SA00210014B1 - Macrocyclic peptides actvet the hepatitis c virus - Google Patents

Abstract

Abstract: The present invention relates to macrocyclic compounds of formula (I) that are active in in-vitro test tube and in cellular assays against the NS3 protease of hepatitis C virus (I), where D, R4, and R3 are A, R21, R22, w, and Ka are defined in this statement, or pharmaceutically acceptable salts, or ester, pharmaceutically acceptable salts.

Description

Macrocyclic peptides effective against hepatitis C virus Full description Scope of invention The present invention relates to “compounds” methods

Preparation of these compounds and methods for the treatment of hepatitis 6 virus (HCV) infection

hepatitis © virus (HCV) infection in particular; The present invention provides new Cla fin > peptide analogues; Pharmaceutical compositions contain

on these isotopes and ways to use these isotopes in the treatment of HOV infection

background of the invention

The hepatitis virus © (HOV) is the main causative agent of a type of hepatitis

Other than types C8 and 8 occur post-transfusion or community-acquired ‎community-acquired | 0 worldwide. It is estimated that more than 171 million people in

All over the world infected with the virus. A large percentage of carriers become infected

The virus carriers develop a chronically chronic form and many of them develop chronic liver disease

chronic liver disease This group, in turn, is vulnerable

Highly susceptible to serious liver disease such as cirrhosis, hepatocellular carcinoma, and terminal liver disease leading to death.

The mechanism by which hepatitis C virus remains active has not been clearly clarified

Causing a high rate of chronic liver disease. It is not known how 11017 interacts with

The host immune system and how to avoid it. In addition; Not even set

Now the roles of cellular and humoral immune responses

A v and the disease arising from that. HCV has been prescribed in the prevention of infection to prevent transfusion-associated viral hepatitis immunoglobulins immunoglobulins

Center for Disease Control However, “transfusion-associated viral hepatitis” does not currently recommend treatment with immunoglobulins for this purpose. And the lack of an appropriate response or preventive measures after the vaccine for the development of a protective and effective immune system impedes the development of this and in the short term; Hopes pin “post-exposure prophylaxis measures exposure.” Studies have been conducted entirely on antiviral interventions on various antiviral interventions with the aim of distinguishing pharmaceutical agents capable of treating infection in clinical studies in patients with hepatitis. Chronic© effectively. These studies included HCV with other combination antiviral agents in single form and in interferon-alpha combination using ye viruses. These studies have shown that a large number of participants did not respond to these habits, and it was found that a large percentage of those who responded satisfactorily relapsed after the end of treatment. Ctherapies is the only available treatment that has (IFN) interferon and until the past few years; .© has been a proven benefit clinically approved for the treatment of patients with chronic hepatitis Low treatment response rate 0 however long-term response rate Vo thyroiditis cretinopathy Causing serious side effects (ie Interferon B retinopathy negatively affects life (depression and depression) acute pancreatitis Acute pancreatitis: 5 approved for the treatment of ribavirin treated patients. Initially, the use of 10108800 was in combination with alone. But it It is now approved for the treatment of TPN patients who do not respond to treatment with B but HCV is in the gold standard treatment and is currently the gold standard untreated line Ye don't be afraid to use this combination treatment. IFN Accordingly, there are side effects caused by effective antiviral agents for the development of dala and overcome the limitations of current pharmaceutical therapies. HOV for the treatment of L. HOV infection

¢ and 1107 is an RNA virus (ribonucleic acid 63) enveloped positive strand RNA virus from the Flaviviridae family with a genome length of the RNA genome RNA A single-stranded HOV 4000 nucleotide Uy, & nucleotide is a single open reading frame (ORF) oo encoding a single large polyprotein aa) consisting of about 0 acid "-amino acid and in infected cells"; this polyprotein undergoes cleavage at multiple sites by cellular and viral proteases to produce NS -structural and non-structural proteins ( NS) proteins and, in the case of HOV, mature nonstructural proteins 01 (NS5B 5 ¢<NS5A «NS4B «<NS4A 3 «<NS2) mature nonstructural proteins are produced using two viral protease enzymes. Now, it performs the cleavage process at the junction between NS2 and 8183 and the second is a serine protease located within the N-terminal region of NS3 (hereinafter referred to as a protease). NS3 mediates all subsequent 65 cleavage steps after NS3 (adjacent to Vo n at the cleavage site in NS3-NS4A and opposite trans at the remaining sites in NSAB-NS5A < NSAA-NS4B and .NSSA It appears that NS4A protein (53%) has several functions as it may act as a cofactor for protease 3 and may participate in membrane localization of 1183 and other viral replicase components. Formation of a complex of the 1153 NSAAs© proteases appears to be necessary for the processing states to enhance the proteolytic efficiency of Jal proteins at all sites. The NS3 protein also shows the activity of a nucleoside triphosphatase and the -RNA helicase RNA and 08 is an RNA-dependent RNA polymerase involved in the replication of the hepatitis C virus

International Patent Application No. 4 47/0580 describes the enantiomer (=) of the nucleoside analogue of cytosine-1,3-oxathiolane (also known as 3TC) as an effective agent against HCV and with Previous clinical trials showed that this compound was Gal when used to treat HIV (human immunodeficiency virus) and 1187 (hepatitis B virus B), but it has not yet been clinically proven to be effective. To treat HOV, no ale mechanism against the virus has been mentioned yet.A general method for developing antiviral agents is to inactivate virally ancoded enzymes necessary for viral replication.In this regard, intensive efforts have led to the discovery of compounds that inhibit the NS3 protease. or helicase RNA 01 for HCV to the following: US Patent No. 0,177,784 described carboxamides bearing heterocyclic-substituted carboxamides and their analogues as effective agents against JHCV and directed these compounds toward helicase activity. 1153 protein in the virus, but no clinical trials have reported this yet, Chu and collaborators reported (in Tet.

Lott. p. 11-1717794 Vo 7m) that 01602010000006 is effective against HCV 1453 protease in the test tube. No further developments related to this compound were mentioned. Antiviral Research presented at the International Conference mentioned Ninth International Conference (Urabandi <Urabandi ¢Fukyshima, Japan 441M Research) +1 Vol. Thiazolidine is an inhibitor of HCV proteases, and many studies mentioned Aafia compounds for other serine protease enzymes such as elastase from human leukocyte elastase, and one family of these compounds was mentioned in the international patent application No. 98/777764 (Huchtist Marion Russell pf

:+ (Hoechst Marion Roussel 01 95 A.D.). The peptides mentioned in this application are morpholinylcarbonyl-benzoyl-peptide analogues structurally different from the peptides of the present invention. It disclosed International Patent Application No. 94/117674 from Vertex Pharmaceuticals° Inc. Vertex Pharmaceuticals Inc. reported on inhibitors of serine protease, specifically Hepatitis Virus Protease 3 ©. Hoffman LaRoche (see IPL No. 495 98/77; US Patents No. 9,477,744 and No. 1,0) also mentions hexapeptides as proteinase inhibitors useful as antiviral agents. To treat infection with 11017. Ve and Steinkuhler et al. and Ingallinella et al. describe the inhibition of the NS4A4B product (Journal Biochemistry vol a FY 844a-dtc, AVVE=AILT 1 (+) and patent application disclosure International No. 97/4770 issued by the Schering Corporation on peptide sequences consisting of 10 YY and yo amino acids that are effective against protease 1153 of HCV and reveal a patent application ISBN 475497/94 issued by Emory University on peptides and peptidomimetics effective in Cail testing against serine protease enzymes ISBN 47770/948 revealed About Ye Peptide Therapeutics Limited About a substrate of a depsipeptide that inhibits protease 1153 of -HCV oF pal US Patent No. ©,A74,Yor revealed molecules RNA enzymatic RNA molecules inhibit protease 1153 p1107.

SNP Any of the previous patent applications described above for proposed cyclic peptides that are active and selective against the 1153 protease of hepatitis virus 0. IPOs disclosed AQ VYYY 6 nos. 1/567. and 1/858 reported analogues of tetra- to hexapeptide- and tripeptide-binding 0-protease 1183. However, these discoveries did not suggest or lead to the design of the macrocyclic analogs to which the present invention relates. International Patent Application No. 388848/19 of August 0, 21994 by the Institute de Richerche di Biological Moleculare (IRBM) disclosed small peptides inhibitor of protease ©1153 to HOV and this disclosure is devoid of any Proposal or reference to the cyclic nature of the peptides of the present invention. In addition; This PCT application Gy was issued after the priority date of the present application. The international patent application No. 19/714467 issued by (RBM) after the priority date of this application also revealed short chain peptides oligopeptides with \o keto acids at Pl and the international patent application No. 44/560770 of Vertex Pharmaceuticals (issued October 7, 21999) also after the priority date of the present application. However, the Liles application does not refer to any cyclic peptides to which the present invention relates. One of the advantages of the present invention is that Supply of bulky cyclic protease inhibitory peptides © | 2183 of Cas infection virus Another advantage of one aspect of the present invention lies in the fact that these peptides specifically inhibit the NS3 protease and do not show significant inhibitory activity against enzymes other than other serine proteases Sie elastase from ¢human leukocyte elastase (HLE).

A

Bovine cporcine pancreatic elastase (PPE)

B Jia cysteine proteases or cysteine protease enzymes cbovine pancreatic chymotrypsin -human liver cathepsin B (Cat B) from human liver Another advantage of the present invention is the supply of small molecular weight peptides and the inactivation of cell membranes Low Able to penetrate cell membranes, molecular weight ° cell culture in cell culture, 1453 lis Another advantage of the compounds of the present invention also lies in the fact that they are effective in both types of what (B1 and A1) the main clinical isolates present In clinical isolates of genetic genotypes, it is highly indicated that these compounds will be effective against all known genotypes.

Hev currently nb 0 General description of the invention:)( The scope of the invention includes compounds of the formula

RA W, RZ > 0 :

R? JZ0

N: 9 5 4 3 8 1 LA

A

R

7 Name Se

R¢ for a q

where W stands for CH or for R* less alkyl ¢halo ¢H m cycloalkyl mo haloalkyl m “Cs. cycloalkoxy “Cys alkoxy” hydroxy or R® Cus (N(R), representing ten alkyl units of cycloalkyl sf as Mo; and

¢H J—— fay R? ° tweezers haloalkyl «Cs cycloalkyl «Cys alkyl m thioalkyl flexible» alkoxy port cycloalkoxy min” alkoxyalkyl bon Cs aryl “Cs, cycloalkyl or aryl from or Het Cus Het Represents a saturated or unsaturated heterocycle consisting of five, six or seven members and has from one to four heteroatoms chosen from oxygen «nitrogen ¢sulfur g

R¥ is substituted for Het or aryl <cycloalkyl and bears 01 «NO, «Cj cycloalkoxy morph alkoxy min» cycloalkyl m alkyl ¢halo 1 where T is less than (NH-C (O)-NH-R® 4 «NH-C(O)-R® is a lung of Mi; cycloalkyl of Cgalkyl separately represents each R® where each ¢Cy.¢ cycloalkyl of is reduced tola From R® where (NH-C(0)-OR%® a “© is reduced).

Vo 3 represents an NH, chydroxy or group of the form (NH-R¥ where R¥ is a Cg aryl transition aryl from heteroaryl "-C(0)-OR* R “-C(O)-NHR® where R? is an alkyl of §¢C, cycloalkyl D is a saturated or unsaturated alkylene chain of © to 01 atoms May contain one to three heteroatoms separately chosen from ©0;¢S or Cua (N-R*)

0 kj represents the alkyl <H of cycloalkyl min or R? Cua -C(0)-R*® is the alkyl of cycloalkyl min or Cg aryl or aryl©; and where the atoms of the D chain that form (ate ja the macrocyclic ring in structural formula (1) are numbered from left to right in structural formula 0) Tela ad from position AA)

thousand ye

D in the carbon chain has one to three substituents carried on any atom Shy or H Ba * «Cie alkyl mentioned; The aforementioned substituents shall be selected separately from the group consisting of and thioalkyl or thio maf ¢amino c¢halo ¢hydroxy from the alkoxy of haloalkyl “Crs alkyl” to be chosen from the group consisting of: R® where “-C(0)-NH-R® in the form amide represents A range; “carboxylic acid represents A or . Including, pharmaceutically acceptable Waa that contains an effective amount against GS 5 virus infection. The scope of this invention includes ester sl therapeutically acceptable salt J sia © liver from a compound with the formula ]; salt or carrier medium thereof; Mixed with a therapeutically acceptable carrier medium 0 Pharmacologically acceptable auxiliary agent An auxiliary agent in C An important aspect of the invention includes a method for treating hepatitis virus infection by giving the breast an effective amount against hepatitis virus © of the compound in formula 1 or mammalian mammals thereof or a composition as described above. “I” of hepatitis C virus © from compound of formula NS3 to a protease inhibitor amount or composition as described above. hepatitis virus© from a combination of the compound with the formula ]; Or salt or Tae by giving him some of it. According to one embodiment; The pharmaceutical compositions according to this invention include Ladle Accepted ester > © Additional. Examples of immunomodulatory agents include but are not limited to additional immunomodulatory agent A-0, B-and 8-interferons according to pharmaceutical compositions for an alternative embodiment; Pharmaceutical compositions may include Gig AA

1 00«121«H. Examples of antiviral agent of the present invention also include an antiviral agent amantadine and ribavirin for another alternative embodiment. Pharmaceutical compositions according to the present invention may also include iy.

HCV Other protease inhibitors for another alternative embodiment as well; Pharmaceutical compositions according to this invention may also include Gig ° as other HCV life cycle helicases HCV in the life cycle of targets

RES or metalloprotease cpolymerase chelicase polymerase Detailed description of preferred embodiments Definitions The following definitions apply unless otherwise indicated: lad) as used in this Ve using the number D to mean The term used in this statement to designate a position within the series at position Jia 0 of said 0 series” or “has series 01 of said Sia aaa gall (numbers)” or “has D series of said series 14 (VF at position double bond, “A” or similar term; VY 011 position or positions on a single double bond at position D

D When the atoms of the chain are numbered 0 as mentioned previously, ie; The D chain atoms within the vo chain that forms part of the massive ring in structural formula (1) are numbered from left to right in the formula

A from position no. Teli (1) structural or (9) to indicate the absolute formation (R) and by reference to cases where the sign is used in the context of oI) in the compound of the form R* Jie The position of the P2 PI acid moiety as used in this statement and the symbols indicate the C-terminus analogs of the peptide starting at the carboxy terminus amino acid residues amino from terminal 1 towards amino terminal 1760005 (ie 21 refers to position Talia AA

1

carboxylate and P2 denotes the second position of the carboxy terminus; ... etc.) (See what ela on Berger A. and Schechter I. in Transactions of the Royal ¢Society London vol. 3257 p. 154-749 010am) ..

The term “I-aminocyclopropyl-carboxylic acid (ACCA)” as used herein indicates

0 indicates a compound of the form: 0 perfect 03 and the term “vinyl-ACCA” as used herein | The statement refers to a compound of the form: 0 HoN.

The term “halo” as used herein means a substitute for halogen chosen from <bromo fluoro “chloro” or 1000

The term “haloalkyl M” as used herein means ‘either alone or in combination with

VY

It has a branched chain atom or a straight acyclic alkyl atom substituted for other substitutions of one or more halogens chosen with hydrogen so that one carbon atom is replaced by .iodo atoms or fluoro chloro < bromo as used in this statement; either alone or in combination with “© thivalkyl! the term means as a straight or branched-chain acyclic thiol containing another m-substituted alkyl group of .thiopropyl substituents as used herein; either” ( loweralkyl "or "© alkyl" and the term means a straight or branched chain of an acyclic alkyl atom alone or in combination with another substituent of <hexyl «butyl »propyl cethyl emethyl and includes carbon 1 to 1 atoms of -11-dimethylethyl and 2-methylpropyl “<1-methylpropyl ¢<l-methylethyl 01” as used herein; either alone or in combination with “Cycycloalkyl” meaning the term “ cyclopropyl carbon includes from three to six cycloalkyl atoms another substituent of cyclohexyl cyclopentyl unsaturated “cyclobutyl: cyclohexenyl” as the substituent cycloalkyl “and includes the term

ON

Saturated or unsaturated "as used in this section alkylene" and the term means one of hydrogen derived by removing a divalent divalent alkyl atom from a saturated or non-aliphatic Shu hydrocarbon each one of the ends of the saturated straight or branched chain and included; For example, -CH,CH,CH=CHCH,CH,- “-CH,CH,C(CHj;),CH,CH,- the noun” Y. No

VE

(For example: this oxygen contains a hetero-atom such as an alkyl and may contain a chain (CH,CH,-0-CH,-©) as used in this statement; either alone or in an alkoxy The term, as defined above, means that it contains what is not an alkyl, where -0-© alkyl is a combination with another substituent. The substituent is “1-methylethoxy” propoxy ¢ethoxy “methoxy ¢alkoxy” containing carbon atoms. 1 is more than 1 ° tert-butoxy The latter substituent is usually known as 1,1-dimethylethoxy and butoxy as used in this statement either alone or in combination “Cy cycloalkoxy” The term means © -0- contains from *?to + dd atoms. cycloalkyl from Sun with another substituent; from Sa as used in this statement; “© alkoxyalkyl” meaning the term

CNT as defined above and containing no more than an alkyl molar H©-0-o-OH, where 0 -.CH,-O-CH; B, methoxy methyl “and strengthened it. For example, beige is referred to as used in this statement either by itself or in acyl! The term means

Jie carbonyl group 1191© , related via the SAT group in combination with the -C(0)-C,6 alkyl substituent as used in this statement; either alone or in “©, aryl” or “©, aryl” * meaning the term Vo 1 contains an aromatic monocyclic system either a monocyclic aromatic system “Al” in combination with a substituent of 01 atoms containing aromatic bicyclic system or carbon atoms naphthyl or phenyl of a ring system eg an aryl ring system and includes carbon as used herein; either alone or in combination with “Coy aralkyl” the term as defined above means alkyl Cua alkyl de gana as defined above linked by another substituent aryl; x.-butylphenyl and benzyl eg caralkyl and includes carbon has from 1 to 7 atoms as used in this statement; either alone or in combination with another substitute "Het! the term means of a mixed saturated or unsaturated ring (including monovalent hydrogen derived by removing that aromatic Sua yo atom) consisting of five; Six or seven members with from one to four heterodimers, choose from sulfur 5 oxygen ¢nitrogen. Examples of suitable mixed rings are: tetrahydrofuran pyrimidine morpholine “dioxane” piperidine “isoxazole” diazepine <thiophene or the term includes "Het" is also a scrambled loop as defined above combined with one or more other loops which may be a scrambled loop or any other loop. One such example is thiazolo[4,5-b]-pyridine. Although the term "heteroaryl" falls within the definition of the term ale "Het", as used in this statement it means specifically an unsaturated mixed ring representing a system Lhe aromatic system Examples of suitable heteroaromatic systems are: ¢pyridine ¢indole ¢quinoline 01 N. 7 07 0 05

N didn't; Term with other substituent esters Compound of formula 1 Cus Replace any of the 'alkoxycarbonyl' functional groups with a ccarboxy functional group in molecule 0; the carboxyl terminal Vo a is preferred

0 is

where R&A selects in ester of an alkyl (eg “t-butyl “n-propyl cethyl methyl” Jal alkoxyalkyl ¢(n-butyl (eg “le) alkoxyacyl ¢(methoxymethyl) eg Jel le) aralkyl ¢ (acetoxymethyl eg; le) aryloxyalkyl ¢ (benzyl eg aryl ¢ (phenoxymethyl ° (eg phenyl) may carry Sud from alkyl chalogen ber or esters dy ©. other alkoxy suitable as prodrugs 885 in GUS Design of prodrugs by H. Bungaard (Bundgaard, H. Elsevier 6a) AAO (Elsevier) cited in This statement is for reference as a reference.These pharmaceutically acceptable esters are usually hydrolysed in the body

1. Of the compound in the acid form when injected into my breast and it turns into the acid form al

Lad 1 relates to the esters described above; It is recommended that any alkyl moiety present contain 1 to VU a carbon atom, specifically 1 to > atoms: HA unless otherwise stated. It is desirable that any aryl moiety present in such esters includes a .phenyl group

In particular the esters may be a benzyl ester «Cris alkyl ester bearing no substituents or a benzyl ester bearing at least one nitro «Cy, alkoxy»Cy alkyl halogen substituent or

.trifluoromethyl yo

The term "pharmaceutically acceptable salt" as used in this statement includes those salts derived from pharmaceutically acceptable bases. Examples of suitable bases include choline, ethanolamine, and ethylenediamine. The scope of the invention also includes the salts of nutella and Ca” (see also what sla on SM Berge 5.14.14 and co-workers in a paper titled Pharmaceutical salts

© in oJ, Pharm.

Sci. vol. 77 p. AVY 11-1 (2 cited in this statement for reference).

Preferred embodiments: :R' Preferred embodiments G5 of the present invention include compounds of Formula 1 as described above; Cua !8 is chosen from two different classes of stereoisomers diasteresisomers where the central 1-carbon atom has a configuration WSR is represented by the following two structural formulas (1) 5 (ii) : : 1 rT $ 8 1 , H 1 H . N N, : Ny IRA Nr IR A > 0 - 0 D D 0 adjoins amide in formula (:); or © adjoins m in formula (ii) and preferably that the linker chain D D is attached to R' in the adjacent configuration nm as represented by the structural formula (ii) :R? ye Preferred embodiments Gy of the present invention include compounds of formula 1 as described above; R? represents: RA W, RB = 2 where W preferably represents N Vo and preferably 2 represents the alkoxy «Cis alkyl ¢<H mf chloro ¢hydroxy or N( R®), where YYVY

a

2 Preferably H or alkyl represents m©. better; RY stands for H or alkoxy m©. More specifically, 12 represents .methoxy

preferably; R? represents a thioalkyl (H) alkoxy moiety of” phenyl or Het chosen from the group consisting of:

N N. 2c m - 3 1 ca 7 ¢ — ¢ \ S . 2 m £ with x Cr C0 has N\ / x N N—N l \ J ¢ and 2 ¢ N ¢ N RM ¢ 2 J 12 “mah mr 1 1 \ ed A N ¢ NN 0 4 24 J . preferably; (H day R?* alkyl group with NH- gl ¢NH-C(O)-R*® (NH-R® .NH-C(0)-OR® 4 C(O) -NH-R*® \. Better yet » 827 represents the M© alkoxy » phenyl or Het choose from the group consisting of:

V4 a R# N N:

A msq Te 1¢ l ¢ ser ¢ \ 5

RA

R#

N & 47 1 maz x da N—N and al aa and 4 02 4c live: RT NH-C(0)-OR* R ¢ NH- C(O)-R* “NH-R*” Cys alkyl represents Tn R* and preferably choose from the group consisting of: Het or cethoxy and more preferably; 1822 represents °

N N

= load — mm m ?; and 5 ¢ 5

R24 4 A and in the form of NH-C(0)-OR* R (NH-C(0)-R* (NH-R®) “Suit” is less and more

Ce alkyl of H represents 12 Sass more and in the form of Cy cycloalkyl I see Cig alkyl represented separately: Each R® and preferably; Each Ve is a meh. alkyl representing R® from. And better;

Ye. Who is the alkyl R? Preferably; R': described above; LST The preferred embodiments of the present invention include compounds of the formula (NH-C(O)-NH-R* with the formula urea “<NH-C(0)-R *? in amide form 83 preferably Cua or <8#. More frequently carbamate in the form “082-(111-0)0. Better yet, 8 represents carbamate or © .carbamate 3 represents “Sadi minb. And better” “12 represents cycloalkyl sf than an alkyl and preferably “th8 represents or cyclobutyl linal sat R? Maine. More preferably cycloalkyl or “ Crs alkyl .cyclopentyl :D ze where is the series of The preferred embodiments according to the present invention include compounds of formula

Jia atoms. It is better that A is saturated or unsaturated with 6 to alkylene bonding 0 chain atoms. 1 bonding chain 0 chain having on one or two hetero-atoms choose from: 0; 58 ;D and preferably; The chain optionally contains a hetero-atom D, and it is better for the chain to contain .7-©,.7 acyl Allah J©-A1 or NH 0 of 01 located at atom No. (N(AC) padi The most (N-C,; acyl or (NH) one to choose from: saturated. nitrogen chain. Most preferably the chain containing an atom, preferably S. Alternatively; containing chain 0 On one hetero-atom choose from: 0 or atoms that the © atom or 5 is located at position number 9 of the chain. 17 0 the length of the chain is Laie

R* and better”; .©,. alkyl or H representing R* where (RY) this series is preferably substituted by © and more preferably 8-(S)-Me 148-(5)-4 representing 11 or R * Spain or Wetness". More H Jia than that; the 0 chain has only one Yang double bond and the 0 chain is saturated. Preferably this double bond is in opposite position. VY and 11 places a

saturated or unsaturated all consisting of D chain alkylene atoms thereof; It represents a saturated Yau .carbon and its length is 7 D atoms. In this case; It is preferable that the -carbon chain be of the parent ¢hydroxy «thio coxo ¢H with R* Cua R* and it is better to carry the chain 0 instead of

JH JAR Sais ©. more alkyl or H Jia RY (Jail) and in the form of alkyl or alkoxy 10<(S)-Me or H representing R* and more preferably methyl© and containing in the form of carbon consisting of All of them are from alkylene atoms of a D chain that represents “ld of Yau” and it is better that this bond be carbon atoms 1 is preferred over one double bond and its length is that this double bond is in Sais of the chain. And the most VE and 1” binary between the two positions hydroxy d ¢H is less than R* where R* is an adjacent position. It is preferable to carry the chain ( instead of

JH Jha RY lI ©. Better yet, alkyl or H stands for RY or talc. Better yet, “alkoxy 1 10-(S)-Me” or “H” represents R* 1 obedience. More preferably tA The preferred embodiments according to the present invention include compounds of formula 1 [as described above; favorite represents atom 67); Represents 3 ¢C,¢ cycloalkyl ben or alkyl Represents: R® Cua Ar configuration saturated or unsaturated with 7 to 48 atoms attached to an alkylene representing a chain © | © Optionally containing one or two hetero-atoms selectable separately from: A adjacent to ¢N-C,; acyl a of which 24 is of SO or a hydroxy atom separately selectable from Jia to three fasts Shy represents 11; or RY

YIVY

YY

pharmaceutically acceptable ones. ester, salt, or “carboxylic acid” representing A as defined by R’. Preferred compounds are more compounds of formula 1 where ¢methoxy ol H is above; !2 represents Choose from the set consisting of: Het or “Cog alkoxy Jie RZ

N. > Fe —~ oy 5 ¢ 5 0 24d 2 © of

Cua

Ci ¢H stands for: R® where (NH-C(0)-NH-R* “(NH-C(O)-R* “NH-R* Mb alkyl H” C represents $C; cycloalkyl of ¢ alkyl $C, cycloalkyl sf of alkyl where "(NH-C(O)-OR*) or 'N' represents 0 and '2 represents H's atom; d is urea of the form NH-C(O)-NH-R¥? or a carbamate of the form (NH-C(0)-OR* where 13 alkyl Jia from cycloalkyl sl min; D represents a saturated or unsaturated alkylene chain containing 1 carbon atoms and 1 may contain 1 double bond between positions 11 and 11” or 11 and VE and may contain chain 0 mentioned on one hetero atom choose separately from: 0; 8; (NH ¢N(Ac) sl 1040 and cisalkyl .see H represents R* VY vy

R” and cmethoxy in compounds of formula 1 where 2 represents Sais and most compounds are ethoxy or ethoxy represents RM” —U where and $NH-C(0)- O-(Cy4 alkyl) or ¢NH-C(0)~(C, alkyl) <NH-(Cy4 alkyl) “Tes 11° represents a chain that is saturated or contains one double bond in adjacent position between The Two Positions ©

Ne is an actor in LS T Finally; The scope of the present invention includes all compounds in formula 4 to Tables 1 from comally. The pharmaceutical compositions according to this invention may be administered orally, preferably via an implanted reservoir or through an implanted reservoir. Parenterally, the intestinal contents of this invention contain any Gy carrier materials, and the pharmaceutical compositions may contain oral or parenteral injections.

