) and NMM (343 mg, 3.4 mmol, 490 μL). The reaction mixture was stored in the freezer for 24 h and diluted with aq. HCl (1 M, 50 mL). The reaction mixture was extracted with CH₂Cl₂ (3×50 mL). The combined organic layers were extracted with aq. HCl (1M, 100 mL), aq. NaHCO₃ (1M, 100 mL), brine (100 ml), dried (MgSO₄), filtered and concentrated in vacuo and chromatographed on SiO₂ (acetone/Hexanes 1:3) to yield 36f (330 mg, 75%) as a colorless solid.

[00381] A solution of 36f is dissolved in CH₃OH (20 mL) and treated with Pd/C reaction mixture will be hydrogenated at 40 psi for 12 filtered through a plug of Celite and the filtrate is directly used in the next step.

[00382] The expected product 36h is synthesized as described earlier for the Example 1, Step J. The coupled material will be used directly for the next step to synthesize 36A and 36 B.

Example 37

Preparation of Compound 37

[00384]

[00385] To a cold (0° C.) solution of F (2.3 g g, 9.43 mmol) in dichloromethane (20 mL) was added triethylamine (3.97 mL, 28.28 mmol), DMAP (few crystals) and 3-bromobenzenesulfonyl chloride (3.61 g, 14.14 mmol). The reaction mixture was left standing in the refrigerator (0°-5° C.) overnight. The reaction mixture was washed with saturated NaHCO₃, and 10% citric acid solution. The organic layer was dried (Na₂SO₄) and concentrated. The residue was purified by flash chromatography using 95/5 to 90/10 dichloromethane/EtOAc to afford 2.7 g (62% yield) of 37a. HRMS (FAB) Calcd for C₁₇H₂₄N₂O₆SBr: 465.0518 (M+H)⁺. Found: 465.0497.

[00386] The desired product 37b was obtained by the method described for Example 1, Step C. The crude material was used in the next step without purification.

[00387] The desired product 37c was obtained by the method described for Example 1, Step D in 97% yield. The material after workup was pure enough to be carried forward.

[00388] The desired product 37d was obtained by the method described for Example 1, Step E. The crude material was used in the next step without purification.

[00389] The desired product 37e was obtained by the method described for Example 1, Step F using pentenoic acid as the coupling partner. Purification of the residue by column chromatography using 90/10 to 80/20 dichloromethane/EtOAc provided 35% yield of 37e. HRMS (FAB) Calcd for C₂₅H₃₅N₃O₆SBr: 586.1409 (M+H)⁺. Found: 586.1423.

[00390] To a stirred solution of 37e (660 mg, 1.13 mmol) in DMF (10 mL) under nitrogen atmosphere was added triethylamine (3.61 mL, 32.77 mmol), potassium carbonate (780 mg, 5.65 mmol), tetrabutylammonium bromide (730 mg, 2.26 mmol), and palladium (II) acetate (33 mg, 0.15 mmol). the mixture was heated at 100° C. for 2 hrs. The reaction mixture was cooled to room temperature, diluted with EtOAc and washed with 5% phosphoric acid solution. The organic layer was separated, dried (Na₂SO₄) and concentrated to provide 280 mg (49% yield) of 37f as a mixture of diastereomers. This material was pure enough to be carried to the next step. HRMS (FAB) Calcd for C₂₅H₃₄N₃O₆S: 504.2168 (M+H)⁺. Found: 504.2155.

[00391] The desired product 37g was obtained by the hydrogenation procedure described for Example 1, Step G in 73% yield. The material was sufficiently pure for further studies. HRMS (FAB) Calcd for C₂₅H₃₆N₃O₆S: 506.2325 (M+H)⁺. Found: 506.2314.

[00392] The desired product 37h was obtained by the procedure described for Example 1, Step H in 84% yield. HRMS (FAB) Calcd for C₂₄H₃₄N₃O₆S: 492.2168 (M+H)⁺. Found: 492.2175.

[00393] The expected product 37i was obtained by the procedure described for Example 2, Step A in 90% yield. The crude material was sufficiently pure for further studies. HRMS (FAB) Calcd for C₄₄H₆₃N₆O₁₀S: 867.4326 (M+H)⁺. Found: 867.4342.

Example 38

Preparation of Compound 38

[00395]

[00396] The desired material 38A was synthesized as described for Example 3, Step A in 91% yield.

Example 39

Preparation of Compound 39

[00398]

[00399] The desired product 39a was obtained by the method described for Example 37, Step A using pipsyl chloride instead of 3-bromobenzenesulfonyl chloride. Purification of the residue by column chromatography using 95/5 to 90/10 dichloromethane/EtOAc provided 75% yield of 39a. HRMS (FAB) Calcd for C₁₇H₂₄N₂O₆Sl: 511.0400 (M+H)⁺. Found: 511.0386.

[00400] The desired product 39b was obtained by the method described for Example 1, Step C. The crude material was used in the next step without purification.

[00401] The desired product 39c was obtained by the method described for Example 1, Step D. Purification of the residue by column chromatography using 90/10 to 80/20 dichloromethane/EtOAc provided 68% yield of 39c. HRMS (FAB) Calcd for C₂₅H₃₇N₃O₇Sl: 650.1397 (M+H)⁺. Found: 650.1398.

[00402] The desired product 39d was obtained by the method described for Example 1, Step E. The crude material was used in the next step without purification.

[00403] The desired product 39e was obtained by the method described for Example 1, Step F using pentenoic acid as the coupling partner. Purification of the residue by column chromatography using 98/2 dichloromethane/MeOH provided 76% yield of 39e.

[00404] The desired product 39f was obtained by the method described for Example 37, Step F. Purification of the residue by column chromatography using 98/2 dichloromethane/MeOH provided 28% yield of 39f as a mixture of diastereomers. HRMS (FAB) Calcd for C₂₅H₃₄N₃O₆S: 504.2168 (M+H)⁺. Found: 504.2160.

[00405] The desired product 39g was obtained by the hydrogenation procedure described for example 1, Step G in 84% yield. The material was sufficiently pure for further studies. HRMS (FAB) Calcd for C₂₅H₃₆N₃O₆S: 506.2325 (M+H)⁺. Found: 506.2314.

[00406] the desired product 39h was obtained by the procedure described for Example 1, Step H in quantitative yield. HRMS (FAB) Calcd for C₂₄H₃₄N₃O₆S: 492.2168 (M+H)⁺. Found: 492.2175.

[00407] The expected product 39i was obtained by the method described for Example 2, Step A in 36% yield. The crude material was sufficiently pure for further studies. HRMS (FAB) Calcd for C₄₄H₆₃N₆O₁₀S: 867.4326 (M+H)⁺. Found: 867.4342.

Example 40

Preparation of Compound 40

[00409]

Example 41

Preparation of Compound 41

[00411]

[00412] To a 25 mL addition funnel was added benzene (5 mL), DMF (0.32 mL, 4.1 mmol), and thionyl chloride (0.33 mL, 4.5 mmol). After 5 minutes two layers appeared. The lower layer was separated and added slowly to a cooled (0-5° C.) solution of acrylic acid (0.19 mL, 2.8 mmol) in dichloromethane. The mixture was maintained at that temperature for 10 minutes. Then triethylamine (0.77 mL, 5.5 mmol) was added followed by 39d (1.13 g, 2.1 mmol). The reaction mixture was warmed to ambient temperature over 5 hrs and quenched with saturated NaHCO₃. The organic layer was separated, washed with 5% H₃PO₄ solution and brine. The dichloromethane layer was dried (Na₂SO₄), and concentrated. The crude material was purified by flash column chromatography using 98/2 dichloromethane/MeOH to provide 870 mg of 41a (67% yield). HRMS (FAB) Calcd for C₂₃H₃₁N₃O₆Sl: 604.0978 (M+H)⁺. Found: 604.0964.

[00413] The desired product 41b was obtained by the method described for Example 37, Step F. The residue was purified using 97/3 dichloromethane/MeOH to afford 41b in 26% yield. HRMS (FAB) Calcd for C₂₃H₃₀N₃O₆S: 476.1855 (M+H)⁺. Found: 476.1858.

[00414] The desired product 41c was obtained by the hydrogenation method described for Example 1, Step D in 75% yield. The material was sufficiently pure for further studies.

[00415] The desired product 41d was obtained by the method described for Example 1, Step E. The crude material was used in the next step without purification.

[00416] The expected product 41e was obtained by the method described for Example 2, Step A in 63% yield. The crude material was sufficiently pure for further studies.

Example 42

Preparation of Compound 42

[00418]

Example 43

Preparation of Compound 43

[00420]

[00421] To a cold (0° C.) solution of F (5.4 g, 22.1 mmol) in dichloromethane (50 mL) was added triethylamine (6.8 mL, 48.6 mmol), DMAP (few crystals) and benzenesulfonyl chloride (3.29 g, 24.1 mmol). The reaction mixture was left standing in the refrigerator (0-5° C.) overnight. The reaction mixture was washed with saturated NaHCO₃, and 10% citric acid solution. The organic layer was dried (Na₂SO₄) and concentrated. The residue was purified by flash chromatography using 95/5 dichloromethane/EtOAc to afford 5.0 g (59% yield) of 43a.

[00422] The desired product 43b was obtained by the method described for Example 1, Step C. The crude material was used in the next step without purification.

[00423] The desired product 43c was obtained by the method described for Example 1, Step D. Purification of the residue by column chromatography using 99/1 dichloromethane/MeOH provided 60% yield of 43c.

[00424] Argon gas was bubbled into a cold (20° C.) solution of 43c (1.72 g, 3.28 mmol) in dichloromethane (40 mL) for 20-30 min. ADDP (2.5 g, 9.84 mmol) was added followed by triphenylphosphine (2.6 g, 9.84 mmol) and 3-benzyloxypropanol (0.57 mL, 3.61 mmol). The reaction was warmed to ambient temperature and left standing for 2 days. Concentrated the reaction mixture and added Et₂O (50 mL). The precipitated solid material was filtered off. Repeated this operation twice to remove most of the side products. The filtrate was concentrated and purified by flash chromatography using 90/10 to 85/15 dichloromethane/EtOAc to provide 330 mg of 43d. The recovered starting materials, along with some triphenylphosphine oxide, was resubjected to the above described conditions to provide additional 420 mg of 43d. Combined yield=34%. HRMS (FAB) Calcd for C₃₅H₅₀N₃O₈S: 672.3319 (M+H)⁺. Found: 672.3330.

[00425] The desired product 43e was obtained by the method described for Example 1, Step E. The crude product was carried to the next step.

[00426] The desired compound 43f was prepared by the protocol described for Example 1, step F. Purification by flash chromatography was carried out with 60/40 to 50/50 dichloromethane/EtOAc tp provide 43f in quantitative yield. HRMS (FAB) Calcd for C₃₈H₄₈N₃O₈S: 706.3162 (M+H)⁺. Found 706.3157.

[00427] The desired product 43g was obtained by the procedure described for Example 1, Step G. The material was sufficiently pure for further studies.

[00428] The desired product 43h was obtained by the procedure described for Example 1, Step H. After completion of reaction, the solvent was evaporated. Et₂O (50 mL) was added and the solids were filtered off. The filtrate was concentrated and Et2O/EtOAc (50 mL/50 mL) was added. The precipitated solids were filtered off and the filtrate was concentrated. The residue was purified by flash chromatography using 85/15 to 80/20 dichloromethane/EtOAc to afford pure 43h in 20% yield. HRMS (FAB) Calcd for C₃₁ H₄₀N₃O₇S: 598.2587 (M+H)⁺. Found: 598.2581.

[00429] The expected product 43i was obtained by the procedure described for Example 1, Step I in 92% yield.

[00430] The desired material 43j was synthesized as described for Example 1, Step J. The crude product was of sufficient purity to be carried forward.

Example 44

Preparation of Compound 44

[00432]

[00433] The desired product 44a was obtained by the method described for Example 1, Step D using N-boc-tert-butylglycine as the coupling partner. The material after work-up was sufficiently pure to be carried to the next step.

[00434] The desired product 44b was obtained by the method described for Example 1, Step E. The crude material was carried forward without purification.

[00435] The desired product was obtained by the method described for Example 1, Step F, using 3-hydroxybenzoic acid. The material was purified by flash column chromatography using 85/15 to 65/35 dichloromethane/ethyl acetate to provide 44c in 81% yield.

[00436] The desired product 44d was obtained by the method described for Example 1, Step G. The residue after workup was sufficiently pure for further manipulation.

[00437] The desired product 44e was obtained by the method described for Example 1, Step H. The crude residue after concentration was taken in hexanes/EtOAc (1/1) and the solid material was filtered off. This operation was performed again to remove some of the side products. Purification by column chromatography using 80/20 dichloromethane/EtOAc afforded 44e along with triphenylphosphine oxide. This mixture was taken to the next step

[00438] The desired product 44f was obtained by the method described for Example 1, Step I. Yield of 44f (for 2 steps)=11%.

[00439] The expected product 44g was synthesized as described earlier for the Example 1, Step J. The material after work-up was carried forward to the next step.

Example 45

Preparation of Compound 45

[00441]

[00442] Alkylation of 45a with 45b was carried out using the procedure described for Example 1 Step A. The crude product was purified using 85/0/15 to 85/5/10 hexanes/EtOAc/dichloromethane to provide 45c in 37% yield. HRMS (FAB) Calcd for C₁₈H₂₉O₄: 309.2066 (M+H)⁺. Found: 309.2060.