Je and pharmaceutically acceptable non-toxic auxiliary agents or carriers and in some cases; The degree of -vehicles, adjuvants, auxiliary additives, or buffer solutions can be adjusted to the degree of bases, cacids, using formulation acids. Acidity of the formula | 0 To enhance the stability of the formed compound or to enhance the form used, Wana acceptable acidity buffers for its disposal. The term includes non-intestinal administration as used in this statement by cintracutaneous injection techniques into the skin Jala ¢subcutaneous injection or infusion or intra-articular intra-articular cintramuscular intra-articular muscle cintravenous vein intrathecal intrasternal sheath intrasternal cintrasynovial synovial cavity 7

YIVY ve intralesional site of infection, and the pharmaceutical compositions may be in the form of a sterile injectable preparation, a sterile aqueous or oily suspension, for example; As a suspension, sterile injectable preparation injectable. This suspension can be prepared according to techniques known in the technology using dispersing agents and Tween 80 agents (for example; wetting agents or wetting agents dispersing agents 0 are appropriate. suspending agents may be given as suspensions and pharmaceutical compositions pursuant to this invention by oral administration in any acceptable dosage form tablets capsules for oral administration including, but not limited to capsules and in the case of tablets -aqueous solutions, 501:0:5 aqueous suspensions and suspensions Aqueous ctablets .comn starch usually used carrier 1261056 and corn starch of gall used orally; include Ve and for administration of .magnesium stearate such as clubricating agents Jul ge lubricants also usually add starch useful lactose diluents for use diluted diluents of gall by oral route in the form of capsules; active ingredient includes dried maize. with specific emulsifying agents to taste. coloring and/or flavoring and/or flavoring vo and other suitable excipients or carriers for the preparation of the above-mentioned compositions and compositions can be found in standard pharmaceutical textbooks; For example, in the book Mac Publishing Company V4 edition (“Remington’s Pharmaceutical Sciences’ 1998 AD. (Pa.

Wild iy Easton State Easton <Mack Publishing Company mg/kg bw per day; and about 0.00 dose levels that fall between about © mg/kg bw per A day of YO inhibitor compounds preferably between about 0+ and about monotherapy one treatment de ja for the proteases described in this statement are useful for use in the form of pharmaceutical compositions are usually given according to this HOV for the prevention and treatment of the disease caused by vo is diluted to about © times daily or alternatively; the invention is given at a rate of about Ingredient Long or short term The amount of the active ingredient varies according to the treated host and the specific method of administration. (w/w) It is preferable that such preparations contain the active compound in an amount of

JA to approx 7700 from approx Cedric savvy in technology; Lower or higher doses than those for LS, described above, may be required. The dosage and specific treatment methods for any specific patient depend on various factors, the condition, body weight, body weight, cage, including the effectiveness of the particular compound used; age time of administration cdiet gender *""; Diet cgeneral health status general health severity drug combination of excretion rate of excretion ctime of administration of the patient for infection and the opinion of the attending physician. In general; asides panlly and injury progression of the peptide. optimum dose Treatment begins with small doses substantially below the optimum effect dose and thereafter; The dose is increased by small amounts until the optimal effect1 is achieved in these circumstances. general; It is highly desirable to give the compound at a concentration level that gives harmful or harmful side effects Effective results against viruses without causing any side effects Sale And when the compositions of this invention include a combination of a compound in the form!; one or more additional curative or prophylactic agents; Both the compound and the additional agent should be present at dose levels 7780 of the dose normally administered in 01 and best between about 00 to 0 lying between about 0 © pics of a single therapeutic dose. When these compounds are formed or its pharmaceutically acceptable salts with an acceptable carrier 1153 Pharmaceutical; The resulting structure may be given within a material body; like humans; to inhibit proteases

for 1107 or for the treatment or prevention of 1107 virus infection. Such treatment may also be achieved by using compounds of this invention in combination with agents including, but not limited to: immunomodulatory agents; such as or 7 interferons -8-"; Jul se other antivirals amantadine «ribavirin Jie other protease inhibitors 1193 I 1107; inhibitors of other target cells in the HCV life cycle such as the chelicase polymerase metalloprotease or internal ribosome entry site (IRES); or a combination thereof. Additional agents may be mixed with the compounds of this invention to form a single dose. Alternatively, these additional agents can be given to the breasts separately as part of a multiple dose. “lM Gig, another Gy embodiment of this invention provides Gola to inhibit the activity of the 1153 y.1 1507 protease in mammals by administering a compound of formula od wherein the substituents are as defined above. In a preferred embodiment; These methods are useful in reducing the activity of the HCV 1853 core protease in breasts. and if the pharmaceutical composition includes only a compound of this invention as the active ingredient, these methods may additionally include a step of administering to said mammal an agent chosen from an immunomodulatory agent; antiviral agent; An inhibitor of the HOV protease or an inhibitor of other Vo cells targeted in the HCV life cycle such as chelicase polymerase or metalloprotease This additional agent can be given to the mammal before; simultaneously with; or after giving the compositions of this invention. dg is an alternate preferred embodiment; These methods are useful for inhibiting viral replication in my breasts. Jia considers these methods useful in treating and preventing HOV. and if the dual. composition includes a compound according to this invention only as the active ingredient; These methods may additionally include a step of giving the said mammal an agent chosen from an immunomodulatory agent; antiviral agent; An inhibitor of the HOV protease or an inhibitor of other target cells in the HCV life cycle This additional agent may be administered to the mammal prior to; simultaneously with or after the administration of a

Composition Gd of this invention. The compounds mentioned in this statement may be used as laboratory reactants. reagents and the applicant provides for the first time low molecular weight, highly effective, specific compounds against proteases 1483 to 1107. Some of the compounds of the present invention may be useful in providing scientific research tools for the design of virus replication assays, validation of animal assay systems, and structural biological studies. structural biology studies to further advance knowledge of the mechanism of HCV disease. Compounds of the present invention may also be used to treat and prevent SgBl viral contamination of materials and thereby reduce the risk of viral infection to laboratory or medical personnel or patients who come into direct contact with such materials (for example PA 1;:; acyclic intermediates of compounds of Formula 1 in IPL Application No. 0/0904 and IPL Application No. 1/14 are cited herein for reference. The Gay compounds of the present invention are synthesized by the general process shown in Schematics II” I IIT (where PG represents appropriate protective groups). When the invention includes compounds of formula 1 A Cua representing (N-substituted amide homo-allyl ACCA (R') sf vinyl-ACCA is conjugated to an appropriate amine prior to its conjugation to P2 those skilled in the art will realize Coupling like that easily. And as those savvy in the tech will realize; circumnavigate

Ya

No such amide (A) is protected but does carry any suitable substituents of 85 as defined above. The ring-closing reaction (macrocyclization) takes place either by double olefin metathesis (see Scheme 1) or when the day chain contains a nitrogen atom by Cagis details these processes below: a. Massive ring formation by double exchange for olefin a

Ya 1 Chart PG or R? PG or R? PG or R? p or oo” or p or i H , b H , m0 HN. 0 N A — 0 N A

PG A PG / 8 O ° Ny 8 A, Hb 17 = 0 ِ a oH

H — a B

N N. A Hm N: 8 AH Core 7 N A 0 J 0 / 1 0 o 0 p - 0 ya - 0 588 except 0 a. D "ny D = : , "," A 71

Ib Ia Sn Saturated =D Unsaturated =D

N -_buffered carboxylic acid or amide with substituents at 4

Y zero or -

I planned through and perceivable coupling sequence There are several ways in which the coupling context can be performed The 4-(S)-hydroxyproline of the compound fess can be easily accomplished by the tech savvy. © (as) It is described in the Mitsunobu reaction by means of the 4-hydroxy substitution reaction of Mitsunobu by Rano »Tet. Lett. January 19481 CE; a r.

Krchnak Krchnak «Ter. Lett. Journal 10 944 (FVAY-TYVA) p. )-hydroxy bearing substituents of proline instead using the necessary assembly 1.4847 is carried out as described in the general processes in International Patent Application No. (see specific examples given below for these parts). 00/098588 and IPL No. 23 using P2 coupling techniques “Pl B; C: In short terms, the notches of the steps in IPL Bale Tau are described Known peptide coupling techniques. H.A. Black Miller, 8. 7. Same as that cited: 0 (SJ Miller Ru-based catalyst Ru 01 vol 7 Am. Chem. Soc. in Grubbs, R. H. and RH Gerbis Blackwell, HE. H Hoveda Bonitatebus, P.J. j. Bonitatibus cHarrity, 1. 7. A. Harriet M (—=) pp. 44-741 1994 CE and 71 Vol. Am. Chem. Soc. In Hoveyda, A H.

Nolan, 5. P. S. with me. Nsolan Stevens, E.D. Thee. d. Stephens Huang, J. J. Hwang eo “YE p YY vol oJ Am. Chem. Soc. In Petersen, J. 1. and J.L. Peterson will also realize that catalysts containing metals a) 499 7117 can be used for this reaction. Mo Jie Other transition metals a

A

(0

PCys

Cl,,,,

R=a”

PCy

Grubb Catalyst West BL 0

Qy,,

Ruse a

Poy, H

Hoveyda =) a 1 , Rus==x a”

PCys

Nolan catalyst optionally using double bond hydrogenation methods Step E: The carboxylic acid A' double bond is reduced in the technique. And when faa represents known standard hydrogenation methods ° . bond, the protective group is also removed from it appropriately (N containing Linkers b. Formation of a massive ring by a reductive amine (for the A-linker moieties)

When the binding moiety contains nitrogen Bd, the colossal ring is formed by the reductive amine as shown in Scheme 1 to obtain inhibitors with the general structural formula [1. Scheme II 2p R? R? / / / Qo Q A 2 A 2 A ,%8 | 8 N = a N hum N a .~ _ a ~ d ~ ME en = TE \ a 7 Ty . R? / 9 A Cx N 0 x “tx x x e carboxylic acid =A' buffer or amide bearing Soa of N “< a number from 1 to ©” - a number from 1 to e Step a: Addition of boron results in hydroboration of the ag double bond of Brown's procedure 01 (see ela About H.C. Brown © 1 and B.C. Subba Rao © 5, in “J.

Am.

Chem.

Soc. vol A) pp. 474 1477-76, 1904 AD) and then a vy

Oxidation of the resulting alcohol (le), for example, according to the Dess-Martin method in

Preparation 0600:0866 See what's in oJ Am.

Chem.

Soc. Volume OY p.: -1779797

corresponding aldehyde (21944), YYAY.

Step B: Hydrogenation in the presence of 2011 removes the protecting amino group © and then forms a massive ring by a reductive amine. and obtains the P3 unit

Used in this synthesis easily from a variety of glutamine, omnithine s lysine Jie, amino acids

(After the Jeli Hofmann Hofmann reaction, see what came in Ber magazine, vol. 946, p.: 7778,

other synthetic; These chemical modifications are considered to be amino acids and M1 ANY

modifications Taya's well-known methods of technology.

0 - Step C: The secondary amine ASH in the ©-linking moiety (formed after step d) is optionally done using alkyl halides or an acetyl group is added to it using alkyl acid chlorides or aryl acid chlorides using Known fas methodologies in the technique to obtain inhibitors with the general structural formula 11. When carboxylic acid A' represents a buffer, the protective group is removed from it as well.

10 c. Macrocyclization by forming a lactam

alternatively It is recognized that these bulky cyclic compounds with structural formulas 1 and 11 can be synthesized in other ways. for example; P1 and 23 can first be bound to bonding moiety 0, then coupled to 22 and the reaction of forming the massive ring may be lactam formation in two possible ways as those skilled in the technique will realize and as shown in the JIT diagram

p. 2® )1@ p 4 z OPG I' A : : IN A or N 8 A — 7 0 ora p worn itND R org © A 0 eset D 0 0 3p > 2c 1 4m( z N \ : D = 3c carboxylic acid =A' or amide substituent of N 8 zeros or Y ee = m number It ranges from 1 to 17 a vo

P1 Synthesis of tP1 Synthesis of inhibitors of general structural formulas 1 and 11 requires the same parts as 1.4847 in the international patent application vinyl-ACCA (a) Synthesis, dissolution, and joint pending applications are described by serial number /01 089888 and intl application no. 089888 is incorporated herein in its entirety for reference by reference) or 1/77/8855 M.) and 1 compound 0 (example homoallyl ACCA(b) synthesis P2 Certain parts of P2 used for the synthesis of compounds are described as Formula 1 in IPL Application No. 00/098547 and Application IPL No. 048988/00 and pending applications 0 shared by serial number 0/771,8775 The synthesis of other P2 fractions is incorporated into this statement in its entirety for reference. Het” carboxylic acid corresponding IVb Scheme IV RA NH i TO + Ae J. IVa ° vb RY —¢' . RZ N, RZ I | | Ht Ts Ve © vd of Vo

The synthesis is carried out according to the Jima procedure as reported by Li Li and his collaborators in the journal Med.Chem for vol VE p: 74097-754080, 144 a) and the intermediate compound TVa is prepared where OMe < 8 (Example; compound (of) as described by Brown et al. in oJ Med.

Chem. Vol. YY p.: Abalhakm, 84 Am. ° Step A: The intermediate compound IVb heterocyclic carboxylic acids — IVa is coupled under basic conditions using ,2001 to activate the carboxylate group and a variety of carboxylic acids with the general structural formula TVD are used to prepare the inhibitors; These carboxylic acids (las) may be available or synthesized as shown in Schematics 7, 71 and 11 or synthesized individually using the methods described in the particular examples. 1. Step b: The ring is closed “dormate”, then the water is dehydrated in Cay yb alkaline to obtain quinolines with the general structural formula IVA (i \) Synthesis of “Het”-carboxylic Acids With general structural formula IVb, the synthesis of amino-4-carboxy-aminothiazole derivatives bearing a substituent at site (Ve)1 uses a modified procedure described in ela by Berdikhina and collaborators in Chem.

Heterocycle. 42 Vo (English translation); Volume of p: 477-5477 AYA) 1 no

Chart 67 0 5 heat of A for H. + HO Br —> R® 0 Va Vb Ctg R Brilliant =

0Ve produces a variety of ¢2-alkylaminothiazolyl-d-carboxylic acids compounds with the general structural formula Ve using the general synthetic methodology shown in Scheme V using thioureas (of the formula (Va) with substituents and different (=R*) .3-bromopyruvic acid 5 (alkyl group) This type of condensation reaction is known as Tas in the technique. Alternatively, the P2 fraction containing thiazole derivatives is synthesized bearing an amino substituent at position 1 of the corresponding 2-carboxyl derivative as shown in Scheme VI according to the procedure described in © Unangst, P. About D.T. Connor 1 Connor, D. in “J Heterocyc.

Chem. 1 Volume YA Issue © p.: a) 47 0011-0117 .

Ya

VI Scheme 22 Na COOH oO B “m subject gene” A M1 45847 This process is in International Patent Application No. AB and is disclosed .. 0 /4688 and International Patent Application No. (VIId) ¢ at the site Shy carries the synthesis of 2-carboxy-aminothiazole derivatives ° compounds with the general structural formula ¢ 4-alkylthiazolyl-2-carboxylic acids producing a variety of ,

VIL using the general synthetic methodology shown in schematic 0 va

VII chart 5 0 - _omt char i wy oS 5) free 0 VIIa Vib

R# R#

N N

$= 0

EtO. 8 ho. 8 0 0

Vic VId a a a, and collaborators in Janusz uses the procedure described herein on Janusz 1194 CE with modifications described in YOYA-Y0NV0 Vol 541 p:7 Med. Chem. Different structural formula B-bromoketones with the following (Iv) ethyl thiooxamate: gail) ° reacts with the general structural formula thiazolyl carboxylic acids to form (alkyl group =R™) VIIb in general Tas condensation is known as Jeli and this type of VII (VIIb) carries 2-carboxy-imidazole substituents for the synthesis of derivative compounds with the general structural formula calkylimidazolyl-2-carboxylic acids producing a variety of . 7711 using the general synthetic methodology shown in Scheme 1706 Ve

VIII Scheme Rag Raw Tess Laba } N— 03 0

Villa Vib

Uses the procedure described in cla on Baird et al. in Amer.

Chem.

Soc. OYA vol. p: PAT, VETTE) M: The alkyl imidazole is deprotonated using a strong base (eg (nBuLi) and then reacts with CO, To form VITIb carboxylic acid and this type of condensation Jeli ° is known as jus in the old technique Synthesis of 4-hydroxy-7-methoxy-2-(imidazolyl or pyrazolyl)quinolines 4-Hydroxy-7-R* containing an imidazolyl or pyrazolyl moiety at C2 using the general synthetic methodology nd) in Scheme IX Scheme IX RY N, a R% N RZ x i x 0838 .083 Xa Xb

N, RZ 2 x ZZ OH Xc 1 =R? imidazole or pyrazole derivatives describe the synthesis of the intermediate compound | CP J (where (OMe = RA IXa 4-benzyloxy-2-chloro-7-methoxyquinoline) is detailed in example + (compound 7e).

$y alkyl substituents of imidazoles ,imidazoles Step A: A variety of the compound of the formula 2-chloro can be used to displace the alkyl moiety or 65 substituent of pyrazoles at elevated temperatures. IX producing compounds with the general structural formula [Xa] using quinoline as the 4-hydroxy core of the benzyl moiety. Step B: When removing the protective group of the core with the structural formula quinoline L: 2002; Hydrogenation methods derivatives are produced using standard hydrogenation methods

General Xe P3: Synthesis of the appropriate extended to form different D so that it contains the linking cleft P3 parts are synthesized

P3 and generally .olefin units — mega-ring metathesis are synthesized by double exchange

X terminals for double exchange by following the general schemes shown below (schematics) olefin contains (XT 5 XI) bonding moieties in Class A (does not contain carbon) This general synthesis is used to produce bonding moieties that are the basis of each

A(X heteroatoms) (scheme

1L chart X R* ANKE Xa I {= B A 7 AK >

Xb o 0 124 AA — nL Xc 0 0 R* AeA = 60 5 Xd 9 174 lap NHBoc Xe =m 1 to #0 alkyl J H = R* YYVY ty et al. in Evans lid. The synthesis was carried out according to the procedure described above. 5 f 7-011 vol 7 snort: “J.

Am.

Chem.

Soc. Or prepared by the procedures mentioned in Glad (Xa) primary carboxylic acids, and known and familiar pamphlets are available for those skilled in the technique.

Step A: Xa carboxylic acid is activated with pivaloyl chloride and then reacts with the anion of Evan's chiral auxiliary 4(S)-4-(phenylmethyl)-2-oxazolidinone as follows: Known Chemistry Tas (see reference cited by D.J. Ager 0.1.7 et al. in adcta 174700002 vol. Fe p.: AVY 99M, and the references cited therein) to obtain compounds with the general structural formula Xb

0 Step b: The process of adding azid at the alpha site is stereoselective as well using ctrizylazide for a chiral imide enolate such as those formed from compounds with the general structural formula Xb in the presence of a base <KHMDS Jie Tas is well known in technology (see reference above on D.J. Ager, D.

J.

Ager and his associates in the journal cdldrichimica Acta Vol. Fo p. NT 1949 AD and the references mentioned therein).

t-butyl formed once protective amine <SnCl was produced, step C: followed by p22:0-reduction; Catalyst B - 0 As well as these reactions are known Xe Intermediate compounds with general structural formula Al carbamate well into the technique. Mixture of ,11,0 with Jae Step D: Finally hydrolyze the non-compact catalyst under conditions Basic, in the general structural formula, amino acids, to produce bonding moieties of the type 3, amino acids.

‎XeY-

Alternatively, P3 moieties with the same general structural formula Xe are synthesized as follows

The procedure described in M. j. Burke J.

Burk.14 and collaborators in Journal

Am.

Chem.

Soc for vol. 071 p.: ¥=10V 717 Um shown in the XT chart. These compounds vary in number (Jam Cua) along the (-CHy) methylene bonding moiety. and does not contain a hetero-atom. JR at alkyl Sle gana to m) and in substitution 1 ranges from

XI chart COOE! COOE!

Jd roo NHAc for whom NHAC

Xia XIb

NHAc Yabah Mer © Mb ad xn ZF # coor:

H

Xie 2018

NHBoc NHAC rR 1 A ! Dr. i ho

AE coon A coor

Xe Xie alkyl H=R*,0 to 1 a number ranging from o =m of XIb in the form monoacid compound Step A: The monoacid compound is prepared as a basic cag pla by hydrolysis: © standard In Glad available diethyl 2-acetamidomalonate in the general structural formula aldehyde between Knoevenagel Step B: Condensation of the Novinagel type leads to the formation of an acetic anhydride 5 “pyridine such as acl having A XID A compound of formula Xe of formula 7010 has a stereochemistry of adjoining type 7 around the enamide double bond intermediate of the newly formed 01 as shown.