[00443] To a solution of 45c (4.8 g, 15.5 mmol) in MeOH (30 mL) was added pyridinium p-toluenesulfonate (780 mg, 3.1 mmol) and refluxed for 3 hrs when all the starting material was consumed. The reaction mixture was concentrated. The residue was dissolved in EtOAc, washed with saturated NaHCO₃, dried (Na2SO4) and the organic layer was concentrated to afford 3.2 g of 45d (92% yield). This material was sufficiently pure for further studies. HRMS (FAB) Calcd for C₁₃H₂₁O₃: 225.1491 (M+H)⁺. Found: 225.1486.

[00444] The desired product 45e was obtained by the procedure described for Example 10 Step A using 45d as the starting material. The crude product was purified using 90/10 hexanes/EtOAc to provide 45e in 70% yield. HRMS (FAB) Calcd for C₁₉H₂₄O₅SBr: 443.0528 (M+H)⁺. Found: 443.0552.

[00445] The desired product 45f was obtained by the procedure described for Example 1 Step A using 45e as the starting material. The crude product was carried further without purification.

[00446] The desired product 45g was obtained by the procedure described for Example 18 Step B using 45f as the starting material. The residue was purified by flash chromatography using 90/10 dichloromethane/EtOAc to provide 45g in 61% yield (for 2 steps). HRMS (FAB) Calcd for C₂₇H₄₄NO₇: 494.3118 (M+H)⁺. Found: 494.3107.

[00447] The desired compound 45h was prepared by the protocol described for Example 1 Step C. The material was carried forward.

[00448] The desired product 45i was obtained by the procedure described for Example 1 Step D. The residue was purified by flash chromatography using 90/10 to 85/15 dichloromethane/EtOAc to provide 45i in 41% yield.

[00449] The desired product 45j is obtained by the hydrogenation protocol described previously for Example 1 Step G.

[00450] The desired product 45k is obtained by the procedure described previously for Example 1 Step C.

[00451] To a cold (0° C.) solution of 45k in dichloromethane is added triethylamine followed by carbonyldiimidazole. Slow warming to ambient temperature overnight is expected to provide the required product 45l. This product can be purified using conventional flash column chromatography to afford pure 45l.

[00452] The desired product 45m is obtained by the producer described previously for Example 3 Step A.

[00453] The expected product 45n is synthesized as described earlier for Example 1 Step J.

Example 46

Preparation of Compound 46

[00455]

[00456] To a solution of 46a (10.0 g, 42.9 mmol) in dichloromethane (100 mL) was added BOP (22.75 g, 51.5 mmol) and stirred at room temperature for 10 minutes. N,O-dimethylhydroxylamine hydrochloride (4.18 g, 42.9 mmol) was then added followed by triethylamine (18.1 mL, 128.7 mmol). The reaction mixture was stirred at ambient temperature for 3 hours and then washed with 3N HCl, saturated NaHCO₃, and brine. The organic layer was dried (Na₂SO₄) and concentrated. The crude material, 46b, was used in the following step without purification.

[00457] To a cold (0° C.) solution of 46b in THF (60 mL) was added a solution of LAH (1M in THF, 50 mL, 50 mmol) under nitrogen atmosphere. Reaction was kept at that temperature for 30 min. The reaction mixture was quenched (slow addition) with aqueous 10% potassium hydrogen phosphate solution (30 mL). The mixture was extracted with EtOAc twice. The combined organic layer was washed with 3N HCl, saturated NaHCO₃, and brine. Dried the EtOAc layer over sodium sulfate and concentrated. The residue was purified by flash column chromatography using 10/90 to 30/70 EtOAc/dichloromethane to provide 4.2 g of 46c (45% yield—for 2 steps). HRMS (FAB) Calcd for C₁₀H₂₀NO₃S: 234.1164 (M+H)⁺. Found: 234.1168.

[00458] To a cold (−15° C.) solution of 46c (4.2 g, 19.4 mmol) in dichloromethane was added acetic acid (2.14 mL, 38.8 mmol) followed by methyl isocyanoacetate (1.76 mL, 19.4 mmol). The reaction mixture was warmed to room temperature and let stand for 16 hrs. Diluted the reaction with EtOAc and washed with saturated NaHCO₃, brine and water. The organic layer was dried (Na2SO₄) and concentrated. Purification by flash column chromatography using 30/70 EtOAc/dichloromethane afforded pure 46d (6.5 g) as a white solid in 92% yield. HRMS (FAB) Calcd for C₁₆H₂₉N₂O₇S: 393.1695 (M+H)⁺. Found: 393.1692.

[00459] To a solution of 46d (6.5 g, 16.6 mmol) in MeOH (30 mL) was added a solution of lithium hydroxide (1.19 g, 50 mmol) in water (30 mL). After 45 minutes the reaction mixture was concentrated. Aqueous citric acid solution was added till acidic pH (3) and the product was extracted into EtOAc. Concentration of the organic layer resulted in the acid 46e (5.6 g, 90% yield). HRMS (FAB) Calcd for C₁₃H₂₅N₂O₆S: 337.1433 (M+H)⁺. Found: 337.1430.

[00460] The desired product 46f was obtained by the method described for Intermediate A, Step 3. Purification of the residue by column chromatography using 20/0/80 to 50/5/45 of EtOAc/NH₃ in MeOH/dichloromethane afforded 3.0 g of 46f (51%). HRMS (FAB) Calcd for C₂₃H₃₇N₄O₆S: 497.2434 (M+H)⁺. Found: 497.2439.

[00461] The desired compound 46g was prepared by the protocol described for Intermediate A, Step 4. The material was carried forward for further studies.

[00462] The desired product 46h was obtained by the procedure described for Example 1, Step J. The crude material was sufficiently pure for further manipulations. HRMS (FAB) Calcd for C₄₀H₅₇N₆O₉S: 797.3908 (M+H)⁺. Found: 797.3896.

Example 47

Preparation of Compound 47

[00464]

Example 48

Preparation of Compound 48

[00466]

[00467] The desired product 48b was obtained by the procedure described for Example 1 Step J. The crude material was sufficiently pure for further manipulations. (Note: The precursor 48a was obtained from commercially available Nboc-S-methylcysteine by similar procedures described for 46g)

Example 49

Preparation of Compound 49

[00469]

Example 50

Preparation of Compound 50

[00471]

[00472] The desired product 50a is obtained by the method described for Example 1 Step F, using imidazole-4-acetic acid as the coupling partner.

[00473] The desired product 50b is obtained by the method described for Example 1 Step G.

[00474] The desired product 50c is obtained by the method described for Example 10 Step A using p-toluenesulfonyl chloride as the starting material.

[00475] The desired product 50d is obtained by the treating 50c with HOBt in THF at ambient temperature over several hrs.

[00476] The expected product 50e is synthesized by heating of 50d with sodium carbonate, and sodium iodide in acetone at 50° C. over several hrs. The product can be purified by conventional flash chromatography.

[00477] The desired products 50f is obtained by the hydrolysis protocol described previously for Example 1 Step I.

[00478] The expected product 50g is synthesized as described earlier for Example 1 Step J.

Example 51

Preparation of Compound 51

[00480]

[00481] The expected product 51a is synthesized as described earlier for Example 2 Step A.

Example 52

Preparation of Compound 52

[00483]

Example 53

Preparation of Compounds of Formulas 53A and 53B

[00485]

[00486] The desired product 53a was obtained by the method described in Example 1, Step F. The material was purified by flash column chromatography using 80/20 to 60/40 dichloromethane/ethyl acetate to provide 53a.

[00487] The desired product 53b was obtained by the method described in Example 1, Step G. The crude material was carried to the next step as is.

[00488] The desired product 53c was obtained by the method described in Example 1, Step H. Purification by column chromatography using 99/1 dichloromethane/methanol afforded 53c along with triphenylphosphine oxide. This mixture was taken to the next step.

[00489] The desired product was obtained by the method described in Example 1, Step 1.

[00490] The expected product 53e was synthesized as described earlier for the Example 1, Step J. The material after work-up was of sufficient purity to be carried forward to the next step.

Example 54

Preparation of Compound 54

[00492]

[00493] Commercially available 54a was converted to the desired product 54b using the procedure described for Intermediate A Step 3 in quantitative yield.

[00494] To a cold (0° C.) solution of 54b (8 g, 26.8 mmol) in DMF (100 mL) was added sodium hydride (60% dispersion in oil, 1.3 g, 32.16 mmol). After 10 minutes iodomethane (2.8 mL, 42.8 mmol) was added and the reaction was warmed to ambient temperature over 2 hrs. The reaction mixture was quenched with aqueous NH₄Cl solution and extracted with EtOAc. The organic layer was separated, dried (Na₂SO₄) and concentrated to afford 54c which was sufficiently pure for further studies.

[00495] The desired product 54d was obtained by the procedure described for Example 1 Step C. The crude product was used without further purification.

[00496] The desired product 54f was obtained by the procedure described for Example 26 Step A using 54e as the starting material. The crude product was purified by flash chromatography using 80/20 to 100/0 dichloromethane/hexanes to afford 54f.

[00497] The desired product 54g was obtained by the procedure described for Example 26 Step B using 54d and 54f as the starting materials. The reaction was carried out in chloroform at 50° C. The residue was purified by flash chromatography using 85/15 hexanes/EtOAc to provide 54g in 56% yield. HRMS (FAB) Calcd for C₂₅H₃₃N₂O₄: 425.2440 (M+H)⁺. Found: 425.2424.

[00498] The desired compound 54h was obtained by the procedure described earlier for Example 1 Step I using EtOH as the solvent.

[00499] The desired compound 54i was prepared by the protocol described for Example 18 Step B.

[00500] The desired product 54j was obtained by the procedure described for Example 1 Step C. The crude material was used without purification.

[00501] The desired product 54k was obtained by the coupling protocol described previously for Example 1 Step D.

[00502] The desired product 54l is obtained by the hydrogenation procedure described previously for Example 1 Step G.

[00503] The desired product 54m is obtained by the cyclization protocol described for Example 1 Step H.

[00504] The desired product 54n is obtained by the producer described previously for Example 3 Step A.

[00505] The expected product 54o is synthesized as described earlier for Example 1 Step J.

Example 55

Preparation of Compound 55

[00507]

[00508] Commercially available 55a was converted to the desired product 55b using the procedure described for Example 26 Step A in 41% yield.

[00509] The desired product 55c was obtained by the coupling protocol described previously for Example 1 Step D using N-boc-t-butylglycine as the coupling partner. Purification using 95/5 dichloromethane/EtOAc afforded 55c in 57% yield.

[00510] The desired product 55d was obtained by the procedure described previously for Example 1 Step C. The crude material was carried further.

[00511] The desired product 55e was obtained by the protocol described for Example 26 Step B. Purification using 80/20 dichloromethane/EtOAc afforded 55e in 20% yield. HRMS (FAB) Calcd for C₃₂H₄₅N₂O₈: 585.3176 (M+H)⁺. Found: 585.3177.

[00512] The desired product 55f was obtained by the producer described previously for Example 1 Step G. HRMS (FAB) Calcd for C₂₅H₃₉N₂O₈: 495.2706 (M+H)⁺. Found: 495.2704.

[00513] The expected product 55g was synthesized as described earlier for Example 1 Step H. Purification by flash chromatography using 85/15 dichloromethane/EtOAc provided 55g in 10% yield.

[00514] The desired product 55h was obtained by the method described previously for Example 3 Step A. The crude material was carried further without purification.

[00515] The expected product 55i was synthesized as described earlier for Example 1 Step J. The crude material was carried further without purification.

Example 56

Preparation of Compound 56

[00517]

[00518] To a solution of commercially available methyl ester 10a (5.0 g, 20.4 mmol) in MeOH (20 mL) was added a solution of LiOH (730 mg, 30.6 mmol) in water (20 mL). The reaction mixture was stirred at ambient temperature for 2 hrs. TLC indicated consumption of starting material. The reaction mixture was concentrated and acidified with 10% citric acid solution. Solid NaCl was added and the aqueous layer was extracted with EtOAc several times. The combined EtOAc layer was dried (Na₂SO₄) and concentrated to provide 56a in quantitative yield. HRMS (FAB) Calcd for C₁₀H₁₈N₁O₅: 232.1185 (M+H)⁺. Found: 232.1189.]

[00519] The desired product 56b was obtained by the method described for Example 1, Step A. The crude material was converted to the methyl ester without purification.

[00520] The desired product 56c was obtained by the method described for Example 1, Step B. Purification of the residue by column chromatography using 80/20 to 50/50 hexanes/EtOAc and then 70/30 to 40/60 dichloromethane/EtOAc afforded 13% of 56c. HRMS (FAB) Calcd for C₂₁H₃₁NO₆: 394.2230 (M+H)⁺. Found: 394.2224.

[00521] The desired product 56d was obtained by the method described for Example 1, Step C. The crude material was used without purification.

[00522] The desired product 56e was obtained by the method described for Example 1, Step D, using N-boc-tert-butyl glycine as the coupling partner. Purification of the residue by column chromatography using 90/10 dichloromethane/EtOAc afforded 86% of 56e. HRMS (FAB) Calcd for C₂₇H₄₃N₂O₇: 507.3070 (M+H)⁺. Found: 507.3072.

[00523] The desired compound 56f was prepared by the protocol described for Example 1, Step E. The material was carried forward as it was.

[00524] The desired product 56g was obtained by the procedure described for Example 1, Step F. The material was purified by flash column chromatography using 98/2 dichloromethane/MeOH to yield 56g in 78% as a white foam. HRMS (FAB) Calcd for C₊H₄₁N₂O₇: 541.2914 (M+H)⁺. Found: 541.2916.

[00525] The desired product 56h was obtained by the procedure described for Example 1, Step G. The product obtained after filtering off the catalyst was pure enough for subsequent manipulations. HRMS (FAB) Calcd for C₂₃H₃₅N₂O₇: 451.2444 (M+H)⁺. Found: 451.2449.