With Step C: enantioselective and locally selective regioselective catalytic hydrogenation of the intermediate compound of enamide of Formula 2020 to the intermediate of amino acid of Formula 6 using Burk's method. Step D: atom shielding nitrogen of the Xe acetamido derivative twice after adding the substitute t-butyl carbamate 0 before the hydrolysis of the acetate group in addition to cethyl ester under standard alkaline conditions to obtain moieties 3 with the general structural formula XIf synthesis Bonding moieties in class B The general structural formula for the bonding moieties in class B is NHBoc Rg & Ss f0-=X 01 H rice or CH, H = R, or CH, is used This general synthesis to produce bonding moieties containing oxygen or .sulfur

Scheme XII keen ANF C — A on spall L R P R R R NHBoc NHBoc NHR, Xie 20 m Ql X 0 0 7 ZN th ANF Apo — “afc aat ee K cg R ka R3 R! R R! NHBoc NHBoc NHBoc NHBoc 1 0 m0 08 S#0 =X rice H or CH; Boc-(L)- threonine methyl ester , Boc-(L)-serine methyl ester or Boc-¢(L)-allothreonine methyl ester Using allyl iodide in the presence of Ag,0 to produce a methyl ester with the formula XIIb Step B: The hydrolysis of methyl ester under standard basic conditions results in the bonding moieties of the ether type with the general structural formula XIlc (e(h). The step led prepares a sulfur isotope of the same 1-amino acid precursor of formula 118 (appropriately buffered as previously mentioned) and converts its hydroxyl ic gana to a group A

Easy removal of good Teaving group Jia) intermediate of XII tosylate using standard methodology known Tus in technique Step D: Substitution of tosyl moiety with GaY thioacetate anion leads to formation of thioester intermediate 1011 by inverting the inversion of the chiral central atom at the B-carbon atom. Step E: The hydrolysis of the thioester crack under moderately basic conditions results in a talc thiol with the formula XII. Step F: ASH is conducted. The thiol is readily cleaved under basic conditions using allyl iodide step g. Finally, (X=S) 31016 sulfide is obtained after methyl ester hydrolysis using standard procedures. Examples of Synthesis of Parts Cas 18 NHR is represented in IPL No. 0.4867.. Examples The present invention is illustrated in greater detail by reference to the following unspecified examples. Other proprietary methods of synthesis or dissolution may be found in IPL No. 00 /098467, Application 1 International Patent No. 048068/00 and the two joint pending applications with serial number RA 4/9 and 1 A/TTAATT, each of which is incorporated into this statement for reference. The temperature is shown in degrees Celcius, 688C06. The percentages of solutions express a weight-to-volume relationship, and the percentages of solutions express a volume-to-volume relationship, unless otherwise indicated. Nuclear magnetic resonance (NMR) spectra 9 were recorded on a Broker spectrometer with a frequency of 500 megaherts (MHz). The displacements were recorded as Abas (delta) (185: chemical). in part per million and referred to the internal deuterated solvent unless otherwise indicated NMR spectra were recorded for all final compounds (inhibitors) with a value of 0:(,50) (DMSO-dg) No

¢A for its salts of (tetrahydrofuran) TFA unless otherwise indicated. Flash column chromatography was performed on (SiO) silica gel according to Still's flash chromatography technique. oJ Org. Chem. and collaborators in W. C. Still tert-[Me;COC(0)]:Boc ¢benzyl:Bn Abbreviations used in the examples include °¢benzyloxycarbonyl:Cbz ¢bovine serum albumin (JY):BSA tbutyloxycarbonyl 1, 8- DBU: ¢3-[(3-cholamidopropyl)-dimethylammonio]-l-propanesulfonate :CHAPS: DEAD ¢methylene chloride :CH,Cl,=DCM 'tdiazabicyclo[5.4.0Jundec-7-ene ¢diisopropylethylamine :DIPEA 'diisopropylazodicarboxylate DIAD: ¢diethylazodicarboxylate 1,2- DME: 1,3-dicyclohexylcarbodiimide DCC: DMAP 1 ¢dimethylaminopyridine

DTT ¢ dimethylsulfoxide: DMSO ¢ dimethylformamide: DMF ¢ dimethyoxyethane ¢ diphenylphosphoryl azide: DPPA tthreo-l,4-dimercapto-2,3-butanediol or dithiothreitol tethyl acetate: EtOAc tethanol: EtOH ¢ethyl: Et ¢ethylenediaminetetraacetic acid: EDTA telectrospray mass spectrometry ESMS: ¢diethyl ether: Et,0: HPLC ¢O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate: HATU Vo mass spectrometry MS: thigh performance liquid chromatography, high-performance liquid chromatography

Matrix Assisted Canon-assisted laser ionization-time-of-flight MALDI-TOF ¢spcromety

Fast Atom Fast Atom Bombardment 3 (3 h): FAB ¢Laser Disorption Ionization Time of Flight (2-MES: ¢methanol:MeOH ¢methyl:Me ¢lithium aluminum hydride:LAH ¢Bombardment

N-:NMM ¢sodium bis(trimethylsilyl)amide:NaHMDS ¢[N-morpholino]ethane-sulfonic acid) Y-

Pr ¢ {N-methylpyrrolidine NMP: NMMO ¢ N-methylmorpholine oxide NMMO ¢ methylmorpholine tetra-TBAF: ¢ p-nitroanilide or 4-nitrophenylamino PNA: Succ ¢ 3-carboxypropanoyl: Succ ¢ propyl ¢ potassium tert-butoxide: tBuOK ¢tert -butyl-methyl ether: TBME ¢n-butylammonium fluoride

tris (2- TCEP : 2-(1H-benzotriazole-l-yl)-1,1,3,3-tetramethyluronium tetrafluoroborate :TBTU 'TIS ttetrahydrofuran :THF ¢trifluoroacetic acid : TFA ¢carboxyethyl) phosphine hydrochloride (TLC 'triisopropylsilane' TMSE 'thin layer chromatography' aminomethane hydrochloride :Tris/HCI ¢trimethylsilylethyl (GASO)15, 'Ry' denotes the °-rate of flow Pla ola i Example 1 Synthesis of 1 ) t-butyl-(1R,2R)/(1S,2S)-1-amino-2- homoallylcyclopropyl Carboxylate and 3 Br. 0 9 Nd ~ ~ ~ 0 9 f for the series symsis 0 rs 700 7 i A ag 0 0 IAL gd =~- A A ed ah SQ wp wf A ; A ' A ' or * 1 and ¢ or ye

a. 8.01 g (YY ET) of 1,2-dibromo-5-hexene was added and 87,; 77.71 g (mmol) of Hen-Dolha-N 1a successively reduced to a suspension of 65.08 g (77.7 mmol) of benzyltriethylammonium chloride in 0 © ml of 148011 aqueous (LL 3) , 0 was mixed vigorously at room temperature for 11 h, then diluted with FiO4, extracted three of je using 0 ml of CH,Cl, and the organic layer was washed. In addition, twice using 0 ml of water and twice using Or ml of a mixture of brine water in a ratio of 1/7, dried over 14850, and evaporated. silica using EtOAc with a concentration ranging from ? to 78 in hexane as the eluent, resulting in 7.44 g of the compound 1g (production rate of 0) FA (bd, 127.9 Hz, 2H), 1.25-1.33 (m, 1H), 1.46 (s, 9H), 1.19 s' H NMR (00013, 400 MHz): (s, OH), 1.47-1.60 (m, 1H), 1.75-1.82 (m, 1H), 2.14-2.22 (m, 2H), 4.93-5.50 (m, 2H), 4.96 1.48 (dm, J=10.2 Hz, 1H), 5.18 (dm, J=17.2 Hz , 1H). 0.75 (mmol) of potassium t-butoxide in 051 mL of anhydrous diethyl ether at 0°C and the reaction mixture was stirred at 0°C for 10 minutes. An ether solution of 1.48 g 017*(mmol) of compound 1c in 10 mL of diethyl ether was added and the mixture was stirred at room temperature for ¾ hours. The mixture was diluted with ice-cold water and extracted three times with 00010 mL of diethyl ether and the aqueous layer was acidified to a pH of 0.7* to ; using ice-cooled aqueous acid 016 of concentration 701 and the mixture re-extracted three times using 01 ml of 206 each time. The layer of 20/86 was washed twice using 100 ml of water, 100 ml of water A.

01 on the basis of (TAG) brine, dried over 14850 . and evaporated, producing compound 1d (production rate is the amount of raw material extracted.) 'H NMR (CDCls, 400 MHz): 5 1.51 (s, 9H), 1.64 -1.68 (m, 1H), 1.68-1.75 (m, 1H), 1.77- 1.88 (m, 1H), 1.96-2.01 (m, 1H), 2.03-2.22 (m, 3H), 5.01 (dm, J= 6.4 Hz, 1H), 5.03 (dm, J= 14.9

Hz, 1H), 5.72-5.83 (m, 1H). °

ES(+)MS: m/z 241 (M+H). 1.14 g to a solution of EEN (mmol) of 7 A) c. 800 μl 1, mL was added 1 anhydrous; then benzene mL of YO (74: ; mmol) of acid was added in 1 d and the mixture was heated to reflux temperature for diphenylphosphoryl azide (mmol) of 9,71) trimethylsilylethanol from (J se mM 9.6A) for three and a half hours. 7 mL of 0 was added later, and the stirring continued at the reflux temperature for additional hours. Then the mixture was cooled to diethyl ether at 10 mL from 10 °C. room temperature and evaporated to half of its original volume, diluted with 90.78 mL of brine water, dried (SlNaHCO; 71 mL of) and washed twice with V701 of EtOAc using silica over 14850, and evaporated. The remaining oil was absorbed on a gel as a translucent, yielding 4 g of pure compound 1H (production rate 0-hexane (AN) with a 'H NMR (CDCls, 400 MHz) of 0.03 (s) , 9H), 0.91-0.99 (m, 2H), 1.18-1.29 (m, 2H), 1.45 (bs, 11H), 1.56-1.72 (m, 2H), 2.02-2.18 (m, 2H), 4.12 (t , at 8.3 Hz, 2H), 4.93 (dm, J= 10.2 Hz, 1H), 4.98 (dm, J= 17.2 Hz, 1H), 5.07 (bs, 1H), 5.71-5.83 (m, 1H). To a solution of molecular THF in 1 concentration of t-BuNF (mmol) of 1.7) ml LY d. was added and the mixture was stirred (THF in ml of 1 in E1 cyclopropyl g (*7, mmol) of the derivative 4 hr then heated to reflux temperature for 10 min. 06 at room temperature for Yq os) divorce amine — and evaporated solvent carefully under reduced pressure (due to high volatility oy extra caution during solvent evaporation).The crude ore was re-dissolved in 100 mL of BIOAC and washed twice with 506 mL of water, 506 mL of brine , dried over MgSO, and carefully evaporated the solvent again. Crude product 1f (as a mixture of the enantiomers or 's' and pedisen 1') was used to couple the P2 proline derivatives without further purification. Separation was carried out for fraction 0 2 having the desired stereochemistry at 71 readily at this point using flash chromatography (Example 1 Part (SY)

P2 moieties Example: “c) ) Boc-4(R)-[(7-methoxy-4-quinolinyl) oxy|proline synthesis N. CO 0 , om wl omy — — = iv

Ta 9

Bod CO, CH, —aY > \ 0 According to the method described as reported by M. (a) 4-Hydroxy-7-methoxyquinoline yas as reported by Y. s. Choi Olmstead, K. 16. PK. jk. Olmsted Chun, 14. W. W. Sean A ov

Y. 01,5©; What ela about c. Oh. Lee .0 flee, C. Ma ela on C-K. Lee, C.-K. h. Kim.

Bioorg.

Med.

Chem.

Lett vol Vv p: VAR 9911 M. A solution of 1.88 g V4 VY (mmol) of compound “b and 7.4 mL (71.47 mmol) of DEAD was added in anhydrous THF to a stirred solution of 7.17 g (0° VY)mUsa of buffered iY cis-hydroxyproline and 5.17 g (17 mmol) of triphenylphosphine in 001 ml of anhydrous THF at 0 °C in nitrogen atmosphere and the reaction mixture was left to warm to room temperature and stirred for VE 1 hour.Then the THF was evaporated and the pure product al was isolated after flash column chromatography using 1 His concentration is 7.0

In EtOAC as the solvent yielded 1.0 g (yield 7705).

'H NMR (J0W) 400 MHz): 1.44 (s, 9H), 1.65 (bs, 1H), 2.34-2.43 (m, 1H), 2.63-2.76 1 (m, 1H), 3.78 (s, 3H), 3.75-3.85 & 3.89-3.99 (2m, 1H, rotamers), 3.95 (s, 3H), 4.51 & 4.60 (2t, J= 8 Hz, 1H, rotamers), 5.15 (bs, 1H) , 6.53-6.59 (m, 1H), 7.12-7.18 (dd, J=8.9 & 2.2 Hz, 1H), 7.36 (d, J=2.6 Hz, 1H), 8.03 (bd, J= 9.2 Hz, 1H ), 8.65 (bd, J= 5.1 Hz, 1H). Example? ("c) 2-ethoxy-4-hydroxy-7-methoxy quinoline Synthesis Veo ! - A # COOMe © 1 N 8 with - A B A ot

Katz on Katz sla lad as described i¥ methyl-p-methoxyantranylate Synthesis was performed by Ahir 167 lm. FAL p: VA Vol. Org. Chem and collaborators in the journal On Bakkar ela a modification of a procedure described in—aY quinoline is the general synthesis of a derivative M. 1: 770, 1: Part B p.: cIndian Journal of Chemistry and collaborators in the journal Baccar ml £,Y !a in methyl-p-methoxyantranylate a. Dissolve 4 g (7 mmol) of 0 mmol) from a 0,000 μl solution, then add 0 mmol striethylorthoacetate (from Yo,¢) and heat the resulting mixture at diovane reflux temperature its anhydrous concentration; Standard in HOI in a vacuum, yielding 4.57 g of the product “B in A volatiles” h. Then the volatile substances were evaporated 19 Bad. Photo of a reddish-yellow oil. Use as is in the next step (production rate). B. 0 to a solution of molecular THF in 1 ml of LIHMDS concentration (3, eq.) of YY b. THF was added at a temperature of 1 ml of VE in OF substrate Approximately 7 mmol of substrate and the low-temperature bath were removed shortly after nitrogen atmosphere (sp YA) addition, and the mixture was left to stir at ambient temperature for 1 hour; after this time ml From 11120005. The resulting mixture was stirred until the substance VV disappeared. Another amount of direct co, Ry 000 was added by the initial TLC ratio (EtOAc) completely (after one hour) by and the mixture was concentrated in dioxane of 1101 ml. Its concentration is in 01 titer L (Y) was added to the product - R, aqueous NaH,PO, ml of 10 ml of 21086 and 10 vacuum. The resulting paste was grinded from a mixture of two molecules and exposed For sound waves, an abundant precipitate was formed, collected by filtration, and 1 gram concentration of the desired product was washed *g in the form of a beige solid (7.111 rate with water and dried vintage ye (HPLC) by purity > 7.94 Production reached 784 for the two steps » Ratio 'H NMR (400 MHz, DMSO-dg) (ppm): 7.88 (d, at 8.9 Hz, 1H), 6.98 digits 1H), 6.89 (br.d , J= 8.6 Hz, 1H), 5.94 (br.s, 1H), 4.30 (br.s, 2H), 3.84 (s, 3H), 1.34 (t, J= 7.0 Hz, 3H). Except

Example + Synthesis of (ot ) 4-hydroxy-7-methoxy-2(3-methyl-1,2,4-0xadiazol-5-yl) Quinoline N COOMe i 0 N, COOMe 0 Gp 2 OH OMEM it wt - 7 | P N Sx 0 N, 0 Ny N c. N ZZ OH OMEM af —¢ Li 0 mg (94 mmol) of NaH (dispersed at 758 ppm in mineral oil °) was added to a solution of 1 g (17 mmol) of 2-carbomethoxy-4-hydroxy-7-methoxyquinoline a (The method of its preparation is described in my patent applications No. qotyY/m” and No. 004 /0.) at 01 ml of DMF in nitrogen atmosphere and stirred the resulting mixture at ambient temperature for one hour; Then 00 μl (0,448 mmol) of MEM chloride was added dropwise and the resulting mixture was stirred at ambient gall temperature for an additional 19.0 ye h. The reaction mixture was diluted with 100 mL of FtOAC and washed with 50 mL of (HO).

of brine water; It was dried over (MgSO), and concentrated in a vacuum resulting in 1.77% of the raw material isolated from the reaction mixture. B. ¥ ml of THF was added to a mixture of 0,000 mg of freshly activated molecular sieve eyes diameter °€ freshly activated 4A molecular sieve and YEA mg (7.75 mmol) of .acetamidoxime and the resulting mixture was stirred for 0 V0 min in a nitrogen atmosphere at ambient temperature, then YE (mg YE, ¥ mmol) of NaH (dispersed 77680 in mineral oil) was added ) in batches. The resulting suspension was stirred at ambient temperature for one hour; Then, ©0001 mg (1.07 mmol) of ester was added to a solution in ©mL of THF and the resulting mixture was heated at reflux for 1 hour, then filtered over Celite and washed In ¥ batches of 86/20 by 71 ml each and concentrate in a vacuum. The resulting crude mixture was purified by flash column chromatography (using EtOAc (concentration)/000), yielding pale YOY of product af as a white solid (yield rate 775). c. 1 mL of aqueous HCL (concentration 1 M) was added to 171 mg 84F (+ mmol) of MEMether | 0 ;c in; ml of THF and the resulting mixture was stirred at ambient temperature for one hour and then diluted with 0 © ml of 11:11:70, aqueous (concentration 1 molecular). The resulting solid was filtered and pulverized with EtOAc, filtered and dried, producing a pile of 00 of the desired product (;d) in the form of a white solid (production rate was (VY MS (ES+) 258 (M+1), (ES) -) 256 (M-1).'H NMR (400 MHz, DMSO-ds) § (ppm): 8.03 (d,J= 9.2 Hz, 1H), 7.38 (d, J=2.2 Hz, 1H) , ¥.Hz, 1H), 6.85 (br.s, 1H), 3.88 (s, 3H), 2.64 (s, 3H). 8.6 p1 (d, 7.06 YYVY ov Example E D ) 4-hydroxy-7-methoxy-2(5-methyl-1,3,4-oxadiazol-2-yl) quinoline Synthesis 6 N COOMe 0. » _N CONHNH, —_—

OMEM OMEM ot I - - 0 } N, = A } N, N, OE J N = nN Nn” hi re SF

OR OMEM

MEM = R:—a0 wo

H=R:s0 to 70 mg (1.20 mmol) SLY hydrazine mmol) from V,A) µl OV a. Added from KB base material © ML by 02001©. The resulting solution was heated at the reflux temperature for a period (as a quantitative crude product) in the form of jo material, mg of product Vt, and then concentrated in the vacuity of the finished product, a yellow solid that was used as it is in the next step. At the degree of triethylorthoacetate in © ml of fo mmol presumably of the compound, 0.559 Pa. Heat 01 001 mL and then dilute the resulting mixture with nitrogen to 1101 C in an atmosphere of 0 0 S aqueous; 005 mL of brine and dried over NaHCO; of 510/88 and washed with 0 * ml of no oA

EtOAc was concentrated in vacuo and pure by flash column chromatography (by liquefaction with MgSO, mg of compound B in the form of yellow oil (production rate TOR yield 76000 concentration for each of the two steps). 771. MS (ES+) 392 (M+1), (ES-) 390 (M-1). mmol) of compound 0 (at 6600 °C in extreme vacuum for ACY) mg PVT in hot air 0

EtOAc by flash vertical chromatography (by chromatography using By hours and a half A (+, mg of a mixture of compound #B) (production rate was 076 B-1116 Vantage (7) er, its concentration was 1 ml of 01.3 b-VY mg of compound H (corrected production rate was WA mg (4 mmol) of compound 20 in 18 M) was added to a solution of 1 aqueous (its concentration HC in the vacuum of THF min.) and evaporate 0 and stir the resulting mixture until the reaction is complete (for THF; 1 ml of sonic product; Glad) molecular ales). 1 was added ?ml of 11211.00 aqueous (its concentration is 20 mg of the desired product in the form of VO), filtered, and the solid was dried in a very vacuum, yielding a beige color solid (IVY) (the production rate was

MS (ES+) 258 (M+1), (ES-) 256 (M-1). 'H NMR (400 MHz, DMSO-dq) e (ppm): 8.03 (d,J=9.2 Hz, 1H), 738 (d, ]=2.2 Hz, IH), | 0 7.06 (d, J= 8.6 Hz, 1H), 6.85 (br.s, 1H), 3.88 (s, 3H), 2.64 (s, 3H).

Example > Synthesis of 4-benzyloxy-2-(chloro)-7-methoxyquinoline (1E) HCl NH, Da 0 NH, 0 NH; N 0 NC 0B OH kb i N, a } N oH 1 N, NH; > 8 — = OBn OBn OBn C Ea AH A. Chill YO g (oY mol) of commercially available meta-anisidine in Av ml of dioxane to 0 °C and add VO ml (7©; mol) of HCI Anhydrous (its concentration; standard in dioxane), then 00 ml of BLO was added, and the sad stirring continued for one hour. Then the beige-colored solid was filtered and dried in a vacuum, resulting in 1.88 g of salt “A” (the production rate was (AA b. To this salt was added 71.1 ml (oY mol) of ethylcyanoacetate and the mixture was heated in a flask fitted with a distillation head and a collecting flask to a temperature of 1 heat from NYA 00 m and collect the resulting ethanol to monitor the occurrence of the reaction * and when collecting 9 mL of ethanol (theoretical amount (Jo 1) Y' stop the heating and cool the reaction mixture to room temperature; Diluted with a mixture of 0 ml of water and Yoo ml of EtOAc then stirred and added Yeo ml of 0 aqueous NaH,PO and after further stirring for 1 hour then filtering and drying yielded 1 g of compound 1b (purity 0.£AE) as a yellow solid

(production rate was about 100) used as is in the subsequent reaction. c. 00 g (OV,A) mmol) of compound “B” was added in 100 ml of DMF at zero degrees Celsius to 7,728 g (110.71 mmol) of NaH (70% dispersed in mineral oil). Then the ice bath was removed and the mixture was stirred at ambient temperature sad for one hour, then V1 ml (1 mmol) of benzyl bromide was added and the reaction mixture was stirred for 11 hours. Then the solution was diluted with a mixture of 701 mL of EtOAc and 1077 mL of hexane and the solid formed was filtered off; and triturated with 1,000 mL of NaHCO, aqueous saturated; and washed using water; and 1010 mL of a mixture of hexane and 2026 in a ratio of 1:1 and dried in a vigorous vacuum. J produced © g of product 1g in this manner (purity 751) as a yellow solid 0 (yield rate 770) VA ml YA (mmol) was added from iso -amyl nitrite to 1.17 g (4.0 mmol) of compound 7g in 71 ml of acetic acid and the resulting mixture was stirred at ambient temperature and controlled by HPLC and an additional 1.7 ml was added 1.07 mmol) of iso-amyl nitrite after 2 hours and the mixture was allowed to oscillate during 0 hours (production rate was 7481 of product and 77 of substrate as determined by HPLC and 100 ml of water was added to the suspension Vo The product, which Lad was filtered after, and the brown solid was dried after collecting it in a very vacuum, resulting in 8 g of product 1D (with a purity rate of 757, the production rate was (VY d. 1” ml VEY was added) mmol) of phosphorous oxychloride to #1 g (¥9,£ mmol) of compound 1D and the resulting mixture was heated at the reflux temperature for one hour, then diluted with Je Or 151086 and quenched at zero degrees Slowly using +10 ml of NaOH Sle 0 (concentration 1 N) to a pH of 4. The two layers were separated, and the organic layer was dried over 14850, and concentrated in a vacuum, resulting in a brown solid that was purified by flash chromatography (by chromatography). A mixture of hexane s EtOAc with a concentration of 11/.4 mg of product 7e (purity >99/) was produced as a yellow solid (production rate of 777).

TN

'H NMR (400 MHz, CDCL): 8.07 (d,J= 9.2 Hz, 1H), 7.5-7.4 (m, 5H), 7.29 (d, 1 2.5

Hz, 1H), 7.12 (dd, J=9.2, 2.5 Hz, 1H), 6.73 (s, 1H), 5.26 (s, 2H), 3.92 (s, 3H). 1 Example 4-hydroxy-2-(4-methyl-1- ¢ lab ) 4-hydroxy-2-(1-imidazolyl)-7-methoxyquinoline Synthesis of 4-hydroxy-7-methoxy- 2-(1-pyrazolyl)quinoline ¢ (av ) imidazolyl)-7-methoxyquinoline ° AH ) 4-hydroxy-2-(3-methyl-1-pyrazolyl)-7-methoxyquinoline LAO); in ) = lS x joo]

Ba=R:ivV 8 OR H=R:qv { 1,

N i 8 . : e o. N, a b 6 N, NS Ajn whip

OBn 8 OR H=R:v uh ~N H ASS { > a \ — 7 a" 1

Bon =R:-avY ru( OR H=R:y 6 | Neo NN

Ba=R:jV or H=R:V a

+27

a Heat EYY mg (41 mmol) of compound 7E ony mg (5.88 mmol) imidazole at 10 °C for 1 h. Then the mixture was diluted with M208 and washed with water and brine; It was dried over 0.04880 and concentrated under reduced pressure to produce pile YY from compound A! (with a purity of 747) in the form of a yellow solid (production rate of £80 (914 mg + .87Y (0 mmol) was cleaned from Compound IV using 4 pide of palladium (borne on 70% carbon) in a mixture of ©ml of ethanol and ©ml of THF and placed in an atmosphere of hydrogen at a pressure of 1 atmospheric pressure, and after 1 and a half hours of stirring at ambient temperature, the reaction mixture was filtered and washed with a mixture of chloroform and p0, and 01 mg of QV was concentrated.

(purity 4997.7/) as a yellow solid (production rate 707) MS (ES+) 242 (M+1), (ES-) 240 (M-1). ye

'H NMR (400 MHz, DMSO-de): 8.51 (s, 1H), 8.03 (d,J= 8.9 Hz, 1H), 7.93 (s, 1H), 7.23 (d, J=1.9 Hz, 1H), 7.15 (s, 1H), 7.12 (dd, J= 9.2, 2.2 Hz, 1H), 6.92 (br.s, 1H), 3.91 (5, 3H). Preheat YO mg ATV (mmol) of compound 7e f ;; mg (£18 mL) of 4-methylimidazole at 10 C for yo h. Then the mixture was diluted with EtOAc, washed with ve water and brine; dried over MgSO, and concentrated under reduced pressure to produce Crude product containing a mixture of S-methylimidazolyl s 4-methyl isomer in a ratio of 1:01 in the order of 0. VAT separated mg of the main putative desired isomer 11g (purity 99/) which is White solid (rate of yield 87/) of 11 mg of a second more polar fraction containing a mixture of zyminer and 5-methylimidazolyl -4 Y. (rate of yield Report (YY) by chromatography (by blotting with EtOAc concentration ++ (1) and clean 117 mg EVY (+, mmol) of compound/Ag using FF mg palladium (portable on carbon by 70) in a mixture of 7.4 ml of ethanol and © ml of THF and placed in an atmosphere of hydrogen at a pressure of 1 atmospheric pressure. After 18 hours of stirring at a ay concentrate “methanol g chloroform” with a mixture of Ju 5 “Jeli filter haaall gall mixture at temperature of compound E (with a purity of 749) in the form of a white solid, pile 118 produced ( AA (production rate as 'H NMR (400 MHz, 1480-2: 5 8.42 (brs, 1H), 8.01 (d.J= 9.2 Hz, 1H), 7.64 (brs, 1H), 7.21 (brs, 1H), 7.10 (d,J= 8.9 Hz, 1H), 6.89 (brs, 1H), 3.90 (s, 3H), 2.20 (5, 3H). . mg (+118 mmol) of compound 7e and 0560 mg (©,07 mmol) of VAE c. Preheat aqueous NaOH and wash with EtOAc for 1 hour. Then the mixture was diluted by 11 m for a period of 11 0 at pyrazole and concentrated under low pressure to produce a crude product MSOs brine; It was dried over a standard slay (1) concentrated in EtOAc hexane by chromatography using a mixture of (i) of compound 1e in the form of a pale yellow solid (production rate pile 017 vintage) (V:¥ Ve mg of 01 mmol) of compound LAH using 711 (017 mg) and clean (70+ ab

THF ml of Ys ethanol (provided to 0 by 5) in a mixture of ¥ ml palladium atmospheric pressure. And after © hours and a half of stirring at 1 hydrogen pressure and placed in an atmosphere of methanol § chloroform concentration the temperature of the surrounding atmosphere; filter the reaction mixture; It was washed with a mixture of 1 mg of the compound LAO (with a purity of 799) in the form of a yellow solid VV, with a yield of 0 (728) (the production rate was

MS (ES+) 242 (M+1), (ES-) 240 (M-1).

H NMR (400 MHz, DMSO-dq): 5 8.72 (d,J= 2.5 Hz, 1H), 8.31 (s,1H), 8.00 (d, J=8.9 Hz, 1H), 7.83 (br.s, 1H ), 7.43 (brs, 1H), 7.24 (br.s, 1H), 7.10 (d, J= 8.6 Hz, 1H), 6.59 (br.s, 1H), 3.90 (s, 3H). 7 mmol) of (pale, 74 mmol) of compound 7H and 44 VTE) pide YIV d. Preheat an hour. Then the mixture containing the compound YY was diluted at 1011°C for 1-7 ml of 4-methylpyrazole using (10 lys) in proportion to the benzylated product H and the benzyl product de-1

> 790 nm (with a purity of benzyl) from the pure product, de-axle 111 of 10/86 and filtered by Vintage. VA ¢ as a white solid (production rate was H NMR (400 MHz, DMSO) -dg): 8.58 (d,J= 2.6 Hz, 1H), 7.98 (d, J= 9.2 Hz, 1H), 7.25 (br.s,1H), 7.20 (s, 1H), 7.04 (br .d, J=9.2 Hz, 1H), 6.38 (s, 1H), 3.89 (s, 3H), 2.30 (s, 3H).