[00526] The desired product 56i was obtained by the procedure described for Example 1, Step H. Purification of the crude residue using 75/25 hexanes/acetone provided a mixture of the product 56i along with triphenylphosphine oxide. HRMS (FAB) Calcd for C₂₃H₃₃N₂O₆: 433.2339 (M+H)⁺. Found: 433.2343.

[00527] The expected product 56j was synthesized as described for Example 1, Step I. Yield for two steps=16%. HRMS (FAB) Calcd for C₂₂H₃₁N₂O₆: 419.2182 (M+H)⁺. Found: 419.2176.

[00528] The expected product 56k was synthesized as described earlier for the Example 1, Step J. The material after work-up was of sufficient purity to be carried forward to the next step. HRMS (FAB) Calcd for C₄₀H₅₇N₆O₉: 765.4187 (M+H)⁺. Found: 765.4198.

Example 57

Preparation of Compounds of Formulas 57A and 57B

[00530]

[00531] To the solution of the commercial dihydrate of the amino acid (3S)-7-hydroxy-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid 57a (5.0 g, 21.8 mmol) in methanol (180 mL) was added concentrated hydrochloric acid (5.0 mL, 60 mmol). The resulting clear solution was then heated to reflux in an oil bath for 18 h. Solvents were removed in vacuo to give methyl ester 57b as a white solid, which was used in the next reaction without further purification.

[00532] To a solution of amine hydrochloride 57b, N-Boc-cyclohexylglycine (5.95 g, 21.8 mmol), HOOBt (3.73 g, 22.9 mmol) and EDCl (5.00 g, 26.1 mmol) in anhydrous DMF (200 mL) and CH₂Cl₂ (200 mL) at −20° C. was added NMM (7.20 mL, 65.5 mmol). After stirred at this temperature for 30 min, the reaction mixture was kept in a freezer overnight (18 h), after which EtOAc (600 mL), brine (150 mL) and 5% H₃PO₄ (150 mL) were added. The separated organic solution was washed with 5% H₃PO₄ (200 mL), saturated aqueous sodium bicarbonate solution (2×200 mL), water (200 mL), and brine (200 mL), dried with magnesium sulfate, filtered and concentrated in vacuo to afford 57c (10.3 g, quant. 2 steps) as a white solid. ¹H NMR (400 MHz, d₆-DMSO) δ 9.32 (s, 1H), 8.35-8.32 (m, 1H), 6.99 (d, J=8.3 Hz, 1H), 6.65-6.54 (m, 1H), 4.92 (d, J=15.5 Hz, 1H), 4.50 (d, J=15.5 Hz, 1H), 4.43-4.36 (m, 1H), 4.29-4.19 (m, 1H), 3.53 (s, 3H), 3.02-2.81 (m, 2H), 1.98-1.62 (m, 8H), 1.42-1.11 (m, 14H); ¹³C NMR (100 MHz, d₆-DMSO) δ 171.6, 171.2, 162.3, 156.0, 133.7, 128.8, 125.3, 114.1, 112.6, 78.0, 54.8, 52.4, 51.9, 45.0, 29.5, 28.1, 28.0, 25.9, 25.6, 25.5; HRMS m/z 447.2492 [calcd for C₂₄H₃₄N₂O₆, 447.2495].

[00533] The Boc-amino methyl ester 57c (7.20 g, 16.1 mmol) was dissolved in 4 N HCl (100 mL, 400 mmol) and the resulting solution was stirred at rt. The progress of the reaction was monitored by TLC. After 4 h, the solution was concentrated in vacuo and the residue was under vacuum overnight to give 57d as a white solid which was used in the next coupling reaction without further purification.

[00534] To a solution of amine hydrochloride 57d (from Step D), 6-heptenoic acid (2.90 g, 22.6 mmol), HOOBT (3.70 g, 22.7 mmol) and EDCl (4.80 g, 25.0 mmol) in anhydrous DMF (250 mL) and CH₂Cl₂ (150 mL) at −20° C. was added NMM (7.50 mL, 68.2 mmol). After stirred at this temperature for 30 min, the reaction mixture was kept in a freezer for 2 days. It was then stirred in air and allowed to warm to rt. in 1 h. EtOAc (500 mL), brine (100 mL) and 5% H₃PO₄ (100 mL) were added. The separated organic solution was washed with 5% H₃PO₄ (100 mL), saturated aqueous sodium bicarbonate solution (2×150 mL), water (150 mL), and brine (150 mL), dried with magnesium sulfate, filtered and concentrated in vacuo. Flash chromatography (5 to 30% EtOAc-Cl₂Cl₂) afforded 57e (2.30 g, 5.04 mmol, 31% (2 steps)) as a white solid. ¹H NMR (400 MHz, d₆-DMSO) δ 9.32 (s, 1H), 8.06-8.01 (m, 1H), 7.00-6.6.96 (m, 1H), 6.63-6.54 (m, 2H), 5.78-5.70 (m, 1H), 5.04-4.89 (m, 4H), 4.73-4.69 (m, 1H), 4.53 (d, J=15.5 Hz, 1H), 3.54 (s, 3H), 3.01-2.91 (m, 2H), 2.15-1.93 (m, 4H), 1.76-0.97 (m, 15H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.0, 171.5, 171.1, 156.0, 138.6, 134.0, 128.7, 122.5, 114.6, 114.1, 112.5, 52.9, 52.2, 51.9, 45.0, 34.5, 32.8, 32.7, 29.4, 28.7, 28.1, 27.7, 25.9, 25.5, 25.5, 24.8; HRMS m/z 457.1 [calcd for C₂₆H₃₆N₂O₅, 456.6].

[00535] To the solution of 57e (2.20 g, 4.82 mmol) in anhydrous THF (100 mL) under nitrogen at 0° C. was added borane-THF solution (20 mL, 1.0 M, 20 mmol) cautiously. The resulting solution was stirred at 0° C. under hydrogen for 1 h 40 min. Then ethanol (10 mL) and pH 7 buffer (15 mL) were added, followed by 30% H₂O₂ solution (15 mL). After stirred at 0° C. for 20 min, it was warmed to rt. and stirred for 2 h. EtOAc (400 mL) and brine (200 mL) were added and layers were separated. Aqueous solution was extracted with EtOAc (2×150 mL). Combined organic solution was dried with magnesium sulfate, filtrated, concentrated in vacuo. Flash chromatography (3 to 5% MeOH—Cl₂Cl₂) afforded 57f (2.18 g, 4.47 mmol, 93%) as a white solid. ¹H NMR (400 MHz, d₆-DMSO) δ 9.32 (s,1H), 8.04-8.00 (m, 1H), 6.99-6.96 (m, 1H), 6.63-6.51 (m, 2H), 5.05-5.00 (m, 1H), 4.73-4.21 (m, 3H), 4.51 (d, J=15.5 Hz, 1H), 3.54 (s, 3H), 3.03-2.90 (m, 2H), 2.15-2.00 (m, 2H), 1.75-1.56 (m, 6H), 1.49-0.97 (m, 13H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.2, 171.6, 171.2, 156.0, 134.1, 128.7, 122.6, 114.2, 112.5, 60.7, 52.9, 52.3, 51.9, 45.1, 34.8, 32.4, 29.5, 28.7, 28.53, 28.47, 28.10, 25.9, 25.6, 25.5, 25.4, 25.2; HRMS m/z 475.2812 [calcd for C₂₆H₃₈N₂O₆, 475.2808].

[00536] A solution of phenol alcohol 57f (2.08 g, 4.38 mmol) and ADDP (3.00 g, 11.9 mmol) in anhydrous CH₂Cl₂ was bubbled with Argon through a frit glass bubbler for 20 min. To this solution at 0° C. was added triphenylphosphine (3.45 g, 13.2 mmol). After stirring at 0° C. for 20 min, the solution was warmed to rt. and stirred overnight (18 h) under nitrogen. TLC indicated the presence of substantial amount of starting material. A second batch of ADDP (3.00 g, 11.9 mmol) and triphenylphospine (3.45 g, 13.2 mmol) were added, and the mixture was stirred under nitrogen for 2 days and 16 h. TLC showed the complete consumption of the starting material. After removal of solvent in vacuo, the residue was partially purified by flash chromatography (1 to 2% MeOH in Cl₂Cl₂) to afford a mixture of the macrocycle 57 g and triphenylphosphine oxide. The macrocyclic 57 g was hydrolyzed to the corresponding acid without further purification. ¹H NMR (400 MHz, d₆-DMSO) δ 8.00 (d, J=10.0 Hz, 1H), 7.17 (d, J=8.2 Hz, 1H), 6.82-6.76 (m, 2H), 5.14 (d, J=14.5 Hz, 1H), 4.79-4.74 (m, 1H), 4.39 (dd, J=11.5, 6.4 Hz, 1H), 4.25 (d, J=14.7 Hz, 1H), 4.22-4.18 (m, 1H), 4.08-4.02 (m, 1H), 3.68 (s, 3H), 3.18 (dd, J=15.1, 6.4 Hz, 1H), 2.85 (dd, J=14.7, 11.5 Hz, 1H), 2.07-2.04 (m, 2H), 1.81-1.40 (m, 10H), 1.32-0.85 (m, 9H); ¹³C NMR (100 MHz, d₆-DMSO) δ 171.9, 171.5, 170.2, 157.1, 137.0, 131.5, 126.4, 115.9, 112.6, 66.5, 54.4, 52.2, 51.9, 46.8, 44.9, 44.4, 33.6, 29.4, 29.1, 28.0, 27.3, 27.0, 26.0, 25.3, 25.2, 24.3, 24.2, 23.9; HRMS m/z 457.2707 [calcd for C₂₆H₃₆N₂O₅, 457.2702].

[00537] An aqueous lithium hydroxide solution (0.21 g, 30 mL H₂O, 8.75 mmol) was added to a 0° C. solution of methyl ester 57 g (from step 12F) in THF (30 mL) and methanol (30 mL). The mixture was stirred in an ice bath and warmed to rt. along with it in 4 h. The progress of the reaction was monitored by TLC. After the volatiles were removed in vacuo, EtOAc (100 mL) and water (30 mL) were added and the two layers separated. The aqueous solution was extracted again with CH₂Cl₂ (100 mL), after which it was acidified to pH=1. EtOAc was then added (150 mL) and the aqueous solution was saturated with solid sodium chloride. After separation of the layers, the aqueous layer was extracted with EtOAc (2×100 mL). Organic solutions were combined, dried with magnesium sulfate, filtered and concentrated in vacuo to afford 57 h (1.23 g, 2.78 mmol, 63% (2 steps)) as a white solid.

[00538] To a solution of acid 57 h (0.390 g, 0.881 mmol), amine A (0.360 g, 0.898 mmol), HOOBt (160 mg, 0.981 mmol) and EDCl (210 mg, 1.10 mmol) in anhydrous DMF (50 mL) and CH₂Cl₂ (30 mL) at −20° C. was added NMM (0.40 mL, 3.64 mmol). After stirred at this temperature for 30 min, the reaction mixture was kept in a freezer for 66 h. Then EtOAc (200 mL), brine (50 mL) and 5% H₃PO₄ (50 mL) were added. The separated organic solution was washed, successively, with 5% H₃PO₄ (80 mL), saturated aqueous sodium bicarbonate solution (2×80 mL), water (80 mL), and brine (80 mL), dried with magnesium sulfate, filtered and concentrated in vacuo. Flash chromatography (2 to 5% MeOH—Cl₂Cl₂) afforded 57i as a mixture of four diastereomers (0.340 g, 0.431 mmol, 49%) as a white solid. ¹H NMR (400 MHz, d₆-DMSO) δ 8.56-8.46 (m, 1H), 7.96-7.82 (m, 2H), 7.40-7.25 (m, 6H), 7.15-6.99 (m, 2 H), 6.81-6.74 (m, 2H), 6.05-5.71 (m, 2H), 5.11-5.02 (m, 1H), 4.85-4.68 (m, 1 H), 4.40-3.70 (m, 8H), 3.14-3.02 (m, 1H), 2.95-2.73 (m, 7H), 2.06-2.05 (m, 2 H), 1.81-1.39 (m, 10H), 1.30-1.05 (m, 11H), 0.89-0.75 (m, 5H); ¹³C NMR (100 MHz, d₆-DMSO) δ 172.24, 172.21, 171.87, 171.81, 171.78, 171.7, 170.8, 170.77, 170.74, 170.5, 170.4, 170.0, 169.97, 169.24, 169.22, 169.1, 169.0, 168.0, 167.9, 167.82, 167.78, 157.2, 156.9, 137.61, 137.57, 137.54, 137.47, 137.43, 137.38, 133.2, 132.2, 128.9, 128.44, 128.41, 128.37, 128.0, 127.96, 127.6, 127.4, 127.3, 127.19, 127.16, 115.7, 115.6, 115.5, 112.8, 112.77, 112.7, 112.6, 73.6, 73.39, 73.37, 72.4, 71.7, 66.9, 66.7, 55.8, 55.6, 55.09, 55.07, 53.02, 52.95, 52.9, 52.6, 51.0, 50.96, 50.91, 50.86, 50.76, 45.6, 45.5, 45.44, 45.36, 41.7, 41.6, 41.5, 41.4, 36.6, 36.55, 36.49, 35.3, 33.7, 33.6, 33.5, 33.0, 32.4, 30.7, 30.3, 30.1, 30.0, 29.8, 29.48, 29.45, 29.41, 28.3, 28.2, 28.1, 27.3, 27.2, 27.13, 27.09, 27.0, 26.9, 26.85, 26.82, 26.1, 25.4, 25.2, 24.1, 24.08, 24.03, 24.0, 23.9, 23.8, 18.8, 18.7, 18.6, 18.4, 13.9, 13.8, 13.7; HRMS m/z 789.4560 [calcd for C₄₃H₆₀N₆O₈, 789.4551, error=1 ppm].