A Example 5 (sh) 4-hydroxy-7-methoxy-2[4 (2-isopropylaminothiazolyl)] Quinoline Synthesis Using the Same Scheme as 2-alkylaminothiazolyl Note: [Different substituents of others prepared]. ‎ alkyl thioureas — replacing Glad

NH; NH; 7 lakh | O.C. 0

A

PN

A i w x “Hr + b 5 3 I to — ~~ 0 0 mg h aA no

1

NH DY

M Hb + J. | ~~

HO, J Pe NH, 8 ©0 y nl 0 0 6 AH M Hd 1 d 8 =< ~~ or

N

0 is to compound A identical to that described in the meanisidine leaflet The method used to convert A

AYT=ALY p; FY vol. of Med Chem. and his collaborators in the journal 7.3. Brown F. j. Brown to avoid chromatography. A fluidized phase of Jae QM 5 + purification procedure was treated with coal silica 8021 and gel (MgSO, containing the desired product using a mixture of 0 ether — expected mass basis). After filtering on Silite; Grinding the product (Gs 8 Je) with a purity of more than 7949 (as proved by the aly brown color Ada) and compound “A” was produced in the form of a substance (HPLC B) Hb and © gm of isopropyl thiourea mmol ( of 0 ) b. Heat a suspension of ¥00 g (molecular concentration 1) of dioxane mL of Yoo eq) in 1 (H) 3-bromopyruvic acid Ve a

As m. When the reaction reached 86 °C, the solution became (GW) after which the product rapidly precipitated in the form of a white solid. After two hours of heating, the solution was cooled to room temperature and the white precipitate was filtered, resulting in 7.01 g of compound “D” with a high purity of more than 798 as proven by NMR (the production rate was 194).

o c. Cool a mixture of £,A0 g (YAY mmol) of carboxylic acid .hd and g of its derivative (1 fA aniline equivalent) in YO ml of pyridine (moM 7) to -. Ny (and during the canal the clear solution became partly Gla). Then 3.86 mL of (phosphorous oxychloride equivalent) (Y,1) was added slowly over a period of ∼ minutes. The reaction mixture was stirred at -1 C for 1 hour; The bath was removed and the Bay reaction mixture was brought to temperature

> The room. After an hour and a half the reaction mixture was poured into Bll and the pH was adjusted to 11 using aqueous NaOH (concentration ¥ ge), extracted with CHCl, dried over MgSO, anhydrous; filtered and concentrated in vacuum. Then the beige solid was purified by flash chromatography (by chromatography using BtOAc in hexane at a concentration of 45/) and 1.17 g of compound 4e was produced in the form of a pale yellow solid (production rate of AVY

Dr. Heat a solution of 7.47 g (YV,T) mmol) of BBUOK in 40 mL of anhydrous {BUOH (M4; distilled from Mg) to reflux temperature. Add 8 g (0.8 mmol) of compound 4e in batches over 0 minutes and stirred the dark red solution formed at the reflux temperature for an additional 1 minute (and the completion of the reaction was monitored by “HPLC”). The mixture was cooled to room temperature and HCI was added. (concentration; standard in

1,9 dioxane ¥- equivalent). Then the mixture was concentrated in a vacuum to ensure that all 1101 dioxane s were removed, and the product was re-dissolved twice in CHCl and dried in a vacuum to finally obtain 1,167 g of HCI salt for the compound “and in the form of a beige solid (with a purity of 797 as specified by HPLC and then pour the product into a pH buffer — NaH,PO,) phosphate of concentration 1N

No

Tv for sound waves. The solid, beige Uae (with a pH of about m), was filtered and dried in a vacuum, yielding 1.74 g of the compound (with a purity of 7951 as specified by B. 7A) in the form of a beige solid (production rate). (HPLC 'H NMR (400 MHz, DMSO-d): 8 8.58 (d,J= 2.6 Hz, 1H), 7.98 (d, J= 9.2 Hz, 1H), 7.25 (br.s,1H), 7.20 (s, 1H), 7.04 (br.d, J=9.2 Hz, 1H), 6.38 (s, 1H), 3.89 (s, 3H), 2.30 (s, 3H). ) 4-hydroxy-7-methoxy-2[2 (4-isopropylthiazoyl)] quinoline was synthesized using the same synthesis scheme as 2-(4-alkyl)-thiazolyl Note: Different substitutions were prepared from one another. a-bromoketones — replace the compound with 0 9 following << mab. splly = / 6 5 14 ap —4 0 0 7 ~ 0 NH . 2 0 "" a 5 So NA a4 o.0 ah iA oH

LOL

Ss or 01 no

a. added 79,; ml AY (mmol) of 1 (Br equivalent) drop by drop to a solution of + g (¥ 4 mmol) of 3-methyl-butan-2-one in 100 ml of methanol when -. m over a period of £0 minutes. Then the resulting mixture was stirred at room temperature for 90 minutes. Pentane was added and the solution was washed with 5N aqueous NaHCO, then the organic layer © was dried over 1.90 SLY, filtered and concentrated in a vacuum. The resulting crude yellow oil was used; The compound “ot” was obtained without further purification. A solution of VA was stirred at 0.03 g (mmol) of VV,0 5 fa ethyl thiooxamate mmol of the bromoketone derivative at 0 L for 1 hour. Then the mixture was concentrated in vacuo and later purified by vertical flash chromatography using a mixture of FIOAC in hexane 710 as the oleophobic material; Yield 1460 mg of product 4g (production rate 01 was (7X A) b. A solution of pile 001 (10 mmol) of compound 4g was treated in VT ml of a mixture of THF/MeOH/ H,0 in a ratio of (0:0:7) using 4 mg (0,¥ mmol) of (1 LIOHH0 equivalent) at room temperature for © hours. Then the pH was adjusted to a value of 1 using 11 M HCL and the mixture was concentrated to dryness in a vacuum to obtain 4 0 acid acid; which was used directly in the next step without further purification. = cold solution of 890 mg (° diene mmol) of 4-methoxy-2-amine-acetophenone (intermediate compound 8a) and 0960 mg (0 Vt mmol) of the 1d1 carboxylic acid derivative (eq.) in 01 mL of pyridine to -10 M. Then add Ye mL TVA (mmol) of POC, (1.1 eq) point by point during an Agia period of © minutes. The resulting solution was stirred in a dreamer for two hours. The reaction mixture was quenched by adding 11.0 and the mixture was concentrated in a vacuum. The ore was poured into a saturated aqueous solution of NaHCO; EtOAc was extracted and the organic layer was dried over MgSO, anhydrous; filtered and concentrated in vacuo. The crude product was purified by flash column chromatography using a mixture of EtOAc in hexane concentration 7 As a liquefied material, 7560 pales of compound no

(9=—) as a white solid (production rate was 51/). d. OVA of 8:01 mg (7.1 equivalent) was added to a suspension of 706 mg VY) mM (Use of compound 4e in 11 mL of anhydrous 080011. The resulting mixture was heated to VO 4 for seven and a half hours. Cool the solution to room temperature py by adding 0.¥ mL of HCI (concentration 0 0 Nitrate in (dioxane) and concentrate the mixture in a vacuum and pour the resulting ore into a solution of NaH,PO, (its concentration is 1 Mn) and filter. Then the solid was grinded with a small amount of 250/8, filtered and dried in a vacuum, resulting in 770 pale of compound 1 and as a beige solid cals (production rate was .)/461 (bs, 1H), 7.6 (br.s, 1H), 7.51 (br.s,1H), 7.43 (brs, 8.00 h NMR (400 MHz, DMSO-dq) 1H), 7.29 (br.s, 1H), 7.14 (br.s, 1H), 6.95 (br.s, 1H ), 3.90 (s, 3H), 3.15 (m, 1H), 1.33 (d, J=0.4 Hz, 0-00 6H).

Ye 01 JU (99 0 ) 4-hydroxy-2(1-methyl-2-imidazolyl)-7-methoxyquinoline Clearance _o NH, 0 j iA 0 release A Co Yb Lba N “ Aless ~o

Be OH i \ 0 ive c0

Oh

L

_N d0 ml from 1117 to 001 in iY + N-methylimidazole mmol) of 71 (a cold solution of © g eq) 1 molecular point; Y,0 concentration BLO (solution in n-Buli d Helam. 754.4 mL of pour was added in batches over a VA= min. The resulting mixture was stirred for 0 min at Ye point through annealing. The mixture was stirred Heterogeneous for 2 hours and left until it reached CO, an excess amount of titer) until it reached pH ©" and then 1 room temperature. And 1101 (its concentration) was added to remove EtOAc. The aqueous layer was separated and dried. The resulting ore was extracted in this way.

except salts); Then it was dried over 50, filtered and concentrated under reduced pressure. 7.7 g of a white solid 101b (production rate was (TA b. 3 solution of 44? mg (71 mmol)) of 4-methoxy-2-amino acetophenone m and 00 mg of its derivative 1) “+ carboxylic acid eq.) in 01 ml of pyridine 10-00M. Then YEE μl of ,1 (POC equivalent) drop by drop was added over © minutes. The resulting solution was stirred at Vem for two and a half hours. Then water was added and the mixture was concentrated under reduced pressure. The ore was poured into a saturated solution of NaHCO, extracted with EtOAc, dried the organic phase over MgSO, filtered and concentrated under reduced pressure. The product was purified by chromatography using silica gel (chromatography using a mixture of EtOAC and hexane concentration (YO 0) from a pale yellow solid 10g (production rate of 781). c. added pale 71 of 80016 7.1 (equivalent) to a suspension of 9000 mg VA (mmol) of 0g base material in A mL of 080011 then heat the resulting mixture to 78°C for 1 hour . Leave the solution to reach room temperature overnight and add ©,? ml of HCI (concentration ¢ in 0 Mn) dioxane and the mixture was concentrated under reduced pressure and the resulting ore was diluted (EtOAc — Vo) and NaOH (concentration 1 Mn) was added until the pH reached VY and separation The organic phase was dried over MgSO, filtered, and concentrated under reduced pressure, yielding 0£ V mg of the 0d compound as a pale beige solid (production rate of 771' H NMR (400 MHz, -1480). 4: 57.99 (d, J= 8.9 Hz, 1H), 7.49 (s, 1H), 7.37 (s,1H), 7.18 (s.1H), 6.92 (d, J= 8.9 Hz, 1H), 6.31 (s, 1H), 3.87 (5, 3H), 3.84 (5, 3H).

In Example 11, the synthesis of 1) 4-hydroxy-2(1-pyrrolyl)-7-methoxy quinoline b “behb OH 0. — ZZ No N NH, ivy OH 0 2 0 hy i . A solution of 1 g (¥0,© mmol) of a substrate (11) obtained from 0 - 0g compound after hydrogenolysis of a benzyl group using a J sans palladium catalyst was heated at 0 H 79 percent in a mixture of ethanol (1117) and 14 mL of Y (1) 2,5-dimethoxytetrahydro furan eq.) in glacial acetic acid with regurgitation for ; An hour and a half and leave until it reaches room temperature. Then concentrate the mixture under reduced pressure. The ore was diluted — methanol and 1 aqueous was added with a concentration of 1 N until the pH reached ye. LV and the product was purified by chromatography using silica gel (by chromatography using a mixture of 1 al and CHCl, its concentration 7 The concentrate was re-adsorbed on a silica gel and 060 mg of compound 1AP was produced as a white solid (production rate was .7\Y). H NMR (400 MHz, DMSO-d) : 5 7.98 (4, J= 9.2 Hz, 1H), 7.64 (E) 2H), 7.18 (d, 1-5

Hz, 1H), 7.05 (bd, J= 7.9 Hz, 1H), 6.88 (bs, 1H), 6.32 (5, 2H), 3.90 (s, 3H). A vy 11 Example (D11) 4-hydroxy-7-methoxy-2-(6-methyl-2-pyridyl) quinoline Synthesis J 58 A

BN0

A

LA ~ — LA - - LA

N N N

0 0 0 ivy “ab ° \ N N

OH avy ml +,0Y +5 IVY 6-methylpicolinic acid mmol) of 0,0)a preheat 11; mg for two hours. benzene eq) with reflux in (ml) of (Y,£) SOCL (mmol) of 0,7) pentane. The reaction was carried out in a vacuum and the ore was pulverized by made, and the solvent was removed and an excess amount of 500 mmol) of 7.1 (pale ove) and the solid formed was filtered off and the filtrate was concentrated, producing a cap. acid chloride ml 6 3 iA aniline mmol) from YX, A) mg vie added a solution from old

CHCl, 10 mL in DMAP mg (50+ mmol) of DIPEA and 11 mmol (AYO) ve ab in © mL of © at gestational zero. And stir the crude Y acid chloride into a solution for an hour. The volatile components were removed in the vacuum of VT reaction mixture at room temperature for a period of 70 minutes, then with water and brine. then NaHCO; The solution was mixed twice with Ju 5 EtOAc and the concentrate was dissolved in the organic layer was dried above 14880, and concentrated in a vacuum. The mixture was purified by flash chromatography as a liquefied material. produced 4980 mg 7:1 columnar EtOAc/hexane ratio; Using a mixture of (7AY) from P201010 "Ag" (production rate reached

00 c. 0 mg (7.47 mmol) of 8:01 was added to a suspension of 96; mg VV (mmol) of amide 71g in 10 mL of 0011© and the mixture was stirred at L for 1 hour and then at room temperature for 1 hour. Then the mixture was poured into 175 mL of a pH buffer — phosphate (pH-V) and stirred for 10 minutes. The solid was triturated twice — ethyl acetate and the organic phase washed with brine; dried over

MgSO, ye and concentrate in a vacuum. The resulting solid was triturated with 206 nitrate, yielding 7717 mg of MY quinoline derivative (production rate was A 10 (. (s, 3H), 3.94 (5, 3H), 6.85-6.88 (2d, J= 8.68 & 9.5). 2.68 E' H NMR (CDCl, 400 MHz): Hz, 2H), 6.94 (dd, J= 8.9 & 2.2 Hz, 1H), 7.27 (dd, J=6.7 & 1.9 Hz, 1H), 7.73 -7.79 (m, 2H), 8.28 (d, J= 8.9 Hz, 1H), 10.3 (brs, 1H).

Yo

VY Ex. (9 \Y ) 4-hydroxy-7-methoxy-2-(5-methoxy-2-pyridyl) quinoline synthesis 0 0 > oO A

N ° 7 NT N 0 6 0 ivy “ab c 0 y c ©. } dl | SF oH

NY

a. £,Y mL of NaOH (concentration? molecular) was added to a solution of 677 mg (7.77 mM (Use 0.0) of NY in MeOH and the reaction mixture was stirred at room temperature b. The solution was aan aan using 0.1 ml of HCI (concentration 1 M) and concentrated to obtain the compound oo) which was used in the next step without purification. b. Ou was added 9.9 mg mmol) of mA aniline to a solution of about both mmol of the crude compound (RIT) in YO ml of Shi pyridine solution to -75 M before adding 0 1.79 ml VV) millimoles) from 2001. The reaction mixture was stirred at -10°C for one hour and then at 0°C for two hours. Then pour the mixture over the water and extract Ode to three vi 70 concentration NaHCO; The organic layers were washed after mixing them with EtOAc several times with brine water; It was dried over 0.14850 and concentrated in a vacuum. The raw material was purified by flash chromatography using amalgam of VY as the liquefied material, resulting in a 7:1 ratio of vertical EtOAc/hexane; Using a mixture of (700 (rate of production was 11 C) amide 0 G. 0 mg (0.11 mmol) of 0:8 was added to a suspension of 0 mg NY 0°) mmol ) of amide” 1g in 10 mL of anhydrous 1120011 and stirred the mixture at VO for 1 h then at room temperature for 110 hr. The reaction mixture was poured into 1 VO mL of buffer For pH — phosphate (pH was ¥) and stirred for yo min. The solid formed was filtered and triturated with BIOAC, producing pila YOu from EY derivative (production rate was NY quinoline | 0 "H NMR (DMSO, 400 MHz): , 3.86 Hz 3H), 3.94 (s, 3H), 6.72 (bs, 1H), 6.91 (dd, I= 8.9 & 1.9 Hz, 1H), 7.54 (d, I = 1.9 Hz, 1H), 7.60 (dd, J=8.9 & 2.9 Hz, 1H), 7.97 (d, J= 8.9 Hz, 1H), 8.21 (d, all 8.6 Hz, 1H), 8.48 (d , J= 1.9 Hz, 1H).1 4 Example (= ¢ ) 4-hydroxy-7-methoxy-2-(oxazol-5-yl) Quinoline synthesis yo 0 : 0

No Na 8 l-0 z a

A SP

OMEM OMEM wf ive b a

For only N N 0 « | N { x ~N = 0 | x | 0 free free OH OMEM cap 4 g a TOA solvent 11.8 g (mmol) of the buffered quinoline derivative as example; in 10 ml of 3:0 and cool the mixture to A VA= before adding 1/.5 ml diisobutylaluminum hydride (1 mol.mL) in toluene very slowly over YO min. After stirring for 81 minutes, add an additional 0.0 ml of 0,0 DIBAL equivalent; concentration 1.0 —u in (toluene) and after stirring at a VA= for 2 hours Two additions” The reaction mixture was watered carefully with § ml of methanol at A-L, then poured into a Sle solution of Rochelle salt tartrate (E16, concentration 1 M). The dough was stirred to become thick. thick paste using 01 ml of CHCl for 2 hours until it became clear.The phases were separated and the organic phase was dried over 0.04850, filtered and concentrated to produce a white solid.The result of purification by flash chromatography on SiO;the number of eyes ranges from From 710-5060 samples using a mixture of FtOAc/hexane of concentration 0, 8 g of aldehyde 5 1a as a white solid (production rate was IVY b. 1 mg added) ;?,; mmol) of toluenesulphonylmethylisocyanide to a stirred suspension of 58 174 piles (174 mmol) of 0.16.00 in 1 mL of ans MeOH reaction mix to vo©m and 0 was added 174 g (mmol) of aldehyde 41a. The reaction mixture was heated to 80 °C for 11 hours and then concentrated to dryness in a vacuum. The purification was carried out by flash chromatography on SiO, an Age number ranging from 0?7 ?-0 (po yield AY, + g of desired oxazole; Ab (production rate was (IA) a

Ya

MS: 331.0 (M+H)". Reaction for .0 M) 1 aqueous (HC) before concentration to dryness in a vacuum. The ore was treated with ©mL of pH buffer, pH 0.£ (and stirred before filtering the product; be) (solution of concentration of phosphate — © 0.068 g hr. VT was produced, washed with distilled water, and dried overnight in a vacuum at 0 °C for the desired duration; ?1 g as a yellowish-brown solid hydroxyquinoline from SEM) 77.4 MS (4) +0 (production rate is 'H NMR (DMSO-d): 5 8.65 (s, 1H), 8.02 (bs, 1H), 7.97 (d, J =8.9 Hz, 1H), 7.19 (5, 1H), 6.93 (d, J=7.9 Hz, 1H), 6.42 (bs, 1H), 3.87 (s, 3H).ES (+) MS: m/z 242.9 (M+H)". LODB NTMAM LAM LAHAK A L MMASAM NHAc A HOOC NHAc (ive) (0) NO va (e+ +/1) (5, S-B:-DUPHOS

BOH

(210) = NHAc psi”) 01 (with Hy-pressure 0 Pyridine Refrigerant for “i

B mm (ve) (=e) Ce) excess quantity ratio 30 > 14 (by (GC)

THF DMAP followed by 0 s, (1?) LioH. Mega | H. A 0

NHBoc on i

Coo 0) (=e) a. Canal 460 mL of aqueous sodium hydroxide (concentration 1 mole; 1 eq.) drop by drop to a solution of 100 g (7&5 mol) of IYO diethyl 2-acetamidomalonate commercially available at 0 ml of dioxane over a period of time ranging from yo to 1 minute. and the resulting mixture, oo, was left to stir for 11.00 hours and a half; Then dioxane was evaporated in a vacuum; The aqueous solution was extracted in three batches — ethyl acetate of 0 mL each aaa g to a pH of 1 using 1101 concentrate. And let this solution crystallize in a bath of water and ice. After the appearance of a few crystals; ale of the mixture for sound waves and an ample amount of precipitate appeared. The filtration and drying in vacuum resulted in 67,097 g of the compound (°Ab) in the form of a white solid (production rate 1 was 977/). b. An aqueous solution of 560.7 g (8 mol) of 11) sodium periodate equivalent was added; in EVO from 10 ml (to a magnetically stirred emulsion of Yo 0.1 thousand g mol) of commercially available 7-octene-1,2-diol 11g and 1100 ml of water in a 1 liter round bottom flask over a period of 10 minutes Jel (3 s ya exothermic Vo) 0. The resulting mixture was stirred at room temperature for one hour. Further (and prove the completion of the reaction with TLC). Then, the mixture was filtered into a separatory funnel.

The aqueous layer was separated from the organic phase. The aqueous solution was saturated with NaCl, filtered, and separated from the organic part again. The two organic parts were mixed, dried using sodium sulfate, and filtered onto a cotton plug (in a Pasteur pipette). 159,175 g of compound 210 was produced in the form of colorless oil (production rate was ZA) and the aqueous solution was extracted as Mb “CHCl”, dried using MgSO, anhydrous and concentrated in vacuum (without heating; using heptanal Boiling it 05" m) to yield an additional 1,907 g of NO as colorless oil (rate of production was 7 01). Total yield was mJ. EA added g (0 mm£) (Ue of heptenal 01D in a solution of YY ml of pyridine 0 equivalents) to Vor g (0£ mmol) of ethyl 2-acetamidomalonate 101b annealing through - One minute, the resulting solution was cooled in a bath with a temperature of 10 °C, and 11 ml of acetic anhydride (7.7 equiv.) was added within minutes.The resulting orange solution was stirred for ¥ hours at room temperature and another part of 1.77 was added. g of Vo ethyl 2-acetamidomalonate and the resulting mixture was stirred at room temperature for an additional 11 hours. Then 10 mL of ice was added and the solution was stirred for an hour and a half; Then dilute the mixture You yo mL of water and extract with two parts of .ether and wash the ether solution with HCl (concentration 1 M) and NaHCO;saturated; and dried over 20:50. Concentrated and purified by chromatography by chromatography using a mixture of EtOAc with a concentration of 400 7/UH”1 ph/,; 5g of the compound 5e in the form of a pale yellow oil (production rate was +10 (d). 0 ale of (S,9)-Et-DUPHOS Rh(COD)OTf was added (ratio = sulfur/carbon) 497) to vy. a degassed solution (forming argon bubbles for 70 min) of AYA g vo mmol) of Zethyl 2-acetamido-2,8- nonadienoate 1 h in 10 mL of dry ethanol. The mixture was placed in an atmosphere of hydrogen at a pressure of 701.8 kilopascal (KPa) ve (pounds/square inch (psi) (after; cycles of vacuum-addition 8 Yvy

And stir on a Parr shaker for two hours. The resulting mixture was evaporated to dryness to obtain the crude compound 0 (4) which was used in the next step without purification. e. 1.7 g of Boc (0 equivalent) Y (Vo mg +.Y) was added DMAP eq.) to a solution of 7.9 g (0,7" 0 mmol) of ala (S)-ethyl 2-acetamido-8-nonenoate at 310 M in 100 mL of THF and heated Toe reaction was carried out at reflux temperature for 2 ½ hours.Then most of the THF solvent was evaporated and the crude mixture was diluted with CHCl, washed with 1 M HCl to remove DMAP and the organic layer was further extracted with NaHCO; saturated aqueous, dried with 11:50 anhydrous and concentrated in vacuo.Then the crude product was diluted with 5+6 mL of THF and 0 mL of water; 7.54 g of 0.LIOHH (¥ equivalent) was added The resulting mixture was stirred at room temperature for 1 yo h (hydrolysis was confirmed by -TLC) and the reaction mixture was concentrated in vacuum to remove most of the THF solvent, diluted with CH,Cl, and washed the solution The product is 1 M HCI”, dried with 0.1,90 anhydrous and concentrated in a vacuum. To remove minor impurities and the excess amount of 0:800, pure the crude product by flash chromatography (using a mixture of a solvent with a concentration of ++) 7p EtOAc 700-1 as the fluidizing agent). 10 - 8.87 g of the compound labeled with heading 10 was produced in high purity in the form of a pale yellow oil (production rate was .)/971 Hz, Jd= 17.0, 6.7 NMR (DMSO, 400 MHz). : 5 7.01 (d, J= 8Hz, 1H), 5.79 (tdd, 11 Hz, 1H), 5.00 (md, Jd= 17.0 Hz, 1H), 4.93 (ind, Jd= 10.2 Hz, 1H), 3.83 (m, 1 H), 2.00 (q, J=10.2 Hz, 2H), 1.65-1.5 (m, 2H), 1.38 (s, 9H), 1.35-1.21 (m, 6H). 6.9 YYVY ‏

AY ive Example (J Vo ) (28)-N-Boc-amino non-8-enoic Acid TLP substitute synthesis

J

EA Sa L Dah Br hyo

JL Na L M :

HX «A

Nj ml | 5 dye D | d od OH

BocHN mm_|| BocHN

Ne dye

AY

Drop by drop into a stirred suspension of z 1 © 8-bromo-1-octene mmol) of 1 5) mL Y,0F a. Add dry THE mL of Vo cut to form Flour in Mg mmol) from YY bars, 0 gm 5