Example 58

Preparation of Compound of Formula 58

[00540]

[00541] The desired compound 58a was prepared according to the method of Example 1, Step J, except substituting amine hydrochloride B for A. The hydroxy amide 58a was obtained as a mixture of inseparable diastereomers in the form of a white solid in 53% yield.

Example 59

Preparation of Compounds of Formulas 59A and 59B

[00543]

Example 60

Preparation of Compound 60

[00545]

[00546] A solution of [CpRu(η⁶-4-chlorophenylpropionic acid)]PF₆ (4.14 g, 8.36 mmol) in dry DMF (20 mL) was treated with HOBt (1.69 g, 12.54 mmol, 1.5 equiv.) and Hunigs base (6.47 g, 9.20 mL, 50.16 mmol, 6.0 equiv.) The reaction mixture was cooled to 0° C. and treated with EDCl (2.39 g, 12.54 mmol, 1.5 equiv.) The reaction mixture was stirred at 0° C. for 30 min and the Tic-ammonium salt 57d (2.90 g, 7.6 mmol mmol, 1.0 equiv.) was added. The reaction mixture was stirred at rt. for 12 h and the DMF was distilled out in vacuo. The residue was diluted with aq. HCl (1M, 100 mL) and extracted into CH₂Cl₂ (3×100 mL). The combined organic layers were washed with aq. NaHCO₃ (1×100 mL), brine (100 mL), dried (Na₂SO₄), filtered, concentrated in vacuo to yield a brown solid 60a (5.2 g, 83%) which was used for cyclization. MS: (Electron spray): 647 [(M-CH₃OH—PF₆)⁺, 100]. HRMS calcd. for C₃₂H₃₄ClN₂O₄Ru [(M-CH₃OH—PF₆)]⁺ 647.1256; Found: 647.1241.

[00548] The cyclized compound from the previous step was dissolved in CH₃CN (50 mL) and filtered into a quartz tube. The solution was degassed and photolysed in a Raynot instrument (λ=350 nm) for 48 h. The reaction mixture was concentrated in vacuo and the residue was purified by chromatography (SiO₂, EtOAc/Hexanes 3:2) to yield a tan colored solid 60b (289 mg, 20%). R_f: 0.73 (acetone/hexanes 3:7) ¹H NMR (CDCl₃, 300 MHz) δ 7.18 (d, 2H, J=8.1 Hz), 7.20-7.09 (m, 2H), 6.92 (d, 2H, J=7.8 Hz), 6.86 (dd, 1H, J=2.1, 7.2 Hz), 6.76 (s, 1H), 5.41 (d, 1H, J=17.4 Hz), 4.23-4.18 (m, 2H), 4.00 (bs, 1H), 3.68 (s, 3H), 3.41 (dd, 1 H J=12, 3.9 Hz), 3.01-2.86 (m, 1H), 1.9-1.62 (m, 4H), 1.52 (bd, 1H, J=9.3 Hz), 1.36-1.07 (m, 5H); ¹³C NMR (CDCl₃, 100 MHz, δ) 173.2, 167.2, 163.8, 156.5, 155.1, 135.8, 132.7, 130.2, 129.6, 126.2, 119.1, 117.5, 115.8, 60.2, 55.5, 51.6, 44.2, 42.0, 35.7, 30.2, 29.3, 26.5, 26.2, 25.8, 25.7 MS: (Electron spray): 477 [(M+1)⁺, 100], 315 (20); HRMS calcd. for C₂₈H₃₃N₂O₅ (M+1)⁺: 477.2389; Found 477.2375; CHN Calcd. for C₂₈H₃₂N₂O₅.0.5H₂O: C 69.26% H 6.85% N 5.77%; Found: C 69.62% H 6.59% N 5.77%

[00549] A solution of methyl ester 60b (235 mg, 0.5 mmol) of Tic-macrocycle in dioxane(10.0 mL), H₂O (10.0 mL), CH₃OH (50.0 mL) was treated with LiOH.H₂O (41 mg, 1.0 mmol, 2.0 equiv.) and stirred at rt. for 3 h. The reaction mixture was acidified and (4M HCl in Dioxane). The reaction mixture was concentrated in vacuo and the remaining water was frozen and lyophilized to obtain a colorless solid 60c used for coupling.

[00550] A solution of the hydrolyzed acid 60c (0.5 mmol) in dry DMF (5.0 mL) and CH₂Cl₂ (5.0 mL) was treated with HOOBt (132 mg, 0.75 mmol, 1.5 equiv.), and cooled to 0° C. and Hünigs base (258 mg, 2.0 mmol, 4.0 equiv., 369 μL) was added. To this mixture was added EDCl (143 mg, 0.75 mmol, 1.5 equiv.) and amine hydrochloride B (214 mg, 0.5 mmol, 1.0 equiv.) sequentially. The reaction mixture was stored in freezer for 48 h and concentrated in vacuo to remove DMF and CH₂Cl₂. The residue was diluted with aq. HCl (2M, 50 mL) and extracted with CH₂Cl₂ (3×30 mL) The combined organic layer was extracted with aq. HCl (1M, 2×50 mL), aq. NaOH (2M 2×30 mL), brine, dried (MgSO₄) and concentrated in vacuo. The residue 60d (172 mg) was oxidized without further purification; MS: (Electron spray): 838 [(M+1)⁺, 50], 490 (100); HRMS calcd. for C₄₇H₆₀N₅O₉ (M+1)⁺: 838.4391; Found: 838.4398.

Example 61

Preparation of Compound 61

[00552]

Example 62

Preparation of Compound 62

[00554]

Example 63

Preparation of Compound 63

[00556]

[00557] A solution of [CpRu(η⁶-4-chlorophenylpentanoic acid)]PF₆ (2.2 g, 4.0 mmol) in dry DMF (10 mL) was treated with HOBt (810 mg 5.99 mmol, 1.5 equiv.) and Hünigs base (2.58 g, 3.6 mL, 19.9 mmol, 5.0 equiv.) The reaction mixture was cooled to 0° C. and treated with EDCl (1.14 g, 6.0 mmol, 1.5 equiv.) The reaction mixture was stirred at 0° C. for 30 min. and the Tic-ammonium salt 57d (1.60 g, 4.0 mmol, 1.0 equiv.) was added. The reaction mixture was stirred at rt. for 12 and the DMF was distilled out in vacuo. The residue was diluted with aq. HCl (1M, 100 mL) and extracted into CH₂Cl₂ (3×100 mL). The combined organic layers were extracted with aq. NaHCO₃ (1×40 mL), brine (100 mL), dried (Na₂SO₄), filtered, concentrated in vacuo to yield a brown solid 63a (2.41 g, 75%) which was used for cyclization.

[00559] The cyclized compound from the previous step was dissolved in CH₃CN (60 mL) and filtered into a quartz tube. The solution was degassed and photolyzed in a Raynot (λ=350 nm) for 48 h. The reaction mixture was concentrated in vacuo and the residue was purified by chromatography (SiO₂, acetone/Hexanes 3:7) to yield a tan colored solid 63b (140 mg, 13%).; R_f: 0.73 (acetone/hexanes 3:7); MS: (FAB): 505 [(M+1)⁺, 80], 232 (40); HRMS calcd. for C₃₀H₃₇N₂O₅ (M+1)⁺: 505.2702; Found: 505.2698.

[00560] A solution of methyl ester 63b (235 mg, 0.5 mmol) of Tic-macrocycle in dioxane(10.0 mL), H₂O (10.0 mL), CH₃OH (50.0 mL) was treated with LiOH.H₂O (41 mg, 1.0 mmol, 2.0 equiv.) and stirred at rt. for 3 h. The reaction mixture was acidified and (4M HCl in Dioxane). The reaction mixture was concentrated in vacuo and the remaining water was frozen and lyophilized to obtain a colorless solid 63c used for coupling.

[00561] A solution of the hydrolyzed acid 63c (100 mg, 0.21. mmol) in dry DMF (4.0 mL) and CH₂Cl₂ (2.0 mL) was cooled to 0° C. and treated with HOOBt (53 mg, 0.32 mmol, 1.5 equiv.), Hunigs base (122 mg, 0.95 mmol, 4.5 equiv., 175 μL), EDCl (61.0 mg, 0.32 mmol, 1.5 equiv.) and stirred for 0.5 h and treated with the amine hydrochloride A (100 mg, 0.25 mmol, 1 equiv.). The reaction mixture was stirred at rt. for 16 h and concentrated in vacuo to remove DMF and CH₂Cl₂. The residue was diluted with aq. HCl (2M, 50 mL) and extracted with CH₂Cl₂ (3×50 mL) The combined organic layer was extracted with aq. HCl (1M, 100 mL), aq. NaOH (2M 100) brine, dried (Na₂SO₄) and concentrated in vacuo. The residue 63d (72 mg) was oxidized without further purification.

Example 64

Preparation of Compound 64

[00563]

[00564] The desired product 64a was obtained by the procedure described for Example 1, Step F. The material was purified by flash column chromatography using EtOAc/Hex (7:3) to yield 64a in 80%.; ¹H NMR (CDCl₃, δ): 7.35-7.29 (m, 5H), 7.02 (d, 2H, J=8.4 Hz), 6.72 (d, 2H, J=6.9Hz) 6.01 (d, 1H), 4.60 (t, 1H), 4.52 (s, 1H), 3.8-3.61 (m, 2H), 3.72 (s, 3 H), 3.54-3.51(m, 4H), 2.83 (t, 2H, J=7.5 Hz), 2.39 (t, 2H, J=8.1 Hz) 2.41-2.20 (m, 1H), 2.05-1.83 (m, 1H), 1.85-1.58 (m, 8H), 1.26-1.24 (m, 5H); ¹³C NMR (CDCl₃, δ): 172.2, 171.9, 171.0, 154.4, 138.3, 132.2, 129.4, 128.4, 127.7, 127.6, 115.4, 73.0, 66.9, 66.2, 57.9, 54.9, 52.5, 52.3, 41.0, 38.5, 34.7, 30.8, 30.0, 29.4, 27.9, 26.1, 26.0, 25.9.

[00565] The desired product 64b was obtained by the procedure described for Example 1, Step G. The product obtained after filtering off the catalyst was pure enough for next step.

[00566] The desired product 64c was obtained by the procedure described for Example 1, Step H. The crude reaction mixture was purified by SiO₂ gel chromatography (acetone/Hexanes 3:7) to yield 64c (64 mg, 16%) as a colorless solid.; ¹³C NMR (CDCl₃) ••172.1, 171.1, 171.0, 157.7, 131.0 129.9, 114.3, 78.1, 64.7, 63.3, 58.7, 55.3, 52.2, 52.0, 42.1, 37.9, 36.1, 30.8, 30.7, 29.7, 28.7, 28.5, 26.2, 26.0; MS (FAB) 473 (M+1)⁺, (100), 327 (20).

[00567] The acid was synthesized as described for Example 1, Step I in quantitative yield. The crude mixture after evaporation was directly used for the next step.

[00568] The expected product 64e was synthesized as described earlier for the Example 1, Step J. The coupled material was used directly for the next step to synthesize 64.

Example 65

Preparation of Compound 65

[00570]

Example 66

Preparation of Compounds of Formulas 66A and 66B

[00572]

Example 67

Preparation of Compounds of Formulas 67A and 67B

[00574]

Example 68

Preparation of Compound of Formula 68

[00576]

Example 69

Preparation of Compounds of Formula 69A and 69B

[00578]

Example 70

Preparation of Compounds of Formula 70A and 70B

[00580]

Example 71

Preparation of Compound of Formula 71

[00582]

Example 72

Preparation of Compound of Formula 72

[00584]

Example 73

Preparation of Compound of Formula 73

[00586]

Example 74:

Preparation of Compound 74A and 74B

[00588]

Example 75

Preparation of Compounds 75A and 75B

[00590]

Example 76

Preparation of Compound 76

[00592]

Example 77

Preparation of Compound 77

[00594]

Example 78

Preparation of Compound 78

[00596]

Example 79

Preparation of Compound 79

[00598]

Example 80

Preparation of Compound 80

[00600]

Example 81

Preparation of Compound 81

[00602]

Example 82

Preparation of Compound 82

[00604]

Example 83

Preparation of Compound 83

[00606]

Example 84

Preparation of Compound 84

[00608]

Example 85

Preparation of Compound 85

[00610]

Example 86

Preparation of Compound 86

[00612]

Example 87

Preparation of Compound 87

[00614]

Example 88

Preparation of Compound 88

[00616]

Example 89

Preparation of Compound 89

[00618]

Example 90

Preparation of Compound 90

[00620]

Example 91

Preparation of Compound 91

[00622]

Example 92

Preparation of Compound 92

[00624]

Example 93

Preparation of Compound 93

[00626]

Example 94

Preparation of Compound 94

[00628]

Example 95

Preparation of Compound 95

[00630]

Example 96

Preparation of Compound 96

[00632]

Example 97

Preparation of Compound 97

[00634]

Example 98

Preparation of Compound 98

[00636]

Example 99

Preparation of Compound 99

[00638]

Example 100

Preparation of Compound 100

[00640]

Example 101

Preparation of Compound 101

[00642]

Example 102

Preparation of Compound 102

[00644]

Example 103

Preparation of Compound 103

[00646]

Example 104

Preparation of Compound 104

[00648]

Example 105

Preparation of Compound 105

[00650]

Example 106

Preparation of Compound 106

[00652]

Example 107

Preparation of Compound 107

[00654]

Example 108

Preparation of Compound 108

[00656]

Example 109

Preparation of Compound 109

[00658]

Example 110

Preparation of Compound 110

[00660]

Example 111

Preparation of Compound 111

[00662]