Tolls min; [The reaction was Vo over a period of time amounting to 1.1 ml of dibromoethane containing 1 hour and then cooled ad a minute;. Heat the reaction mixture to 78 °C Wo to slightly hot]. And yet on an additional amount of © , solid. The cannula M was diluted to L/- before adding it through a 50 ml cannula, and the solution was washed twice with brine water in the amount of 001 ml diethyl ether. Mixture with silica: by chromatography on a gel (JES) and evaporation. Crude oil was produced, MgSO, each time; it was dried over as the auxiliary material, producing 1.44 g of hexanes in its concentration of 11 EtOAc using a mixture of TY (the production rate was Vo compound 'H NMR (CDCl, 400 MHz) 8 1.31-1.42 (m, 6H), 1.60-1.69 (m, 2H), 2.02-2.00 (m, 2120, 0 2.35 (t, J= 8.3 Hz, 2H) , 4.99 (dm, J= 10.0 Hz, 1H),5.04(dm, J= 17.0 Hz, 1H), 5.75-5.86 (m, 1H), mL (3.04 mmol) of freshly distilled VV A EON (mmol) of 11.7 (1.1 ml) b. carboxylic acid (Use mM AY) was added to a vigorously stirred solution of 1.77 g anhydrous pivaloyl chloride at -4 L by means of a syringe under anhydrous conditions THF stirred ml of Ve in 0 °C for 0 min then at 0 °C for 0 ºC then cooled V0 for 0 VAS the mixture at os the mixture again to -8L and then transferred via a cannula to an anhydrous solution of lithium salt. A VA- salt was prepared at THF (8 4-(S)-4-(phenylmethyl)-2-oxazolidinone lithium (Use mM 10,Y) was previously prepared by the slow addition of 7.88 mL oxazolidinone to the reactant g 0.786 g THF 0.786 mL of Yo to a solution in (molecular hexanes in Y (concentration n-BuLi from -/L. The reaction mixture was stirred at -8/m for oxazolidinone (mmol) from Vor © min, then at room temperature for an hour and a half. Finally, it was irrigated with an aqueous solution from 1 to 7/;_ its initial volume. THF molecular) and evaporated 1 (its concentration sodium bisulfate ml of 0 ml each time, and the organic layers were washed after 151 by BIOAC And the ore was extracted twice b

He mixed it three times with NaHCO of 70 by 50 ml each time. and twice with brine water of 50 ml each time; dried over MESO, and evaporated. The resulting crude oil was sorbed on a csilica gel using EtOAc at a concentration of 710 in hexanes to obtain 1.88 g of the compound (sho). The production rate was ATA m 'H NMR (CDCl;, 400 MHz) § 1.35-1.47(m, 6H), 1.67-1.74 (m, 2H), 2.02-2.09 (m, 2H), (dd, J= 13.4 & 9.9 Hz, 1H), 2.84-3.02 (m , 2H), 3.31 (dd, J=13.4 & 3.2 Hz, 1H), 4.13-4.22 2.65 (m, 2H), 4.62-4.71 (m, 1H), 4.93 (d, J= 10.2 Hz, 1H) , 5.00 (dd, J=17.2 & 1.6 Hz, 1H), 5.75-5.84 (m, 1H), 7.18-7.38 (m, SH). c. JE solution of FYO g V0) mmol) of the acid derivative (B50) 56 mL of THF© dried at VA-2 via a cannula to a stirred solution of YY mL (17.5 mmol) of 8 (concentration + Molecular A) in *0 mL of dry THF at -4 L. The mixture was stirred at VA for 0 £ min. To this mixture a solution of 3.17 g (11.85 mmol) was added of the building in 0?ml of The dry at -4/Lum.The mixture was stirred at -4L for 7 minutes, then irrigated with ©ml of acid 6N2061.Then stirred at room temperature for one hour and 45 minutes yo Finally at 0 µm for 10 min. Most of the THF was evaporated and the concentrate was dissolved in 100 ml of EtOAc and the organic solution was washed with *0 ml of (HL0) times with NaHCO; Its concentration is 75 by 0 mL in every 5350 00 mL of brine water; And dried over ,14850 and evaporated. The resulting oil was absorbed on a silica gel using a mixture of hexane/CH,Cl, (at a ratio of 01) as a thinner, resulting in 1 kg of the compound, VV, Jane. The production reached (NY (m, 6H), 1.45-1.6 (m, 1H), 1.75-1.88 (2, 2H, + 1.32-1.45 H NMR (CDCls, 400 MHz) rotamers), 2.01-2.11 (m, 2H), 2.82-2.87 (m, 1H), 3.33 (dd, J= 13.4 & 3.2 Hz, 1H), 4.10-4.28 (m, 2H), 4.62-4.72 (m, 1H), 4.90-5.05 (m, 3H ), 5.73-5.88 (m, 1H), 7.17-7.38 (m, 5H). lodged in

Ao (mmol VFA) 7.11 g via a cannula to a stirred solution of SLY MeOH mL of anhydrous Yo. The mixture was stirred at room temperature MeOH ml of Ae from anhydrous ©5800 in the resulting mixture of foamy material and the foamy material MeOH was dissolved for a period of time; hours. (mmol) of VTA) 1 μl of water was evaporated and treated with © 71 gm, and 001 μl of dioxane. The pH was adjusted until Cua) NaHCO; g (5,“ mmol) of Y,A% 5 800 0 as needed) and stirred at room temperature NaHCO; By using more A EtOAc, the ore was extracted twice with dioxane an hour. Evaporation of a part of about 70 out of 11 for a period of time and washing the organic solution twice with brackish water in the amount of 56 ml each time; It was dried and evaporated. Its hexane concentration was graded in EtOAc using a mixture of silica and the resulting concentrate was sorbed on a gel (1). “HNMR (CDCl, 400 MHz) 5 1.27-1.53 (m, 6H), 1.46 (s, 9H), 1.8 (m, 1H), 2.00-2.08 (m, 1H), 2.8 (t, J= 12.1 Hz, 1H), 3.34 (d, J= 14.3 Hz, 1H), 4.17-4.23 (m, 2H), 4.60-4.66 (m, 1H), 4.93 (d, J=10.2 Hz, 1H), 5.05 (dd , J= 17.2 & 1.9 Hz, 1H), 5.13 ( bs, 1H), 5.38-5.43 (m, 1H), 5.74-5.84 (m, 1H), 7.22-7.36 (m, 5H). 0 (in proportion HO; of (Use mVLY) mL Yeo e. 6 was added to a stirred solution at 0°C of 1.74 g (.04 mmol) of LIOHH,0 mmol) of AY) mL of 11:0 and stirred the solution at 15°C and THF 15 mL of VO in a mixture of L100#300-17 derivative 11:50, Use 17.8 mL (15 mL 15 0 C) for one hour. The reaction mixture was irrigated with aqueous citric acid, pH between (da) and ©, and adjusted (ele g in YY)

EtOAc, the second part was extracted again with EtOAc, the mixture was diluted with 701 (concentration 0 silica), the organic solution was washed twice with brackish water, dried and evaporated. In 208 its hexane concentration using a mixture of production amounted to 7). This compound was identical in all Jaa) Ye talc carboxylic acid

AY

(Ye) aspects of that obtained in Example 11 Example D(1) (28)-N-Boc-amino-5-oxo-non-8-enoic Acid Methyl Ester Synthesis of BocHN, __00CHs Mg | OC 0A / Bee #Lahtia B — me i

A oom Aor C - BocHN._CO:CHy i Sr Hn and his collaborators in the journal T.Tsuda This synthesis depends on the method of T. Tsuda °

AYA AYAE=TIYAY p: 00 vol. J Am. Chem. Soc point by point hexane (concentration of 59+ moles in n-BuMg from (Use m©,Y) a 5.8 ml of malonic acid monoallyl ester core (mmol) of VE was added to a well-stirred solution of 1.0 g ml at -m/cam during a time period of ~ minutes. Then stir 10 by N, dry in an atmosphere of 7 thick suspension at room temperature for one hour and evaporate to dryness (release 1 steel in a vacuum for one hour. Mg and dry with 0 salt). (N, vacuum in an atmosphere of mmol) of 0171) first with 1.1 g 111 glutamic acid and an anhydrous derivative was mixed and the mixture was stirred at room temperature for an hour THF at 1.1 '-carbonyldiimidazole

the mom

One to activate the free acid moiety, and later the activated glutamic acid derivative was transferred.

Through a cannula into a solution of Mg salt 11b and stirred the resulting reaction mixture at temperature

room for 6 hours. Then it was diluted with 2086 and the organic solution was washed with HCl and cooled with ice

(concentration, 0 M); brine water; Then it was dried and steamed. The resulting concentrate was sorbed on a silica gel using a mixture of EtOAc in hexane with a concentration ranging from 18-759 as a liquefied material.

1g of compound 11g (production rate was 70%)

'H NMR (CDCl, 400 MHz) 8 1.44 (s, 9H), 1.85-1.95 (m, 1H), 2.12-2.22 (m, 1H), -2.58

2.74 (m, 2H), 3.48 (5, 2H), 3.74 (s, 3H), 4.24-4.34 (m, 1H), 4.52 (dm, J= 5.7 Hz, 2H), 5.09 (m,

1H), 5.25 (dm, J=10.2 Hz, 1H), 5.34 (dm, J= 17.2 Hz, 1H), 5.91 (m, 1H).

0 b. TAY g (¥ mmol) of diester was added 71 g in mL of dry DMF (by

dy diss atmosphere) to a stirred solution of 6.116 g (0 mmol) of

tetrakis(triphenylphosphine) Pd (0) (at a ratio of 78 mol) in 1 mL of dry DMF. And stir the mix

at room temperature for Glebe and a half. DMF was evaporated under reduced pressure and diluted

the ore with 10 mL of 2:08 and a solution of EtOAc washed with © mL of cooled HCI 1,0N gill and 10 mL of brine; And dried and evaporated. And the concentrate was absorbed on silica gel using

A mixture of EtOAc in hexane with a concentration of 7701-11 as a thinner, yield 0.107 g

of compound 10d (production rate was 47/).

'H NMR (CDCls, 400 MHz) 1.44 9H), 1.84-1.94 (m, 1H), 2.08-2.22 (m, 1H), 2.33

‎(dd, J= 14.0 & 7.3 Hz, 2H), 2.45-2.55 (m, 4H), 3.74 (s, 3H), 4.28 (bm, 1H), 4.98 (dm, J= 10.2 Hz, 1H), 5.03 (dm, J= 17.2 Hz, 1H), 5.00-5.10 (m, 1H), 5.74-5.85 (m, 1H). Ye

a

AA

Ex. 11 (and 11) (28,5R)-N-Boc-2-amino-5-methyl-non-8-enoic NO 3A.NGAD synthesis *NHAC +-BuOH/ H;0/scotons Ch ia m m : - = i

COR OH COLE

CH; ivy wy lag <r THF/H,0 Bac0 (1) Ph3PCH Br (1)

THR DMAP CH, KHMDS CH,

SC, =e Cr, Jk Cr cos © cog 2 0 rum NY uh NY to the available Glas amino acid derivative IVY R(+)citronellal I Joa a b c; d. aldehyde to convert Yo by following the same synthesis steps previously described in example B1 amino acid ° +3) © amino acid to the intermediate compound of the amino acid (310) ml in a mixture of 18 V QIY of the compound (de Ge 0.1) g 1.175 e. dissolved (at a ratio of 1:1) and placed in an ice bath at 0°C. tBuOH/acetone/H,0O mL (+50 mmol) of 17 eq) and NMMO, (1 g) were added 74 mmol) of 4. tig reaction mixture (equivalent) in a series of 1,017 tBuOH (at a ratio of 77.8 w/w in 0.090 by evaporating in a vacuum and then extracting the acetone at room temperature for hours Most of the mixture was removed with 6 M©8 The organic layer was additionally washed with water and brine and dried over high purity after 11g diol g of anhydrous 0V0+ and evaporated to dryness yielding 10 + as EtOAc in 1(7) (concentration EtOH) by flash column chromatography using a mixture of lysate (production rate in g OVO mmol) of ,11810 (,7 eq. ) to a solution of Y,Y0) g 8A W. was added

5 M (177 m) (Use of 11g diol in 01 mL of a mixture of 0.0 THF/H (1:1 ratio) at 0 °C and stir the reaction mixture at room temperature Most of the solvent Ga THF was removed by evaporation in vacuo and the remaining mixture was extracted twice with 21086 by 0 mL each time The organic layers were further mixed twice with © 70 by 01 aqueous cetricacid solution mL each time”; 01 mL of NaHCO;Mathe 75 and twice with brine water at a volume of 50 mL each time then the EtOAc solution was dried over anhydrous 14850, and evaporated to dryness in a vacuum. Use 0, 47 g of an intermediate of 11d aldehyde as the crude product in the next step without further purification g.8.1 mL (7.1 mmol) of KHMDS (its molecular concentration in (toluene) was added to a solution of 0.0 mg AYO (7.1 mmol) of :11:7011:3 in VO ml of anhydrous toluene and stirred the yellow suspension formed at room temperature for Te min at atmosphere of Ny and after this period; 5) the suspension was first brought to zero degrees Celsius; and a solution of 47 g (1.77 mmol) of VV aldehyde (dissolved in 11 ml of anhydrous THF) was added via syringe and let the Bad mix room temperature. After stirring at room temperature for 1 hour, most of the THF vo was removed by evaporation in vacuum; 100 ml of 08:5 was added to the mixture and the organic layer was washed with 01 ml of 0.0 1 ml NaHCO;70 aqueous and 00 ml of ele brine. Then the EtOAc solution was dried over MgSO, anhydrous and evaporated to dryness in a vacuum. 179 g of the pure compound 11H was separated after purification by flash chromatography column on a silica gel using a mixture of EtOAc s hexane at a ratio of 7:7 as the lytic (production rate was 777 for the last two steps). x. The hydrolysis of the ethyl ester core and simultaneous substitution of the N-acetyl protective group of Boe in intermediate compound 1e were carried out to obtain ada 701 from compound 71f (quantitative production rate) using the same aforementioned procedure to convert Compound 51f to *1g

(d, J= 6.4 Hz, 3H), 1.18-1.28 (m, 2H), 1.35-1.48 (m, 0.88 E' H NMR (CDCl;, 400 MHz): 3H), 1.45 (s , 9H), 1.64-1.74(m, 1H), 1.81-1.89 (m, 1H), 1.94-2.12 (m, 2H), 4.28 (bd, J= ~3.2 Hz, 1H), 4.93 (dm, J= 11.1 Hz, 1H), 5.00 (dm, J= 16.8 Hz, 1H), 5.74-5.84 (m, 1H). Example VA °A) N-Boc-O-allyl-(L) synthesis (-threonine 99) OH 0 L 0 OH B i : A — M A NHBoc NHBoc ab tA J J 0 0 0 0 — AA - A NHBoc NHBoc dg A. ©0000 mg Y,YA (mmol) of VA Boc-(L)-threonine were partially dissolved in a mixture of A mL of CHCl, and * ,1_ ml of (MeOH) at zero degrees Celsius. A solution of diazomethane in diethyl ether was added slowly until the color “pial” was fixed, which indicates the presence of an excess amount of Ye of diazomethane, and by evaporating the solvents, 4 6.07 g of crude methyl ester VA in the form of cloudy white oil b.Then 011 mg (1.77 mmol) of intermediate CVA was dissolved in A ml of diethyl ether anhydrous, and 41 mg (7?.1 mmol) of AZO and 1 g of Boas activated molecular sieves of diameter; angstrom units were added. Finally, 194 µL (VV mmol) of allyl iodide 0 to the reaction flask ily the mixture at the reflux temperature. I added two IVY parts

two additions of allyl iodide of £0 μl (2.0 mmol) each after a period of time

71 hours 10 hours; The stirring was continued for a total time of ½ hour. Then filter the mixture

During silite and purification by flash vertical chromatography on a 501:66 gel using a mixture of

EtOAc/hexane in a ratio of 1: as a fluidizing agent; VY produced 71 mg of the compound in the form of a clear oil (the production rate was 771).

'H NMR RL (0i) 400 MHz): 1.21 (d, J= 6.0 Hz, 3H), 1.45 (s, 9H), 3.75 (s, 3H), -3.82

3.87 (m, 1H), 3.99-4.07 (m, 2H), 4.29 (dd, J= 9.5 & 2.5 Hz, 1H), 5.14 (dm, J= 10.5 Hz, 1H), 5.21

(dm, J= 17.2 Hz, 1H), 5.75-5.84 (m, 1H).

c. Dissolve 94 mg (TTY) 0.0 mL (Use) of ester compound 81 g in 4 mL of a mixture of 0 1170/01771.0” in a ratio of 1:0: 11 mg V, € added 0) milli (Use from 1101111:0.lis

the solution at room temperature for 2 hours; Then (ala using HOI (concentration 1 M) to

The pH was reached about before solvent removal in a vacuum. I have used the oil compound

Product 2)1A as is for the synthesis of -macrocyclic inhibitors ay 19 Ex. (= 14 ) (28,3S)-N-Boc-2-amino-3(Mercaptoallyl) butanoic Acid Synthesis of 0 a ah { en A de mo “roh | ox a free from ine A, Jo jezini” 17 its concentration is 1 2 NaOH Reap 0 except LER for

Ne 1 min we = N TN ground solution ds pyridine mmol of compound 811 (©mL of 9.1 a.d. solvent in equiv.) 1,7) tosyl chloride mmol (of 11.8) C in an ice bath; “,7 g” was added 0 h. After this period; YE was added in few batches and the reaction mixture was stirred at room temperature for 11.0 ml ether. The layer was washed 001 and diethyl ether mL of Yoo Distribute the reaction mixture between 001 mL each and 001 N of 17 N HCl additionally + times anhydrous b; filtered and concentrated until MgSO was purified and dried over a glass of water (hexane/BIOAC concentration gradient) the raw material by flash column chromatography, using a mixture of -0AP (tosyl modifier as a Isolate ¥ 0.00 g of a :17 to 0:8 derivative (IAS yield in equivalent) to a solution of 1,1) potassium thioacetate B. 15 mg (7.7 mM) added mol) of anhydrous DMF and stir 0 ml of Y.0 of the intermediate compound 1911b in (Ue of 5 mg) ¥ mL in vacuo and DMF dispense the reaction mixture at room temperature for 7 hours; Then most of Ne

Yvy

The remaining mixture is between BtOAC and water. The aqueous layer was re-extracted with EtOAc and the organic layers were washed after mixing with brine; It was dried over anhydrous 1/850 and evaporated to dryness. Purification of the raw material by flash column chromatography using a mixture of hexane/EtOAc (at a ratio of 6:1) as the lysing agent resulted in the isolation of 10 £pile of compound 91g (production rate of (IA)

0 c. 7.4 mL of an aqueous solution of 1.7 mole NaOH was added to a solution of 8 mg of 91g thioester in A mL of a mixture of 11.0/21011 (0:0 ratio) and stirred Mix at room temperature for 1 1/2 hours Then 797 mL (7, mmol) of (Y) allyl iodide equivalent) was added and stirred at room temperature for an additional 710 minutes The Jeli mixture was concentrated to half its original volume Then extracted with B 8086. The aqueous layer was reduced to

approximate pH? using cold-pressed aqueous HOI of 1.5 N and re-extracted at Am 80. The organic layers were washed after mixing them with brackish water; It was dried over MgSO, anhydrous, and evaporated to dryness in a vacuum. The crude reaction mix contained; products at least; All products were isolated after flash column chromatography on a csilica gel using a mixture of hexane/EtOAc (4:0 to V7). The structure corresponded to the less polar compound (as specified).

ve TLC VA in a mixture of hexane/EtOAc at a ratio of AY (0:4 mg of desired product 4d) yield was A (7) 'H NMR (CDCls, 400 MHz: 5 1.24 (d, J= 7.0 Hz, 3H), 1.46 (s, 9H), 3.13-3.19 (m, 2H), (m, 1H), 3.77 (s, 3H), 4.50 (dd , J= 8.6 & 3.8 Hz, 1H), 5.12 (d, J= 12.4 Hz, 1H), 5.15 3.24-3.29 (dd, J=18.4 & 1.3 Hz, 1H), 5.22 (bd, J= 7.6 Hz , 1H), 5.75-5.85 (m, 1H).

3 0 d. Mix a solution of AY mg YAY) mM (J se of methyl ester 91 d in ; mL of the mixture

of 0.0 MeOH/H (ratio: 1) with VF 1771 ml (mmol) of aqueous NaOH concentration

0" titer for ;7 hours at room temperature and for 1 hour at a fr and acid

The reaction mixture was cold aqueous HCL (concentration 0.5 N at zero degrees Celsius; −° in vacuum and the remaining aqueous mixture MeOH was extracted, its acidity ranged from − to ©) and was removed and evaporated to dryness to obtain the compound MgSO, and the organic solution was dried over EtOAC B 4e. Compound 1e was used for the final synthesis of the inhibitors without any subsequent purification. 2-amino-3-methyl-3 (1-mercapto-4-butenyl) butanoic Acid Synthesis ° ~~ ~~ 8 !7 wn

I QOH (° Sahi —- Sahi ir. ANN wre at.

Boo, O) 1

DMF/DMSO 1 mL of a mixture of 1 in A01 Lopenicillamine prof. Dissolve 448 mg (¥ mmol) of 4-bromopentene (1,0 eq) (in a 0:1 ratio) and add 47 ml (*, 5 mmol) eq) and stir the reaction mixture at After 0.111.0 g (+ml) of 10u eq) to the mixture and continue to mix (Use 0.7:0.7) of mg (7.75 mL 871 h) was added. YE.

HCl and the remaining mixture was diluted with cel gd in Ga ¥ DMF an additional hour. VY was removed by stirring for a titer period) and its pH was adjusted between about ; Then, 1.5 cold aqueous was extracted twice (concentration ml each time). The organic layer was washed twice with brine water and dried over 50 B 20/6 by 0 B 10 crude anhydrous carboxylic acid and evaporated. B. The purification of compound 10b proved difficult, so the crude product was first treated, Vo, and then purified by flash column chromatography, using the corresponding ae methyl ester to form an amalgam of methyl ester. 0980 (in a ratio of 1:9) as a liquefied material; a mixture of pure 718/0g hexane/BtOAC was produced (the production rate was only 0.7g).

0 'H NMR (400 MHz, CDCL): 8 1.35 (s, 3H), 1.37 (s, 3H), 1.44 (s, 9H), 1.59-1.67 (m, 2H), (m, 2H) , 2.51-2.60 (m, 2H), 3.74 (s, 3H), 4.29 (d, J= 8.6 Hz, 1H), 4.98 (dm, J= 10.5 Hz, 2.11-2.17 1H), 5.03 (dm, J=19 Hz, 1H), 5.35 (bd, J=7 Hz, 1H), 5.72-5.83 (m, 1H). The ester was subsequently dissolved in ¾ mL of a mixture of THF/MeOHH,0 (at a ratio of 7:7:0); 00 mg (7.0 mmol) of Y (LIOHH,0 eq) was added and the mixture stirred The reaction at f+m for a duration of ; hours to hydrolyze (—= Y 0 ) ester back to acid ) 0 b . The reaction mixture, HCl, reduced its concentration to 1.5 N to a pH between ; and ©; THF and Jia 146011 evaporated dryness and extracted the remaining aqueous solution — EtOAc. The layer of EtOAc was dried over MgSO, anhydrous; It was evaporated to dryness, yielding the compound Ye, which was used in the subsequent synthesis of massive Ve cyclic inhibitors without postremediation. Acyclic Dipeptide and Tripeptide iu Intermediates Description of the general procedure for in-solution bonding reactions and specific examples in Jel pn Intl. No. 09847/010 and No. ally . 048048/01 These B procedures were used for the synthesis of intermediate compounds from the intermediate dipeptides iu Hg 01 and the tripeptide compounds Cat Kc “A YY PY Packer

Example YY Synthesis of Acyclic Tripeptide (11E) 0 N Fy J 0 DTM O MACS OH OLA % OA N 0 \ Fr 0 1 0 ivy | (0 0 mmol) of 1.0F 5 NMM mL (£507 mmol) of HATU successively to a solution of 1.97 g TTA (mmol) of a proline (IY) derivative (prepared from the commercially available 4-chloro-quinoline 5 Boc-4(R)-hydroxyproline as described in International Patents No. 101 and 4808 (ref) and approximately ¥0,¥ mmol of homoallyl Crude ACCA or in 0 ml of CHC, and the suspension was stirred at room temperature for 1 hr. After this period, the solvent was evaporated and the crude reaction mixture was re-dissolved in 0 ml of YA. EtOAc and washing Ava av ml of 10 ml water and 10 aqueous B with a concentration of 70 in an amount ranging from ? to NaHCO; solution B using silica mixture and evaporation.The crude product was purified by chromatography on a MgSO gel, brine; and dried over as a liquefaction material to obtain the stereoisomer EtOAc with a concentration of 78 in diethyl ether from (the stereochemistry did not specify) 0 of the desired compound 1"b (the production rate was diastereomer. (absolute stereochemistry ° 'H NMR (CDCl, 400 MHz): 0.93 & 1.01 (t, J= 8.3 Hz, 1H, rotamers in ratio of 3:7), 1.14-1.35 (m, 2H) , 1.44 (s, 9H), 1.45 (s, 9H), 1.50-1.82 (m, 4H), 2.08-2.24 (m, 2H), 2.32 (bs, 0.7H), 2.63 (bs, 0.75H), 2.93 (bs, 0.75 H), 3.16 (m, 0.25H), 3.77 (bs, 1.5H), 3.88 (bs, 0.5H), 4.4- 4.55 (m, 1H), 4.98 (d, J=10.2 Hz, 1H ), 5.03 (dd, J= 17.2 Hz & 1.6 Hz, 1H), 5.24 (bs, 1H), 5.75- 5.88 (m, 1H), 6.57 & 6.78 (2bs, 1H, 2 rotamers), 7.42-7.58 (m, 3H), 7.63-7.73 (m, 2H), 8.04 (d, - 1- 8.3 Hz, 1H), 8.11 (d, J= 8.3 Hz, 1H), 8.74 (d, J=5.1 Hz, 1H). ) of a dipeptide (+, YEA) and a half. Then the solvent was evaporated and the concentrate was dried in extreme vacuum, resulting in delamination at room temperature for a period of talc diethyl ether. The mixture was dissolved in a mixture of ¥ μl of amino acid Vo dissolved in diazomethane and treated with a slightly excess amount of MeOH μl of Yo

HCl By adding diazomethane, the excess amount of diethyl ether was destroyed and the mixture was evaporated to dryness to obtain the 1101 salt of the compound (dioxane) (its concentration; molecular V) which was used in the next step without any purification. C 71 “3Y) (28)-N-Boc-amino-hept-6-enoic acid g (17,” mmol) of 0.101 g vy added to HATU (17©) mmol) of 1777 NMM and 1.91 mmol (1.91 mmol) of crude 0 dipeptide (mmol) of dipeptide +, EA) g YY cascading into a stirred suspension of 1 h 011 and stirred the mixture at room temperature for CHCl, mL of YO in 11C aA

Gua after one hour using NMM A (the pH was adjusted to about and the organic solution was washed twice with EtOAc as needed). and evaded you 011:0; and dissolve the ore in 00 ml of ml in each Yo ml each time; and twice with brine water in an amount of 100 with a concentration of 70 by NaHCO, b (using 00 ml of a mixture of silica, dried and evaporated. The resulting crude compound was absorbed on a 63 ye gel (7870 g of the compound 17 H) (the production rate was 1.179 vintage (BOAC//Y of 80 concentration © 'H NMR, Y02) 400 MHz, rotamers in 1 ratio) chemical shift of major rotamer: & 1.21-1.27 (m, 1H), 1.36 (s, 9H), 1.45-1.81 (4m, 7H), 2.20-2.22 (m, 4H), 2.28-2.37 (m, 1H), 2.90- 2.99 (m, 1H), 3.66 (s, 3H), 3.94 -3.98 (m, 1H), 4.29 (bd, J= 9.9 Hz, 1H), 4.46-4.50 (m, 1H), 4.81 (dd, J= 8.3 & 5.4 Hz, 1H), 4.92-5.06 (m, 4H ), 5.16 (d, J= 8.3 Hz, 1H), 5.37 (m, 1H), 5.70-5.84 (m, 2H), 6.82 (d, J= 5.1 Hz, 1H), 7.47-7.55 (m, 2H) , 7.71 (dt, J= 7.0 & 1.3 Hz, 1H), 8.03 (d, J= 1.8.6 Hz, 1H), 8.17 (d, J=8.0 Hz, 1H), 8.78 (d, J= 5.1 Hz, 1H).

Macrocyclic Peptides Macrocyclic Peptides

YY example by macrocyclization the general procedure for the formation of massive rings olefins — metathesis Vo with a concentration of CHCl in tri-peptide diene in all cases; Dissolve tripeptide diene (for 1 hour argon from solution by forming bubbles of molecular oxygen and remove 61 171 ml). A solution of the catalyst was added at a rate ranging from approximately 0800 to 0800 until a volume of degassed was obtained, and the reaction mixture was heated with reflux until CHCl (dissolved in a small amount of HPLC and HPLC mixtures) was concentrated. TLC all feedstock to the product(s) as determined by means of a © ssilica short gel pad until almost dry and filtered through a Gay reaction crude wad to render all bulky cyclic product EtOAc B,011: 01 to remove most of the catalyst then by lysing Yl (bulk ring products) (found most of the time as a single stereoisomer).

Raw (Products) of each Jeli by non-chiral HPLC on a CHIRALCEL column Oh Jn 1 e CHIRALCEL (obtained from Chiral Technologies Inc) Diameter +87 mm, Length (aw YO using a mixture of isocratic solvents of 7970 11.0 + a mixture of CH,CN 7 + 5 TFA 7 005 + .178-70.05 at YO nm.m0 and characterize the main bulky cyclic product ( major macrocyclic products) completely by RN data “TOCSY” COSY ¢'H and ROESY to confirm its structure and stereochemistry. Example YY Synthesis of Macrocyclic Intermediate "B OH 0 Pd —r 8 th d" = 0 Li SC - except Ivy OH 0 8 f ~ 0 / sweeter 0 “Ty 1 Oxygen is removed from a solution of (g (VIA) mmol) of (BIYY diene) 800 ml

Veo

Ar by forming bubbles of dry (anhydrous) Aldrich obtained from Aldrich Company CHC, (by Hoveyda's catalyst of Hoveda's catalyst (se) for two hours. Then 767 g (471 mL) was added and after Ar b Lae as a solid and the reaction mixture was heated with reflux in a balloon (Use 70.0 h; the reddish-orange solution was evaporated to an amorphous solid and then purified by YA flash chromatography and the primary solvent system was silica over Gel from flash column column chromatography ° and once the catalyst is liquefied from .CH,CL, in 01% EtOAc, it is pure initial solvent system. EtOAc column; the solvent turned into colorless foam in the form of a colorless foam VY macrocyclic product The bulky cyclic product resulted from the evaporation of the solvent 0 (Y A) (with a ratio of about CH,Cly/hexane) It was re-dissolved in a mixture of .)/849 g white powder (yield 7 ye 'H NMR (CDCls, 400 MHz): 1.2-1.5 (m, 6H), 1.43 (s, OH), 1.53 (dd , J=9.5 & 5.4 Hz, 1H), 1.61-1.70 (m, 1H), 1.76-1.9 (m, 2H), 2.05-2.26 (m, 4H), 2.45 (d, J= 14.3 Hz, 1H), 3.67 (s, 3H), 3.71 (d, J= 11.1 Hz, 1H), 3.9 (dd, J= 11.1 & 4.3 Hz, 1H), 4.43-4.53 (m, 2H), 4.76 (d, 1-6

Hz, 1H), 4.86 (bd, J= 9.8 Hz, 1H), 5.2-5.23 (m, 2H), 5.57 (dt, J=7 & 9.8 Hz, 1H), 7.32 (bs, 1H).

011

Ye Example (£) Macrocyclic Intermediate Synthesis of Macrocyclic Intermediate 0

CO

{ hyd bar 0 *

Ye © “ Ng OMe 1 E 0 ° > Ab ml 001 BYE diene from (Use of a solution of 7.76 g (7.87 mm oxygen) removed mg 117 hours and ½ ad Ar of 1.0 anhydrous was added by forming bubbles of ©

CHCl, eq.) in + mL of +, + 0) Hoveyda's catalyst (mmol) of Hoveyda's catalyst +519) and the reaction mixture was stirred at the reflux temperature in an anhydrous and degassed cannula using a cannula with Als added. pal approximately; And at this 75 0 hours the reaction is completed by 71 and after Ar b Lina an additional hour balloon. Then he concentrated 011 for dail) a second part of the amount of 1 gram of the catalyst and continued 011 cm of silica gel pad and placed on top of a filling ¢ Ja Yao the solution to about 1

011 and the catalyst was first extracted by electrophoresis using ,01:.01. and washing the compound; QV was obtained from a silica filler using MeOH (at a ratio of 77) in 10/6 and purified by chromatography using a mixture of EtOAc/hexane in a ratio of 0:7 yielding 1.85 g of solid ale. White, slightly olive in color, Tan (with a purity of 744 as determined by HPLC and a yield of 97/6). , 1 9.2 Hz, 1H), 7.5-7.44 (m, 2H), m 7.17 (dd, J= 9.2, 2.2 Hz, 1H), 7.04 (d, J= 6.4 Hz, 1H), 5.6-5.56 (m, 1H), 5.52 (dd, J=9.2 Hz, 1H), 5.25 (dd, J= 9.2 Hz, 1H), 4.59 (d, J= 11 Hz, 1H), 4.44 (dd, J= 9.2 Hz , 1H), 4.05-3.98 (m, 1H), 3.94 (s, 3H), 3.92 (5, 3H), 3.89-3.82 (m, 1H), 3.55 (s, 3H), 2.64-2.53 (m, 1H) ), 2.46 (d, J= 7.3 Hz, 1H), 2.4-2.31 (m, 1H), 2.21 (dd, J= 8.9 Hz, 1H), 1.78-1.65 (m, 2H), 1.55 (dd, J= 4.8 Hz, 1H), 1.485 (dd, J= 4.8 Hz, 1H), 1.41-1.3 (m, 7H), 1.16 (s, 9H). Ve

MS; es”: 795.4 (M+H)”. A yoy

Yo Example (Y) Synthesis of compounds 707 and 1707 (Table

N N

& l 5 cm abh 8 b te

N i N

J 9 > 0 Barid Lah Touma 7 AH = iyo

N

& . 1 5 N 8

N N

> 0

AEN”ey: . 7 Compound YX Compound Is Y (diene g) in millimoles) of compound 1 a. A ring of mmol) of LI 1¢) amalgam oy was formed using catalysts of ° “Grubb's catalyst (Western bis (tricyclohexylphosphine) benzylidene ruthenium IV dichloride) with heating at reflux temperature for 2 hours. Produce CHCl, ml of 0 in the said Gla (ml of a mixture of 0 (using silica mg of compound 7) after chromatography on a gel of 1 (7 EA) (production rate was VAS EtOH/EtOAC 'H NMR ratio (CDCls, 400 MHz): § 1.22-1.30 (m, 2H), 1.35 (s, 9H), 1.44-2.35 (m, 13H), 3.07- 000 3.14 & 3.16-3.24 (2m, 1H, rotamers in 1:3 ratio), 3.69 (s, 3H), 3.96-4.04 (m, 1 0H), 4.42-4.50 (m, 1H), 4.95-5.04 (m, 1H), 5.05- 5.15 (m, 1H), 5.2-5.3 (m, 1H), 5.55-5.65 (m, 1H), 6.75-6.79 (2d, J=

Vet 5.4 Hz, 1H, rotamers in 1:3 ratio), 7.36 (s, 1H), 7.46-7.50 (m, 1H), 8.03 (d, J= 8.3 Hz, 1H), 8.13 & 8.17 (2d, J= 8 Hz, 1H, rotamers in 1:3 ratio), 8.77 (d, J= 5.1 Hz, 1H). mmol) of the bulky cyclic compound (0.0 YT) g 1015071 ester b. Dilute a molar fraction (mmol) of 11011.11:0 in a mixture of YT (mg AY A) using 8 macrocyclic compound inverse HPLC and 7 mL of 11:0. The crude product was purified with MeOH and 7 mL of THF from 4 mL of © (Whatman type) on a C18 reversed phase HPLC 08 C phase with a concentration of ude cm and diameter 7.4 cm using 50 long Partisil 10.0DS3 Partisil photo AVY all mg of compound 18 of 011.077 ph 001 to = CHiCN of a white amorphous solid. H NMR (DMSO, 400 MHz): § 1.12 (s, 9H), 1.2-1.24 (m, 2H), 1.32-1.40 (m, 3H), 1.58- 0 1.62 (m, 2H), 1.68-1.78 (m , 3H), 1.95-2.02 (m, 1H), 2.08-2.18 (m, 2H), 2.40-2.59 (m, 2H), 3.97- 4.00 (bd, J=9.8 Hz, 2H), 4.47 (t, J = 8.6 Hz, 1H), 4.58 (d, p 11.8 Hz, 1H), 5.22-5.29 (m, 1H), 5.46-5.54 (m, 1H), 5.66 (s, 1H), 7.12 (d, J = 6 Hz, 1H), 7.49 (d, J= 3.5 Hz, 1H), 7.68 (dd, J= 7.3

Hz, 1H), 7.98 (dd, J= 7112, 1H), 8.08 (d, J= 8.3 Hz, 1H), 8.21 (s, 1H), 8.35 (d, J= 8.3 Hz, 1H), 9.08 (d , J=5 Hz, 1H). Vo io macrocyclic compound (mmol) of the macrocyclic compound 0 FY) mg 10 c. Stir ml of dry .011:01 in the presence of © ml of a mixture of pH:1101/0 (molecular concentration) for 1 in ml of CHCl and 1 for 1 hour. Evaporate the mixture and dry it carefully. The ore was re-dissolved in ¥ ml of (mM YT) nor μl NMM (mmol) of 1177 (1.59 μl) and treated for 7 hours. The mixture was evaporated and dried 16 ad and stirred at 16 °C. room temperature acetic anhydride (mol) of ©, mL of MeOH, and 7 mL of THF in vigorous vacuum. Then the ore was dissolved in a mixture of; mL of mmol) from 1:011.211,0. The ore was purified (+, ¥T8) to 11 mg and stirred overnight with isolated H,0 N1 after acidification to pH - ¥ using ice-cooled 1101 L1

Veo (TFA using a solvent of 01.077 aqueous) 70.01 YA reversed-phase type C HPLC B solid 01" mg pure VY 0-+74 / to isolate an amorphous white. m, 2H), 1.67-1.76 (m, 2H), 1.77-1.84 (m, 1H), 1.92-1.99 (m, 1H), 2.22-2.08 (m, © 1H), 2.12-2.27 (m, 1H) , 2.22-2.27 (m, 1H), 2.6-2.67 (m, 1H, Pro-B'), 2.83-2.89 (m, 1H, Pro-p), 4.32 (dd, J= 12.1 & 3.5 Hz, 1H, Pro-3'), 4.41 (dd, J= 12.1 & 7.3 Hz, 1H), 4.56 (bd, J= 8 Hz, 1H,

Pro-8'), 4.62 (dd, J= 8.9 Hz, 1H, Pro-at), 5.4-5.46 (m, 1H), 5.55-5.61 (m, 1H), 5.73 (bs, 1H, Pro- 7) , 7.41 (d, J= 6.3 Hz, 1H), 7.64 (bs, 1H, Acca-NH), 7.80 (dd, J= 7.9 Hz, 1H), 8.03 (dd, J= 8 Hz, 1H), 8.07 ( d, J= 9.5 Hz, 1H), 8.16 (d, J= 7 Hz, 1H, AcNH), 8.36 (d, solution 8.3 Hz, 1H), 8.90 (d,J=6 .

Hz, 1H).

YYVY

Ve Example A (0) (Table 0008) Synthesis of compound 3 o d n b © : r 4a

NET + "ee r ve g 8 3 5 4 a A 5 . 3 f 0 i 0 : | 5 Zz 7 5 0 g 1 1 r ° 1 5 y r 1 1 ] 3

IVY

J pL 0 7# \ nb m 0 Ty 1 i

Lue oo yl J5

TY

58 q

Pu, i 23

A

01

a. Invert a solution of 54.97 g (01 mmol) of L-glutamine buffered by 71 Boc and 7 (a/g (YE) mmol) of (Y) 100000200 diacetate equivalent ) in 10 mL of a mixture of EtOAC/CH,CN/H,O0 (at a ratio of 0:7:7) at 10 C for 1 hour and at 10 C for ? hours. Then the reaction mixture was diluted with 01 mL of cold) and EtOAc 011,027 solvents were removed in vacuo and the remaining aqueous mixture extracted three times — diethyl ether (50 mL each time) and 56 mL of 206 to remove most of the Impurities. Then the aqueous layer (Alf) containing an intermediate compound of (amine) was concentrated to dryness and the residue was re-dissolved in 71 ml of :110.00 (at a ratio of 101); it was cooled to 0°C in an ice pleaa and a solution was slowly added of “ml (4 mmol) of benzyl chloroformate (equiv) in 0 ml of dioxane slowly over 0 about 0 minutes. The reaction mixture was stirred at 0°C for 1 hour die room temperature for 2 h. Then the mixture was diluted with 90 mL of water; extracted three times with cold diethyl ether (at about 00 mL each time; pang with HCl ( concentration (molecular) to a pH ranging from ? to V times EtOAc was extracted at an amount of 50 ml each time.The organic layers were dried after mixing them over MgSO, anhydrous and the ao was evaporated to dryness in a vacuum The raw material was purified by flash column chromatography, using a mixture of EtOAc/hexane/AcOH (with a ratio of (+,):Y,8:Y), yielding 0.744 g of compound 77b (the production rate was

Total (rey B. Anib YOu mg LEY mmol) of compound YT dipeptide intermediate Jo 171 mg (+83 mmol, 83 mmol) of compound QV VY ) © eq) and YAO mg (+089 mmol) of HATU (V,Y) eq) in 1 mL of CHCl added 1.794 mL (1.17 mmol) from DIPEA (; equivalents). The reaction mixture was stirred at room temperature for 164 hours, then the CHCl was evaporated in vacuum, the raw material was re-dissolved in 'EtOAc', and the EtOAc solution was washed with NaHCO; aqueous (concentration 70) and brine; and dried over MgSO, anhydrous

0

steamed to dryness. A mg FPA of SVT was produced after purifying the raw material by flash column chromatography; Using a mixture of EtOAc/hexane (at a ratio of V1) as the solvency material (the rate of

Production 478/). c. A solution of 75 mg (1.794 mmol) of compound 1Dv©mL of THF was cooled to 0 °C 0°C and a solution of VY mmol of ABH was added; 6.1 mL of dimethyl sulfide (01 mole solution; equivalents). The reaction mixture ld was brought to room temperature and stirred for 1 hour. It was then cooled again to 0° C before adding an aqueous solution of 0.8 mL ( 1.57 mM (Usa of NaOH (molecular concentration 0,¥; 0 equivalents) slowly over a time period of 11 minutes; then slowly (within about 1 min) a 0 aqueous solution of 1.8 mL (9, mmol) of AA)HO, molecular; 17.5 eq.) The reaction mixture was left to warm to room temperature and stirred for 1 hour. After this time period; aan The reaction was mixed to an approximate dagen degree; to hydrate the excess amount of (BH; then an aqueous NaHCO; was added to adjust the pH between about 4 and 10; 11177 was removed in a vacuum and the raw material was distributed between H, O and (EtOAc) and the aqueous layer B (EtOAc) was re-extracted and the organic layers were washed after vo mixing with brine; dried over SLY MgSO, and evaporated to dryness in a vacuum. The raw material was purified by flash vertical chromatography; using a mixture of EtOAc/hexane/NH,OH (at a ratio of 4:7:,) as the fluidizer; It resulted in 7 mg of the pure compound 7H (average

Production (rev d.) Add 140 mg (77© mmol) of Dess-Martin periodiante (at a ratio of 749#0 eq)© to a solution of compound 71 H in A ml of CHCl and the reaction mixture was stirred at room temperature for an hour and a half The reaction mixture was quenched by adding ¥ ml of aqueous Nas$. vo min. of jal, the reaction ore was extracted with 2086 and the organic layer was washed with aqueous NaHCO

a

0 anhydrous and evaporated in vacuo yielding 188 mg of MgSO (concentration 7.0)” and brine and dried over (78W) which was used in the next step without further purification. aldehyde mmol) of compound (17O) and 78 μL Mn +, YY) mg 188 AH. Stir a solution of room temperature ethanol in ¾ mL of PA(OH), 11.001 mg of Yo and

Hy, an hour. And after this period; More gas 11 at atmospheric pressure was added for a period of hydrogen | 0 The stirring flask was continued into the CH;COH flask with 1 μl of Y0¢ mg from (70) 017 and 1868 and an additional hour. then filter the mixture and evaporate the solvent to dryness; The bulk cyclic product YE of the crude was purified using chromatography by vertical chromatography with a mixture of macrocyclic product (production rate (OY) of the compound pile £A (7:10 ratio):) resulting in CHClL. /MeOH/AcOH (Ye about 0 μl OY mmol) of compound +0 FY) mg YY mixture of STL (07+ mmol) of AV equivalents) (©) DIPEA (mmol) of +,100) h. Then, 11 at room temperature for CHCL, eq.) in © ml of acetic anhydride (11 mg) and (V:Y:Y) (with a ratio of 0.011 THF/MeOH/H) was removed. 01 in a vacuum and add © ml of a mixture of H at TA equivalents) and let the hydrolysis reaction proceed for 0 (LIOH2H,0 mmol) of TY) 0 reaction mixture (to a pH of pal M Then, at 20 room temperature and for 2 hours at 1 mg of the final compound 508 (reducing 1 inverse-phase HPLC yield of about ;) and purified at a production rate of about 7 for the last two steps). DMSO, 400 MHz) of 508 (mixture of rotamers confirmed by COSY, TOCSY and ROESY NMR data): 1.18 (s, 9H), 1.09-1.85 (overlapping m, 11H), 1.95 (s, 3H), 2.3 (m, ¥.1H), 2.63 (m, 1H), 3.18-4.14 (overlapping m, 6H), 3.96 (s, 3H), 4.44 (m, 1H), 4.62 & 4.69 (2d,

J=11.8 Hz, 1H, rotamers), 5.82 (bs, 1H), 7.2 (m, 2H), 7.53 (bs, 1H), 7.67 (bs, 4H), 8.19 (bs, 3H), 8.61 (s, 1H) ).

Ya

YY Example { Yv ) Saturated Macrocyclic Intermediate (Ala) Synthesis of an intermediate compound

Oh

09

Da 8 and BtOAc a BoCHN N, o alumine mounted on Rh (Hp (Ve) (concentration 7 up) of on 9 0 9 0 atmosphere EA, He NP

Loo Ths fry QTY ow | Equivalent %A (LIOH.H,0 0 om oH 0 0 8 — Da 8 " ِ for enumeration 1 ivy unsaturated ag ad all mmol) of the cyclic intermediate VT) g Yo A. dissolved 0 mg 001 ml of 2104 and added 0 bp in YY saturated macrocyclic intermediate (concentration 70) and stirred in an atmosphere of Rh alumina gas by weight) of 71 ) at atmospheric pressure and at room temperature for an hour and a half. After this paragraph; hydrogen

YAY

HPLC analysis confirmed the complete transformation of the feedstock into two products; Desired product IVY and by-product cyclopropane formed from ring opening (YY in compound Ga by total mass) defined as 7A) and filter the reaction mixture and concentrate yielding VEY g of a light green solid. The solid was evaporated twice jointly with EtOAc to remove all EtOAc (Cus 0:5 presence leads to the incompatibility in the next step). It was proved that the separation of compound 71?a from compound 77b by chromatography is very difficult. Hence, an alternative method was devised based on the relative rates of hydrolysis of the respective moieties of B.methyl ester. 7.47 g of the crude mixture of compounds 7?a and 71b were dissolved in 01 mL of a mixture of ‘THF (dui) MeOH 1:0); an aqueous solution of YE was added 1 g of LIOHH 0 in mL of 0 water (78 equiv) and the reaction mixture was stirred at room temperature for 11 hours (complete hydrolysis of side product 1b to its corresponding acid YV cb (proved HPLC The reaction mixture was concentrated in a vacuum to remove most of the THF and MeOH and distributed between 100 ml of water and 60 ml of EtOAc and the organic layer was washed three times with NaOH (0,+N) of 001 mL each time; and 100 ml of brine water; and twice — AW citric acid (in ratio)/01 by 100 ml in ve each time.” and 1,000 ml of brine; and dried over MgSO, anhydrous; and filtered and concentrated to dryness. 7,748 g of the desired product 7?a was produced with a high purity (>740b HPLC) light green foam form (production rate was JS 797 for the two steps). 'H NMR (400 MHz, CDCl): 6 1.1- 1.38 (m, 13H), 1.42 (s, 9H), 1.51-1.57 (m, 1H), 1.63- 1.67 (dd, J= 8.0 & 5.1 Hz, 1H), 1.81-1.87 (m, 1H), 1.92 -1.99 (m, 1H), 2.02-2.08 (m, 1H), 2.62 (d,J= 14 Hz, 1H), 3.4 (d, J=8.3 Hz, 1H), 3.65 (s, 3H), 4.01 ( dd, J= 10.8 & 4.1 Hz, 1H), 4.42-4.48 0 (m, 1H), 4.51-4.55 (m, 1H), 4.87 (d, J= 8.6 Hz, 1H), 5.14 (d, at ‎ 8.6 Hz, 1H), 7.97 (br.s, 1H).

Example YA Synthesis of Compound No. VEY (Table (V OH Boer rn 8 3 OMs + 5 0 0 — A "ab Y)D1AD equivalent) ¥ ; | N, Y)Ph,; P equivalent), LIOH(Y OH) is s A CH! N. | / (T° A 2 BocHN N A 7 3 o KK; OH 65 —— the compound VEY ° The quinoline derivative is bound to the previously formed macrocyclic compound 11b by means of the Mitsunobu reaction. 0 mg (+, 30 mmol) of 5A quinoline derivative in (THF) then add 0,£ pile of megacyclic 1 no

1 (YY macrocycle equivalent) and 55.8 mg of (Y) PPh; equivalent). The resulting mixture was cooled to zero degrees Celsius. Then 7.4 µL of DIAD (¥ equivalent) was added dropwise. The solution was stirred for one hour at 0°C and then stirred overnight at room temperature. Then the mixture was diluted with 0 ml V of BtOAC and washed with 11 ml of a saturated solution of NaHCO; Yellow oil.The product was purified by flash chromatography on silica gel (using M8086 by 100).The product still contained DIAD by-products after purification.The resulting product contained 700 by weight of the desired product.It was then found that the production rate SAY was dissolved 7 lb mg (501+ mmol) of an intermediate compound of 1 (ester mL of a mixture of THEMeOH/H,0 | 0 (with a ratio (V::Y):Y) and 01 was added mg EA (+mmol) of LIOHH,0 and stir the solution at room temperature. After a time period of 116 hours; By HPLC reaction mixture analysis, hydrolysis has been completed. The organic solvents were removed in vacuo and the remaining raw material was dissolved in (CH5),SO by reversed-phase HPLC type VA to produce the pure inhibitor VEY '" H NMR (400 MHz, DMSO-d): 6 ( ppm) 8.67 (s, 1H), 8.29-8.14 (m, 2H), 8.08-7.97 (m, m0 1H), 7.91-7.78 (m, 1H), 7.74 (s, 1H), 7.31-7.2 ( m, 1H), 7.10 (d, J=5.7 Hz, 1H), 5.82-5.71 (m, 1H), (m, 1H), 5.32-5.23 (m, 1H), 4.74-4.64 (m, 1H ), 4.55-4.47 (m, 1H), 4.23-4.06 (m, 1H), 5.58-5.47 (m, 1H), 3.97 (s, 3H), 3.92-3.85 (m, 1H), 2.7-2.55 (m, 2H), 2.53-2.36 (m, 2H), 2.2-2.09 4.04-3.94 (m, 1H), 1.8-1.62 (m, 2H), 1.56-1.43 (m, 2H), 1.42-1.29 (m, 6H), 1.27 (d,J=3.2 Hz, 3H), 1.25 (d, Hz, 3H), 1.12 (s, 9H). 2.9 -1 MS: 763.1 (M+1), 761.1 (M-1).Y

Example Ya Synthesis of compound number Yoo (Table (Y Cr, COCH; i ] 3 (v2) 1 tydm a 5 8 N zo H 8 ~ 7" boom

Veo compounds A solution of 1.6 mL (Yo, + mmol) of 050 in t-butanol (at a ratio of 70 w/v) was added to a solution of 7 mg) °C. mmol) of the #1 macrocyclic compound in a mixture of 0 V.0 mL of V.0 5 t-butanol mL water at zero degree li’ (5 gall) the mixture at room temperature for one hour.The mixture was diluted with 71 ml of BIOAC and the organic solution washed twice with NaHCO (concentration of 70) at an amount of 0 ml each time and twice with brine water (at an amount of 10 ml each) Bae was dried and evaporated to dryness.The crude compound was dissolved in a mixture of ¥ ml V.0/THF ml from 1.8/146011 ml V.

a

1 hour. Mixture 11 was acidified for 0.0174 LIOHH of (Use mM 0174) mg VY and stirred in the presence of H,0

HPLC to a pH of ; using 1101 ice-cooled (10+ gauge); Pure B of 46 (reversed-phase TFA of type C81) was evaporated using a solvent graded of water (7000) of high purity as a solid of 705 adjacent to an aqueous diol (70.05 of 178). and isolate CHCN, white, amorphous. ), 1.78- 1.88 (m, 1H), ~ 2.2-2.5 (2m, 2H), 3.78-3.82 (m, 1H), 3.86-3.90 (m, 1H), 4.39 (t, J= 8.9 Hz, 1H), 4.61 (d, J= 11.4 Hz, 1H), 5.60 (bs, 1H, Pro-y), 7.03 (d, at 6 Hz, 1H), 7.4 (bs, 1H), 7.58-7.62 ( m, 1H), 7.87-7.91 (m, 1H), 8.00 (d, J= 8.3 Hz, 1H), 8.24 (d,J=8.6 Hz, 1H), 8.6 (s, 1H), 8.99 (bs, 1H) ). 0

EMS (negative ionization mode): m/z 625 (M-H).

VAY veo Example (Y) (Table YVA and 7116) Synthesis of the two compounds

OMe OMe

BY

N i © J b * at 7 › Cyt s LD

N N g © o 0 ir. A27+. or a

ome. OMe or, OY,

JS = 1l 5 7114

OMe OMe co oe 7 Egle 718 The boat © Ki °

YYVY

a a. Stir a solution of 81.10g (+5 mmol) of 11-keto-nonenoate ester d in a mixture of ©mL of MeOH and 7 mL of water at room temperature in the presence of 50 mg (V,Y) mM (Use of LIOHH,0 for 1 hour. The solution was acidified to pH > using ice-cooled 1101 (0,+N) and most of the cull MeOH concentrate was evaporated in 1 mL © 8086 and the solution washed with 01 ml of 110 icy coolant (+0, standard); and 10 ml of brine water; And dried and evaporated. The ore was re-dissolved in 10 mL of CHCl, and reacted with TTY g (+571 mmol) of the 0 012 fraction a in the presence of “!7 mg VY (+ mmol) of HATU and 10 µL (7.4 mmol) of DIPEA over a time slot of 116 h at room temperature.The reaction mixture was sorbed on silica/hammer using a mixture of EtOAc/hexane. (to a ratio of 0:1) as the solvent to isolate 0.17 g of the pure compound Fe (yield rate AY and purity > 780 as specified — HPLC '" H NMR (CDCl, 400 MHz) : § 1.41 (s, 9H), 1.45-1.54 (m, 1H), 1.58-1.62 (m, 1H), 1.73- (m, 1H), 1.86-1.91 (m, 1H), 2.16 (dd, J= 17.8 & 8.6 Hz, 1H), 2.26-2.43 (2m, 2H), 2.46-2.58 1.77 (m, 2H), 2.64-2.81 (m, 1H), 2.85-2.92 & 2.95-3.03 (2m, 1H), rotamers in 1:3 ratio), 3.67 (s, 3H), Vo (s, 3H), 4.10-4.18 (m, 1H), 4.20-4.30 (m, 1H), -4.40 4.55 (m, 1H), 4.8-4.88 (m, 1H), 4.92- 3.95 (m, 2H), 5.14 (dd, J= 10.2 & 1.6 Hz, 1H), 5.24-5.38 (m, 4H), B . A ring of 0.77 g (0.89 mmol) of CF diene was formed in the presence of 1217© mg (1 mmol Y) of the catalyst bis (tricyclohexylphosphine) benzylidene ruthenium IV dichloride in CH,CL, (distilled from die J) jay Cal, gas — Ye sad argon min) over a two-hour period at reflux temperature. IVE produced a gram of the compound in the form of a mixture of the two stereoisomers (1 "C and OF) with a ratio (Vi) after achromatography.

A snapshot in a column on a silica gel using a mixture of EtOAc/hexane (by 7) (production rate was 5 77). NMR of mixture 30c & 30d (CDCl;, 400 MHz): § 1.44 (5, 4H). ) & 1.37 (s, 4H), 1.6 (m, 11 2H), 1.83 (m, 0.5H), 2.01 (m, 1H), 2.09 (m, 1H), 2.42 (m, SH), 2.73 ( m, 2H), 3.26 (m, 0.5H), 3.69 m (s, 1.5H), 3.76 (s, 1.5H), 3.96 (s, 3H), 4.10 (m, 1H), 4.24 (m, 0.5H), 4.10 (m, 0.5H), 4.58 (m, 1H), (m, 1H), 4.89 (m, 0.5H), 4.97 (m, 0.5H), 5.30 (m, 0.5H) , 5.44 (m, 2H), 5.64 (m, 1H), 7.1-7.0 4.73 (m, 3H), 7.47 (m, 4H), 8.08-7.98 (m, 3H). 7 mg TY (0 mmol) of JS of methyl ester 1"gu 1d in a mixture of 1 mL of THF 0 mL of 148011/5, mL of 11:0 using 11 mg 0 (714, mmol) of 0.LIOHH over a time period of VT 1 hour at room temperature. After this period oan the reaction mixture to a pH of -9 ; C81 reverse-phase HPLC column using a solvent gradient of water (70.0 of TFA) to 0 of 0 lm (700) of pile of TFA of each of the two desired compounds (716). and 708 of the mixture of the two compounds with a high purity (64 as determined by HPLC) (the production rate was 10) (Ye (YY ES ll 'H NMR (DMSO, 400 MHz)): § 1.15 ( s, 9H), 1.48-1.54 (m, 2H), 1.65-1.74 (m, 1H), 1.77- (m, 1H), 2.12-2.25 (m, 4H), 2.27-2.34 (m, 1H), 2.61-2.68 (m, 1H), 2.87 (bt, J= 11.5 Hz, 1H), 1.85 (dd, J=9.2 & 1.5 Hz, 1H, Pro-8), 3.97 (s, 3H, -OCHj), 4.14-4.20 (m, 1H), 4.52 (t, J= 7.8 Hz, 3.92 1H, Pro-0), 4.66 (d, J= 11.8 Hz, 1H, Pro-5), 5.45 ( t, J= 9.9 Hz, 1H), 5.51-5.58 (m, 1H), 5.82 (bs, + IH, Pro-y), 7.09 (d, J= 6Hz, 1H, BocNH), 7.26 ( bs, 1H), 7.53 (s, 1H), 7.67 (bs, 3H), 8.16 (d, I= 2Hz, 1H), 8.18 (s, 1H), 8.83 (s, 1H, ACCA-NH). YiVY

VY. (YA compound'H NMR (DMSO, 400 MHz): § 1.06-1.10 (m, 1H), 1.18 (s, 9H), 1.52-1.55 (mm, 1H), 1.62- 1.8 (m, 1H), 2.1-2.68 (overlapping, 9H), 3.90 (bd, J= 8.3 Hz, 1H), 3.96 (s, 3H, OCH,), 4.20-4.27 (m, 1H), 4.58-4.63 (m, 1H , Pro-8), 4.66 (dd, J= 8.3 Hz, 1H, Pro-a), 4.88 (dd, J= 10.2 Hz, 1H), 5.18-5.26 (m, 1H), 5.73-5.79 (m, 1H , Pro-y), 7.01 (d, J= 6.4 Hz, 1H), 7.23 (bs, 1H), 7.50 (bs, 1H), 7.66° (bs, 3H), 8.20 (bs, 2H), 8.53 (s , 1H).

IN oN

SAL 11 5 SLL rE ~~ - 1 _° AN © Pa Ns © Rar (J 2 2 3 o E A o 0 9 . 0 +

SLI TTY LADY YT SLA H N SAH 0 H H “ny 6

Yq Ag | Compound 1 only

011 FY CHCl removed ml of “0 a in 1 diene mmol) from +, TF) A. Dissolve 7 44 mg mg (0.0 mmol) of catalyst AY and dissolve Yo for argon gas from the solution by adding in ?ml of .11.01 anhydrous bis (tricyclohexylphosphine) benzylidene ruthenium IV dichloride and the reaction mixture was heated with reflux for 2 hours in degassed diene and the product was added to the solution mg of WY and the solution was concentrated and purified by flash chromatography until nitrogen (.)/7971 yielded compound 1b as a brown solid (yield 'H NMR (CDCl, 400 MHz): § 1.22 -1.44 (m, 10H), 1.42 (s, 9H), 1.66-1.74 (m, 1H), 1.87- 1.97 (m, 2H), 2.13-2.28 (m, 3H), 2.32-2.39 (m, 1H) , 3.08-3.16 (m, 1H), 3.41 (s, 3H), 4.07-4.22 (m, 3H), 4.28-4.34 (m, 1H), 4.58-4.64 (m, 1H), 4.95-4.99 (m, 1H), 5.22-5.29 (m, 2H), 5.38-5.43 (m, 1H), 5.48-5.56 (m, 1H), 7.0-7.12 (m, 3H), 7.43-7.55 (m, 4H), -7.97 8.11 (m, 3H).)

ES(+)MS: m/z 727.4 (M+H)". ml of 1101 N with a concentration of 1 mmol of compound 1b in a solution of 1.117 B. Stirred, then CHCl for 1 min. and concentrate to dryness.The solid product was dissolved in ¥ ml of 71 dioxane in mmol) of 751 mg YO (4 BLN mmol) of 0 μL ONO) AY was added. hr; concentrate Yo sequentially. After stirring at room temperature for t-butylisocyanate Vo, the mixture was reduced to dryness and the resulting crude compound was used 17 g in the final hydrolysis step without further purification. (1 L and THF mmol) of compound 1c in a mixture of ?ml 1 V) mg of Ao C. dissolved and stirred the solution for 0 LIOHH, 0 mg (+477 mmol) of YA then added 11.0 Js) and MeOH acetic acid was added to 1 ml of 1 hour cells at room temperature, after the passage of the time period 0 Y. mg of pure compound 4 after purifying the product YO and 1117. MeOH was isolated and the solution was concentrated to remove (7%) (The production rate was about 1 inverted-phase type C HPLC raw material by a

'H NMR (DMSO, 400 MHz): 5 1.04 (s, 9H), 1.15-1.24 (m, 2H), 1.30-1.40 (m, 5H), 1.44- 1.51 (m, 2H), 1.54-1.68 (m , 1H), 1.75-1.88 (m, 1H), 2.18 (dd, J= 17.2, 8.5 Hz, 1H), 2.32-2.45 (m, 1H, Pro-B), 2.54-2.62 (m, 1H), 2.65 -2.68 (m, 1H, Pro-g), 3.91 (dd, J= 11.1 & 3.5Hz, 1H, Pro-5), 3.96 (s, 3H, -OCH,), 4.17-4.23 (m, 1H), 4.47 (dd, J=8.6 Hz, 1H, Pro-e), 4.67 (bd, J=7.9 Hz, 1H,

Pro-8), 5.30 (dd, J=9.5 Hz, 1H), 5.52 (bdd, J= 19 & 8.3 Hz, 1H), 5.68 (s, 1H), 5.78 (bs, 1H, Pro- °v), 5.94 (bs, 1H), 7.21 (bs, 1H), 7.51 (bs, 1H), 7.66 (bs, 4H), 8.19 (s, 2H), 8.4 (d, J= 7 Hz, 1H), 8.61 (s , 1H, ACCA-NH).

ES(+)MS: m/z 698.3 (M+H)".

PY Example for Lm “oor 0 o 0 a o 9 0 8 y 0 N 8 4 p.

a a 9 ? 9 0 NN i 8 8 “oH 7 “om 7 BocHNU L. o rnd 6 7 compound 04 - Teves, a. Dissolve 84 mg (+, mmol) of diene 7?a in 1 ml of anhydrous O011:0 and degasify the solution over a period of time of Yo min with a flow of .argon and solute or Y 4 mg 9 67 mmol) of M bis (tricyclohexylphosphine) benzylidene ruthenium IV dichloride catalyst in 1 mL of degassed CHCl was then transferred to the reaction flask using a cannula. The reaction mixture was stirred for 2 hours at reflux temperature. Then the solvent was removed under vacuum and the reaction mixture was purified by flash column chromatography on silica gel using a mixture of 1% hexane g ethyl acetate as the liquefaction agent; So FY yielded mg of cyclocyclic 1b in the form of yellow oil (production rate of 741). That's why. Dissolve vy mg (0°, mmol) of an intermediate compound of VY ester in ¥ mL of a mixture of THF/MeOH/H,0 in a ratio of 1:7:A then add (mg A) ,+ mmol) of LIOHH,0 and stir the solution at room temperature. After a time period of V1 hour, analysis of the reaction mixture by HPLC indicated that the hydrolysis was not complete. Thus, an additional amount of ; mg (+09 mmol) of LIOHHO and the solution was stirred at room temperature for a total time of v h. jm 2S aly solution with a small amount of acetic acid, organic solvents were removed in vacuum, and the remaining raw material was purified by reverse-phase HPLC solution

184 . 504 until pure inhibitor yield 1 C "HNMR (DMSO, 400 MHz): 5 1.21 (d, J= 6.0 Hz, 3H, Me), 1.36 (s, OH, Boc), -1,10 1.40 (3m, 3H), 1.66 (m, 1H), 1.8 (m, 1H), 2.10 (m, 2H), 2.57 (m, 2H), 3.90 (m, 4H), 4.47 (bd, J= 12.7

Hz, 1H), 4.58 (bd, J=7.3, 1H), 4.66 (dd, J=8 Hz, 1H), 5.57 (m, 1H), 5.66 (m, 1H), 5.83 (bs, 1H), 6.18 (bd, J= 6.9 He, 1H), 7.25 (bd, J=7.3 Hz, 1H), 7.56 (bs, 1H), 7.70 (m, 4H), 8.22 (bd, J=2.0 +

Hz, 2H), 8.29 (bs, J= 9.2 Hz, 1H). pale ¥ ethanol (mmol) of the inhibitor ¢ £4 was added in ¥ ml 1071) mg Vo C. It was dissolved at a temperature of hydrogen held on ©. The mixture was stirred in an atmosphere of 0 concentration Pd of 18 hours inverted-phase HPLC, and after filtration, the mixture was purified in a VT chamber for a period of 777 μg white solid product (production rate was GEV) 01 mg of inhibitor until we get 0

HNMR (DMSO, 400 MHz): 1.04 (m, 1H), 1.17 (d, J= 6 Hz, 3H), 5 (s, 9H), 1.25- 1-75 (m, 12H), -2.32 2.45 (m, 1H), 3.40-3.50 (m, 2H), 3.74-3.83 (m, 1H), 3.85-3.95 (m, 1H), 3.97 (5, 3H), 4.27-4.36 (dd, J=21.1 & 8.6 Hz, 1H), 4.54 (dd, J= 7.95, 7.95 Hz, 1H), 5.64 (d, J= 8.3 Hz, 1H). 5.82 (brs, 1H), 7.27-7.33 (m, 1H), 7.53-7.57 (bs, 1H), 7.6-7.74 (m, 4H), 8.13-8.27 (m, 3H), 8.3-8.35 (brs, 1H ). Yo

1010

FY Example | 3 : 3 = i Lh 0 1 >

Tz 3 2 or 3 dam Lo “rz © 5 1 3 oF = [=> 3 * 1 0 . 3 g = z 3 3 0 9 3 1 2 2 T x 2 5 © 1 { 3 | Ham 3 1

S x az! y . 8 0 5 3 4 3 =z = ng, & A 2 J g 13 5 > z <

CT 8 oo = 8 © and d 3 Jot 0 z adsorbent mL of :013:01 and degassing of 0 mmol of compound ??a in 1.96 a . dissolve about

YYVY

11. Carefully shake the solution before adding a sample of 17 mg YA (0 mmol) of Hoveyda's catalyst and then stir the solution with heating at reflux temperature for ½ hours. The reaction mixture was concentrated and purified by flash column chromatography; using a solvent of CHCL/EIOAC in a scaling ratio of 7: to 7:7 to yield 194 mg of GFF (production rate (IVY) 0 pala 0 μl + ,Y1) mM mol) of DIAD (© equivalent) slowly to a solution of 70 mg (0.147 mmol) of ry and 17 mg YAS (mmol) of 2-ethoxy-4-hydroxy-7-methoxy -quinoline VAT, mg (91 mmol) of PhP (©equivalent) in 0 mL of anhydrous THF at zero gestational temperature within twenty minutes. The reaction mixture, Bad, was left to room temperature and stirred at this temperature for two and a half hours. And then - after that” AD 2117 in a vacuum and pure the crude product by flash column chromatography using a solvent of hexane/EtOAC at a ratio of 17:7 to 1. and separating VY mg from the pure compound “JB” (the rate of Yield (vy c. Mixing VY 1004 mg(mmol) of compound (—a¥'T) with ¥ mL of CHCl, *mL of 110 concentration; mole in dioxane The mixture was left to stir at room temperature for an hour-and-a-half to separate the protective Boc group and obtain the !110 salt of the intermediate compound IY and evaporate the crude reaction mixture to dryness in a vacuum; vacuum dried to ensure removal of all amount of 1101 and use in the next step without Purification d. in 0 7 mL of THE and the mixture was stirred at room temperature for 2 h to form a reactant of cyclopentyl chloroformate after which dial time approximately half of the solvent was removed by evaporation into a vacuum; the remaining light yellow solution was diluted by the addition of © mL of CHCL reconcentrated to half its original volume; to ensure all excess phosgene is removed. And dilute the solution of substance A

The cyclopentyl chloroformate prepared above was further reacted with 01 mL of ‘THF Sa’ at 0 °C and added to solid compound ‘d (£ 0 mmol) at 0 °C. Vo μl (078; mmol) of BUN (0.7 equiv.) was added to the reaction mixture and stirring was continued at 0° for an hour and a half. Solvents were removed in +vacuum and the raw material was purified by flash column chromatography; using a mixture of 110801608 at a ratio of 1:1 as the evaluating agent until VO mg of the compound (77e) was yielded at a quantitative yield of approx. H. Hydrolysis of a methyl ester was carried out by reacting mg Vo (1 mmol) of compound E with To mg AE (mmol) of 1:0111:0 (A equivalent) in 7.5 ml of a mixture of solvents THF/MeOH/H,0 in a ratio of 1:7:7 at 0C for 2½ hours. and after completion - hydrolysis; mt A the mixture was reduced to a pH of £0.0 and the solvents were evaporated to dryness in a vacuum. The crude product was purified by a YA “C18 reversed phase preparative liquid chromatography” using a solvent of H,0 and CH;CN with a concentration ranging from zero to 54 (in addition to 758 at a concentration of is 50,05/); So £0 mg of synth 4 yielded as a white amorphous solid (yield rate 7710) ‎of the 1127 salt of #824 (DMSO, 400 MHz): § 0.88 (d, J= 6.7 Hz, 3H), -0.95 1.70 Vo 1111/1 (overlapping resonances, 17Hz), 1.37 (t, J= 7 Hz, 3H), 2.00-2.10 (m, 1H), 2.10-2.33 (m, 3H), (m, 1H ), 3.8-3.85 (m, 1H), 3.85 (5, 3H), 4.02-4.08 (m, 1H), 4.42 (q, J=7 Hz, 2H), 4.35-2.38-2.44 (m, 1H ), 4.50 (d, J= 10.8 Hz, 1H), 4.63 (bs, 1H), 5.28 (dd, J= 9.5 Hz, 1H), 5.38 (bs, 1H), 4.44 (m, 1H), 6.37 (s, 1H), 6.87 (dd, J=8.9 & 2.2 Hz, 1H), 7.07 (d, J= 2.2 Hz, 1H), 7.28 (d, 5.42-5.49 1H), 7.90 (d, J= 8.9 Hz, 1H), 8.57 (s, 1H).2,2L 1-7

Example ve Synthesis of the compound AMY (Table A) 9 Loon OMe OH 0 o 8 ° 0 for # A N © "SN OMe men i free NHOH 0 NH 0 —_— l l-0 7 i 0 PhyP; DEAD - 0 0 YY iy ¢ no yva 0 0 N, (oe ° and “OE i 0 NH oo” ove 70 1 0 (adjacent) 0 . N, N,

J~ Jd 8

YY Y 0 6 0 NH 0 NH

LF - LE

Lave ° art ° —A oy oo of oe #

ORD

I 1 I] 3 ~~ 7 ~~ ave Sr©

YYVY

5 00 1 oA a | ow MN Dans Free Free 0 p. 1 oF

YY OY es ve ve © ° boat 817 a. ©0117 mL (04.7 mmol) of Y (DIAD equivalent) was added drop by drop during Vo min to a solution of 03.06 g (77.7 mmol) of a solution of the cyclic intermediate Bulk 1 (macrocyclic intermediate equivalent) VEYA mg (0.8 mmol) of PhaP (equivalent) ° and ,11 g YA (mmol) of 2-carboxymethoxy-4-hydroxy-7- methoxyquinoline (Gg) (1880), 0 0) for IPOs 0/0484 and (0<1/04088) in £00 mL THF at 0°C. Then the ice bath was removed and the reaction mixture was stirred at room temperature for 2 hours. And after the completion of the transformation of the raw material into products; The solvent was evaporated under vacuum and the remaining mixture was dried with EtOAC, washed with NaHCO; saturated twice and with brine once, and the organic layer was dried over MgSO, anhydrous, filtered and evaporated to dryness. Pure compound 4"a was obtained after a PLC procedure; the column was firstly mixed with hexane and 2100 (at a ratio of 2:0) and then mixed with a mixture of EtOAc CHCl (at a ratio of 0:90) to remove 1:70 and the products of secondary DIAD and monitor the liberation of impurities by J pal TLC thin layer chromatography of the desired product on a column containing a mixture of EtOAc CHCl (700:760 ratio). times before separating the compound TE with high purity in the form of a white solid product (the total production rate was (TA) and resulted in 17.8 g of the desired compound with a purity of

171

HPLC was 74.0 by dioxane to a solution of 1,917 g dioxane; Standard in HCL of 1 ml b. The reaction mixture was added and stirred at a temperature of 15 ml CHCl in Boe b (sal). The intermediate compound partially formed a thick gel during the reaction. Ala was added to the room for one hour [in An additional ml of .611:01. After the removal of the protection was completed, the solvents were evaporated to dryness until a product of 78 0 01 was produced with a concentration of about MeOH, a yellow solid and a paste-like substance. The mixture was re-dissolved in 0,011,A It was re-evaporated to dryness in a vacuum until compound B was produced in the form of a yellow solid product that was used in the next step without purification. Its concentration was 1.47 molecules in phosgene c. cyclopentanol μl (7 mmol) of TYE solution dropwise to a solution of toluene Ve mixed at room temperature for two hours to form CC B85 THF ml of VO in cial volume and after During that time period; 0 (adjacent) cyclopentyl chloroformate reactant was removed from the solvent almost by evaporation into vacuum; the remaining light yellow solution was diluted with excess phosgene JS and concentrated to half its original volume to ensure removal. CHCl Add © ml of the above preparation additionally cyclopentyl chloroformate and mix the reactant solution of 1s and cool the mixture to zero degree TE amine2HCI and add to THF salt ml of V0 using and adjust the pH to a value of approximately 4-8.5 gals celsius in a drop by drop bath and the reaction mixture was stirred at 0°C for one hour. BON by adding

NaHCO; washed with water once and with EtOAC b ge yall and after that period of time; anhydrous dilute; MgSO, twice saturated; And 11.0 twice and brine water once. The organic layer was dried over © 1,717 gm, filtered, and evaporated in a vacuum until a reddish-yellow foam was produced. The compound (*C) was produced in the form of a white foam after purification by flash vertical chromatography as a liquefaction agent (EtOAc) at 7700-7750 with a hexane concentration (using a 1st solvent).

The yield was (A) purity > JAY

Dr.. Y dissolve 1.1 g of dimethyl ester; c in 10 mL of a mixture of THE/MeOH/H,0 in ratio

It is 9:1 and an aqueous solution of VA was added to 1 mL of NaOH of concentration 1 N (equivalent).

The reaction mixture was stirred at room temperature for 1 hour before evaporating to an even dryness

0 Approximately 1.176 mmol of the OVE sodium salt was produced as a white solid. And use the compound

VE in the next step without purification.

h. Dissolve 7 mmol of crude sodium salt OVE in 117 ml of THF and add 8037

The mixture was cooled to 0°C in an ice bath. 77 µl (0,¥ mmol) was added.

of isobutyl chloroformate dropwise and stirred the mixture at 0°C for 1V min. 0 and after that time period; VO mL of diazomethane was added and stirring continued at zero °C

semen for Te min and then at room temperature for an additional del. Most of the solvent is evaporated

dehydration in vacuum and remaining mixture diluted with EtOAC and washed with NaHCO; saturated twice;

twice with water and once with brine over MgSO, anhydrous”. It filtered and evaporated to dryness

Until 1.7 g (about 1.17 mM) of ATE resulted in a light yellow Vo foam. An intermediate compound of diazoketone ;E was used in the next step without purification.

0.7 S09 g (1.17 mmol) of diazoketone 4 7E dissolved in 117 mL of THF to

zero degrees Celsius in an ice bath. 4 ml of an aqueous HBr 758 solution was added

Drop by drop and stir the reaction mixture at 0°C for 1 hour. Then dilute the mixture

b (EtOAc) and washed with NaHCO; saturated twice; with water twice and brine once; and ye dried the organic layer over MSO, anhydrous; filtered and evaporated to dryness until approx.

0.609 mmol is approximately a light yellow foam.

g. Add MIA mg (1.90 mmol) of thiourea (a solution of 000 mg

(779© mmol) of a 4” bromoketone solution in ©mL isopropanol and set the reaction mixture

a

In preheated ziti ales at L©am, it was left to stir for one hour. Then the isopropanol was removed in a vacuum and the product was dissolved in 100 ml of (EtOAc) and the solution was washed with NaHCO; OYY mg of the crude product (; Crude methyl ester tA (FE) mL of solution 00 in a ratio of 1:1:7 was reduced to a soapy consistency using AQ 7.06 mg (mmol) of .LIOHH;0 and the hydrolysis reaction was carried out over a period A time of 11-19 hours at room temperature. Then the solvents were removed in a vacuum and the crude product was purified by C81-type inverted ye-phase HPLC, using a solvent of 011.00 by 11.0 with a concentration of 01 to 0 until a mg of YE was obtained. BAY Y HCV protease inhibitor HCV Product image yellow solid (total conversion rate of the oT intermediate to the AVY inhibitor was ./718 'H NMR (400 MHz, DMSO-dy): & 8.63 ( s, 1H), 8.26-8.15 (m, 2H), 7.79 (bs, 1H), 7.72 (bs, 1H), 7.5 (bs, 2H), 7.33-7.25 (m, 2H), 5.77 (bs, 1H) , 5.52 (dd, J= 8.3 Hz, 1H), 5.27 (dd, J=9.2

Hz, 1H), 4.64 (d, J= 10.8 Hz, 1H), 4.5 (dd, J= 8.3 Hz, 1H), 4.39-4.31 (m, 1H), 4.08-3.99 (m, 2H), m 3.94 (s, 3H), 3.87 (d, J= 9.5 Hz, 2H), 2.65-2.53 (m, 2H), 2.46-2.36 (m, 2H), 2.2-2.12 (dd, 1-6

Hz, 1H), 1.8-1.64 (m, 2H), 1.63-1.06 (m, 14H).

MS; es”: 733.2 (M+H)”, es: 731.2 (M-H)”.

YAVY

Example ire The compound (A) 11 (see Table A) was prepared using the same procedure described in the example; but by reacting bromoketone ¢ or with commercially available N-methylthiourea. Ne 7 H o dr A 0 Q N N § m" 0 | 5 * 2 0 0 om 20 _— 'H NMR (400 MHz, DMSO-dq): 5 8.63 (s , 1H), 8.20 (s, 1H), 8.18 (s, 1H), 8.12-7.93 (m, ¢ Hz, 1H), 5.28 8.1 per 1H), 7.88-7.69 (m, 2H), 7.32 -7.24 (m, 2H), 5.82-5.75 (m, 1H), 5.52 (dd, (dd, J= 9.9 Hz, 1H), 4.67-4.61 (m, 1H), 4.51 (dd, I= 8.8 Hz, 1H), 4.44-4.37 (m, 1H), 4.08-4.00 Hz, 3H), 2.65-2.37 (m, 3H), 2.16 (m, 1H), 4.1 each (m, 1H), 3.96 (s, 3H), 3.89 (m, 1H), 3.04 (d, (m, 2H), 1.63-1.11 (m, 17H). 1.77-1.65 MS; es” 747.2 (M+H) ", es': 745.3 (M-H)" Ve

and example; b Compound 801 (see Table A) was prepared using the same procedure described in example TE but by reacting with bromoketone; and with commercially available N-ethylthiourea.

A < free ° : 1 OD = : 8 / o © 7 é OH Up 'H NMR (400 MHz, DMSO-d): 6 8.63 (s, 1H), 8.27 (bs, 1H), 8.20 (d, J= 9 Hz, 1H), 0 (m, 1H), 7.86 (bs, 1H), 7.78 (s, 1H), 7.33-7.25 (m, 2H), 5.81 (bs, 1H), 5.54 (dd, J= 8.8 8.13-8.07 Hz, 1H), 5.28 (dd, J=9.7 Hz, 1H), 4.65 (d, J= 12.4 Hz, 1H), 4.51 ( dd, J= 8.8 Hz, 1H), 4.38 (bs, 1H), 4.03 (m, 1H), 3.97 (s, 3H), 3.92-3.87 (m, 1H), 3.54-3.46 (m, 2H) , 2.68-2.65 (m, 2H), 2.47- (m, 1H), 2.15 (dd, J= 8.6 Hz, 1H), 1.78-1.65 (m, 2H), 1.6-1.12 (m, 17H), 1.25 (t, J= 7.3 Hz, 2.38 3H). 0 (M+Na)", es": 761.2 (M+H). 783.2, MS; YIVY

Aaa; Jha

Compound 877 was prepared using the same procedure described in the example; but with interaction

bromoketone ¢ or with N-iso-propylthiourea is commercially available.

Na | 23 dead A ? = b” m(beobd0 ©O Y 0 oH ¢ Cp 'H NMR (400 MHz, DMSO-d): 8.63 (s, 1H), 8.33-8.23 (bs, 1H), 8.21 (d , J=9.2 Hz, » 1H), 8.04 (d, J= 8.3 Hz, 1H), 7.86 (bs, 1H), 7.77 (s, 1H), 7.35-7.23 (m, 2H), 5.81 (bs , 1H), 5.52 (dd, J= 8.5 Hz, 1H), 5.27 (dd, J=9.2 Hz, 1H), 4.65 (d, J= 11.8 Hz, 1H), 4.51 (dd, J=7.6 Hz , 1H), (bs, 1H), 4.15 (bs, 1H), 4.07-3.98 (m, 2H), 3.97 (s, 3H), 3.88 (d, J=8.9 Hz, 1H), 2.6-2.53 (m, 4.37 2H), 2.47-2.37 (m, 2H), 2.19-2.1 (dd, J= 9.2 Hz, 1H), 1.8-1.64 (m, 2H), 1.63-1.29 (m, 13H), 1.27 and 1.25 (dd, J= 6.5, 6H), 1.23-1.09 (m, 2H).01 (M+H)", es": 772.9 (M-H). 775: i.e. MS; YYVYY

\ TV av 4 Ex. ml 1.17 (Efakm V+) DIPEA mmol) from 11 A) added 5,; ml g VE equivalent) to a stirred solution of methyl chloroformate (mmol) of ©. (6 g) in © ml of 2-amino-thiazolyl (¥£) mmol) of an intermediate of +, TY) at room temperature. The reaction mixture was stirred for six and a half hours before it was concentrated in the desired CHCl 0 as described in the carboxylic acid vacuum. Then the resulting isolated raw material was hydrolyzed to yield

AVA until the compound yield TE in the example is 0 g 5

N

N

9 o 4 puncture 8 0 o © : & OH — 'H NMR (400 MHz, DMSO-dq): 5 8.61 (s, 1H), 8.21-8.07 (m, 2H), 7.61-7.38 (m , 2H), 7.26 (d, J= 6.4 Hz 1H), 7.19-7.10 (m, 1H), 5.6-5.47 (m, 2H), 5.27 (dd, J= 9.2 Hz, 1H), 4.634.533 00 ( m, 1H), 4.47 (d, I= 7.9 Hz, 1H), 4.13-4.04 (m, 1H), 3.93 (s, 3H), 3.92-3.87 (m, 2H), 3.79 (s, 3H), 2.42 -2.3 (m, 2H), 2.17 (dd, J= 9.2 Hz, 1H), 1.81-1.68 (m, 2H), 1.63-1.29 (m, 16H), 1.23-1.10 (m, 2H).

MS; es: 791.1 (M+H), es: 789.1 (M-H).

Vfa

Example 4e by making use of are and following the conditions described above in the corresponding lds example which holds substituents. Then carbamate is an intermediate compound of 3-cisobutyl chloroformate

AVA Crude isolate to desired compound 0 3 ort OY

CHy N, RK. 9 ° : z paracentesis fo © 6 + ¢ OH ss o' H NMR (400 MHz, DMSO-d): 8.62 (s, 1H), 8.47-8.27 (bs, 1H), 8.18 (d, J= 8.6 Hz, 1H), 7.69-7.60 (m, 1H), 7.6-7.51 (m, 1H), 7.28 (d, J= 6.7 Hz, 1H), 7.28-7.19 (m , 1H), 5.7-5.6 (m, 1H), 5.52 (dd, J= 8.3 Hz, 1H), 5.27 (dd, J=9.8 Hz, 1H), 4.63 (d, J= 11.8 Hz, 1H), 4.53 -4.44 (m, 2H), 4.1-3.99 (m, 1H), 4.04 (d, J= 6.7 Hz, 2H),3.95 (s, 3H), 3.94-3.87 (m, 1H), 2.65-2.53 (mm , 1H), 2.46-2.34 (m, 1H), 2.16 (dd, J= 8.1 Hz, 1H), 2.03-1.91 (m, 1H), 1.79-1.09 (m, 20H), 0.95 (d, J= 1 6.7 Hz, 6H).

MS; es”: 833.2 (M+H)”, es: 831.2 (M-H)”.

YYVYY

Example vo Synthesis of compound 008 Prepare the following saturated macrocycle compound 08 (see Table 3) Teas from derivative YY using the same chemical conditions as described in example TE 0

BocHN EN N, — \ chemical conditions as ra \ 0 ivy 5 l love ِ 0 £ oY 3 2 be “mu 2 A 0 6 compound . 'H NMR (400 MHz, DMSO-d): 8.47 (s, 1H), 8.16 (d, J= 10 Hz, 1H), 8.15-8.07 (m, 1H), 7.82-7.63 (m, 2H ), 7.53-743 (m, 2H), 7.33-7.22 (m, 1H), 7.13 (d, J= 7 Hz, 1H), 5.77-5.65 (m, 1H), 4.62-4.52 (m, 2H), 4.50-4.4 (m, 1H), 4.20-4.10 (m, 1H), 3.94 (s, 3H), 3.89-3.83 (m, 1H), 2.59-2.53 (m, 1H), 2.48-2.4 (m, 1H ), 1.79-1.00 (m, 25H).

MS; es’: 735.2 (M+H)”, es’: 733.2 11 .

VE ire Example 064 Synthesis of the compound except using Yo. Compound 9.9 was prepared using the same procedure as described in Example 4. Commercially available (see table Neacetylthiourea

J

mh 13 h 9 Q OX 0 4 8 0 © for l : om '" H NMR (400 MHz, DMSO-d¢): 8 8.53-8.41 (m, 2H), 8.20 (d, J= 9.2 Hz, 1H), 7.68 (bs, 1H), 7.68 (bs, 1H), 7.27 (dd, J= 9.2 Hz, 1H), 7.15 (d, J= 6.4 Hz, 1H), 5.67 (bs, 1H), 4.65-4.5 (m, 3H), 4.44-4.37 (m, 1H), 4.21-4.13 (m, 1H), 3.96 (s, 3H), 3.99-3.86 (m, 1H), 2.62 -2.39 (m, 2H), 2.24 (s, 3H), 1.78-1.67 (m, 3H), 1.67-1.01 (m, 22H).

MS; es’: 798 (M+Na)’, es: 777 (M+H).” 01 th 1

Example ® a Synthesis of compound AY as compound 91 using the same procedure described in Example Yo except for the use of commercially available Neethylthiourea (see Table 4). ~~ 5 CH,0 N | 2 i Q Q or 5 ,% . jd; N 0 v 0 0 om : 'H NMR (400 MHz, DMSO-dy): 5 8.47 (s, 1H), 8.29 (bs, 1H), 8.20 (d, J= 9.2 Hz, 1H ), (bs, 1H), 7.87 (s, 1H), 7.77 (s, 1H), 7.32 (dd, J= 9.2 Hz, 1H), 7.14 (dd, J= 6.7 Hz, 1H), 5.78 8.09 (bs. 1H), 4.58 (dd, J=8.1 Hz, 2H), 4.43 (bs, 1H), 4.18-4.12 (m, 1H), 3.97 (s, 3H), 3.87 (d, J= 8.9 Hz, 1H), 3.55-3.46 (m, 2H), 2.63-2.53 (m, 1H), 2.47-2.41 (m, 1H), 1.78-1 (m, 25H), 1.25 (t, J= Hz, 3H). 1 7.3 MS; es”: 763.1 (M+H)”, es: 761.1 1-11(

VEY

—avre Ex. 911 Synthesis of the compound by exception using Yo. Same procedure as described in Ex. al. Compound 91 was prepared using commercially available 3) N-iso-propylthiourea (see table

J i. wears | 5_2 9

Q

0 % 8 0 © oO - : om o 'H NMR (400 MHz, DMSO-d): 5 8.47 (s, 1H), 8.29-8.19 (m, 1H), 8.19 (d, J= 9.2 Hz, 1H), 8.09-8 (m, 1H), 7.83 (bs, 1H), 7.74 (bs, 1H), 7.31 (d, J= 8 Hz, 1H), 7.14 (d, J= 6.4 Hz, 1H ), 5.76 (bs, 1H), 4.64-4.53 (m, 2H), 4.44 (bs, 1H), 4.22-4.09 (m, 3H), 3.97 (s, 3H), 3.87 (d, 1-6

Hz, 1H), 2.63-2.58 (m, 1H), 2.46-2.41 (m, 1H), 1.79-1.1 (m, 24H), 1.27 and 1.26 (2xd, J= 6.5 Hz, 6H). 1

MS; s: 777 (M+H)", es: 775 (M-H)..

Yvy

VEY

1 Example VAT compound synthesis

A

6 a b = free Q CAN coon

N

BOCNH,, 0 = tes, 0

Pa'" H NMR (400 MHz, DMSO-dq): 6 (ppm): 8.62 (s, 1H), 8.13 (d, J=9.2 Hz, 1H), 7.64-7.54 (m, 2H), 7.47 (d , J= 2.6 Hz,1H), 7.16 (dd, J=9.2 Hz, 2.2 Hz, 1H), 7.03 (d,J]=6.0 Hz,1H), 5.63 (s, 1°H), 5.52 (q, J= 9.9 Hz, 1H), 5.26 (t, J=8.9 Hz, 1H), 4.62 (d, J= 11.45 Hz, 1H), 4.45 (dd, J=9.2, 8.27 Hz, 1H), 4.02 (m, 1H), 3.93 (s, 3H), 3.7 (dd, J= 7.6 Hz, 1H), 2.66 (s, 3H), 2.55-2.65 (m, 1H), 2.35-2.45 (m, 1H), 2.17 (q , J= 8.6 Hz, 1H), 1.65-1.75 (m, 2H), 1.5-1.35 (m, 7H), 1.15 (s, 9H).

MS: 705 (M+1), 703 (M-1). a

Vet 1 Example (VAY) compound synthesis

A

N

N, /

Oo9iCOOH

N

BOCNH,, 0 60 sq' H NMR (400 MHz, DMSO-de): & (ppm): 8.62 (s, 1H), 8.15 (d, at 8.9 Hz, 1H), 7.62 (s, 1H), 7.49 (s, 1H), 7.19 (dd, J= 9.2, 2.2 Hz, 1H), 7.02 (d, 12 5.4 Hz, 1H), 5.64 (5, 1H), 5.52 (q, J= © 9.9 Hz, 1H), 5.26 ) J= 9.2 Hz, 1H), 4.63 (d, J=11.44 Hz, 1H), 4.45 (t, J= 9.2 Hz, 1H), 3.94 (s, 3H), 3.9-3.8 ( m, 1H), 2.7-2.55 (m, 1H), 2.4-2.3 (m, 1H), 2.18 (q, J=8.9 Hz, 1H), 1.75-1.65 (m, 2H), 1.5-1.2 (m, 7H), 1.14 (s, 9H).

MS: 705 (M+1), 703 (M-1). a

Yeo

FA Example (VIA) Compound Synthesis No ~ 9 0 Bah you want Za

Oh

, 0 2: 5 o Free 'H NMR (400 MHz, DMSO-dq): ppm: 9.55 (s, 1H), 8.63 (s, 1H), 8.43 (s, 1H), 8.13 (d, 1- 9.2 Hz, 1H), 7.66 (s, 1H), 7.46 (s, 1H), 7.32 (d, solution to 2.6 Hz, 1H), 7.10-7.07 (m, 2H), -5.64 5.54 » (m, 1H), 5.59-5.48 (m, 1H), 5.33-5.23 (m, 1H), 4.73-4.61 (m, 1H), 4.45 (dd, J=7.5, 9.1 Hz, 1H), 4.09 -4.00 (m, 1H), 3.92 (s, 3H), 3.93-3.83 (m, 1H), 2.67-2.55 (m, 2H), 2.53-2.43 (m, 1H), 2.42- 2.31 (m, 1H) , 2.23-2.12 (m, 1H), 1.81-1.66 (m, 2H), 1.52-1.42 (m, 2H), 1.42-1.25 (m, 6H), 1.21 (s, OH).

MS: 689.3 (M+1), 687.3 (M-1). 6 Al va Example (VYY) compound synthesis

Oo~

Q

0 9 8 and so on of 0 {

H 90 'H NMR (400 MHz, DMSO-dy): 8 (ppm): 9.70 (s, 1H), 8.64 (s, 1H), 8.26 (s, LH), 8.14 (d, 1- 9.2 Hz, 1H), 7.45 (s, 1H), 7.30 (d, J= 2.5 Hz, 1H), 7.14-7.06 (m, 2H), 5.60-5.54 (m, 1H), 5.58- © 5.48 (m, 1H ), 5.31-5.23 (m, 1H), 4.71-4.62 (m, 1H), 4.49-4,40 (m, 1H), 4.08-3.99 (m, 1H), 3.92 (s, 3H), 3.92-3.84 (m, 1H), 2.69-2.54 (m, 2H), 2.53-2.46 (m, 1H), 2.42-2.31 (m, 1H), 2.37 (5, 3H), 2.22-2.13 (m, 1H), 1.81 -1.64 (m, 2H), 1.54-1.42 (m, 2H), 1.42-1.27 (m, 6H), 1.22 (s, 9H).

MS: 703.3 (M+1), 701.3 (M-1). A yey 4 0 Example (vrv) compound synthesis

Oo 0 9 0 0 0 N H

SLY

0 N Sy 0 Free 'H NMR (400 MHz, DMSO-de): ppm: 8.75 (m, 1H), 8.62 (s, 1H), 8.06 (d, J=9.2 Hz, 1H), 7.88-7.87 (m, 1H), 7.48 (s, 1H), 7.28 (d, 1-6 Hz, 1H), 7.05-7.00 (m, 2H), 6.64-6.63 (m, ° 1H), 5.62-5.58 (m, 1H), 5.55-5.49 (m, 1H), 5.28-5.24 (m, 1H), 4.64-4.61 (m, 1H), 4.48-4.44 (m, 1H), 4.07-4.03 (m , 1H), 3.91 (s, 3H), 3.92-3.85 (m, 1H), 2.67-2.54 (m, 2H), 2.53-2.45 (m, 1H), 2.41-2.34 (m, 1H), 2.20 -2.14 (m, 1H), 1.75-1.69 (m, 2H), 1.50-1.43 (m, 2H), 1.41-1.32 (m, 6H), 1.17 (s, 9H).

MS: 689.3 (M+1), 687.2 (M-1). 1

VEA

4 1 Example (Vv 0 9) compound synthesis

J ory

Q

9 L > Lp © > i < OH 'H NMR (400 MHz, DMSO-dq): 8 8.50 (s, 1H), 8.19 (s, 1H), 8.17 (s, 1H), 8.11-8.00 (m, 1H), 7.88-7.77 (m, 1H), 7.73 (s, 1H), 7.25 (d, J= 8.6 Hz, 1H), 6.93 (d, at 6 Hz, 1H), 5.89-5.68 (m, © 1H), 4.62 (d, 1211 Hz, 1H), 4.53 (dd, J= 8.3 Hz, 1H), 4.16-4.07 (m, 1H), 3.96 (s, 3H), 3.88 (bd, J = 9.5 Hz, 1H), 3.53-3.43 (m, 2H), 2.63-2.51 (m, 1H), 2.46-2.36 (m, 1H), 1.81-1.62 (m, 2H), 1.60- 1.01 (m, 15H ), 1.24 (t, I= 7.4 Hz, 3H), 1.17 (s, 9H).

MS; es: 751.1 (M+H)", es: 749.1 (M-H).

YYVY

Ved £Y Example (VF) compound synthesis

Pa N 2 Pa 2 0 Lala GR 0 N 5 0

FH

IH NMR (400 MHz, DMSO-dq): 5 (ppm): 8.62 (s, 1H), 8.54 (s, 1H), 8.04 (d, J=9.2 Hz, 1H), 7.70 (s, 1H), 7.43 (5, 1H), 7.24 (d, J= 2.6 Hz, 1H), 7.05-6.98 (m, 2H), 5.57-5.54 (m, 1H), 5.55-5.48 (m, 1H), 5.28-5.24 (m , 1H), 4.63-4.59 (m, 1H), 4.47-4.43 (m, 1H), 4.13-3.99 (m, 1H), 3.90 (s, 3H), 3.92-3.83 (m, 1H), 2.67-2.55 (m, 2H), 2.53-2.46 (m, 1H), 2.43-2.31 (m, 1H), 2.22- 2.15 (m, 1H), 2.15 (3H), 1.75-1.70 (m, 2H), 1.51-1.42 (m, 2H), 1.41-1.28 (m, 6H), 1.17 (s, 9H).

MS: 703.2 (M+1), 701.3 (M-1).

Yoo

EY Example (Vv rv A) iS yall Synthesis Oo p Q 0 Z 0 0 N i

PY Dd oH 0 : 0

By, 'H NMR (400 MHz, DMSO-d): 6 (ppm) 8.64 (d, J=2.5 Hz, 1H), 8.62 (s, 1H), 8.04 (d, J= 9.2 Hz, 1H), 7.39 (s, 1H), 7.24 (d, J= 2.5 Hz, 1H), 7.04 (d, J= 6.0 Hz, 1H), 6.99 (dd, 2.2, 9.2 Hz, ° 1H), 6.43 (d, J=2.2 Hz, 1H), 5.62-5.57 (m, 1H), 5.56-5.47 (m, 1H), 5.31-5.22 (m, 1H), 4.65-4.56 (m, 1H), 4.45 (dd, J= 7.6, 8.9 Hz, 1H), 4.07-4.00 (m, 1H), 3.90 (s, 3H), 3.88-3.84 (m, 1H), 2.68- 2.56 (m, 2H), 2.54-2.43 (m, 1H), -2.42 2.31 (m, 1H), 2.34 (s, 3H), 2.24-2.14 (m, 1H), 1.80-1.64 (m, 2H), 1.52-1.43 (m, 2H), 1.43-1.27 (m, 6H ), 1.18 (s, 9H).

MS: 703.2 (M+1), 701.2 (M-1). ye

YYVY yo 4 4 Example Synthesis of the compound (5,7).

Az “OO 0 0 0 N: 8

SA + Se

A©.

Va '{f NMR (400 MHz, DMSO-d): 5 (ppm): 8.62 (s, 1H), 8.10 (d, J=9.2 Hz, 1H), 7.57 (5, 1H), 7.49 (s, 1H ), 7.35 (d, 1- 2.2 Hz, 1H), 7.09-7.03 (m, 2H), 5.65-5.61 (m, 1H), 5.55-5.49 (m, 1°H), 5.28-5.24 (m, 1H) , 4.62-4.57 (m, 1H), 4.49-4.45 (m, 1H), 4.08-4.01 (m, 1H), 3.93 , 3H), 3.92-3.86 (m, 1H), 3.2-3.14 (m, 1H) , 2.65-2.56 (s, 1H), 2.53-2.47 (m, 1H), 2.42-2.35 (m, 1H), 2.22-2.15 (m, 1H), 1.79-1.68 (m, 1H), 1.5-1.43 ( m, 2H), 1.41-1.28 (m, 12H), 1.18 (s, 9H).

MS: 748.2 (M+1), 746.2 (M-1).

YY VY

Voy to Example (VY) compound synthesis 0

A N N

\

Q

0 0 0 N8

SLY - 0 i 5 0 'H NMR (400 MHz, 101150-0(: 6 (ppm): 8.64 (s, 1H), 8.10 (d, J=9.5 Hz, 1H), 7.83-7.76 (m, 2H) , 7.60 (s, 1H), 7.44-7.42 (m, 1H), 7.18-7.01 (m, 2H), 5.56-5.49 (m, 2H), 5.29-5.24 (m, ° 1H), 4.66-4.63 (m , 1H), 4.47-4.42 (m, 1H), 4.28 (s, 3H), 4.06-4.02 (m, 2H), 3.94 (s, 3H), 3.93- 3.86 (m, 1H), 2.66-2.55 (m , 2H), 2.42-2.31 (m, 2H), 2.22-2.14 (m, 1H), 1 .79-1.65 (m, 2H), 1.52- 1.27 (m, 7H), 1.22 (s, 9H).

MS: 703.2 (M+1), 701.3 (M-1).

YYVY

Voy 6 6 Example (307) compound synthesis

N

A° N. 8 Free Qo 0 Oily AY : 0 0

Y, 'H NMR (400 MHz, DMSO-d): 5 (ppm): 8.46 (s, 1H), 8.06 (d, J= 9.2 Hz, 1H), 7.57 (s, 1H), 7.49 (s, 1H) ), 7.34 (m, 1H), 7.14-7.05 (m, 2H), 5.63-5.58 (m, 1H), 4.66-4.61 (m, 1H), 4.54- 4.44 ° (m, 2H), 4.23-4.18 ( m, 1H), 3.93 (5, 3H), 3.92-3.88 (m, 1H), 3.21-3.14 (m, 1H), 2.44-2.33 (m, 1H), 1.35 (d, J= 7 Hz, 6H) , 1.73-1.01 (m, 26H)

MS: 762.0 (M+1), 759.9 (M-1). No

Voi 417 Example (9097) Compound Synthesis W.7

Q

0 0 9 N i

JY

IH NMR (400 MHz, DMSO-dq): 5 (ppm): 8.46 (s, 1H), 7.98 (d, J=8.9 Hz, 1H), 7.91-7.89 (m, 2H), 7.23-7.21 (m, 2H), 7.07-7.00 (m, 2H), 6.35-6.32 (m, 2H), 5.64-5.58 (m, 1H), 4.65-4.61° (m, 1H), 4.53-4.47 (m, 2H), 4.24 -4.19 (m, 1H), 3.90 (s, 3H), 3.86-3.84 (m, 1H), 2.40-2.33 (m, 1H), 1.73-1.01 (m, 26H).

MS: 702.0 (M+1), 699.9 (M-1).

VY

Yeo fev The example is (AY0) the compound but in step g use YE using the same procedure described in Mithqal. ( AYo ) Product of the compound N-cyclopropylthiourea m 0 N N 9 2 0 l apm l 2 0 8 0 3 & OH — 1 'H NMR (400 MHz, y-1150): ‎ 6 8.55 (bs, 1H), 8.38 (bs, 1H), 8.02 (d, J= 8.9 Hz, 1H), 7.53 (s, 1H), 7.42 (s, 1H), 7.28 (d, solution 1.6 Hz, 1H), 7.12 (bs, 1H), 7.04 (d, J= 8.3 Hz, 1H), 5.52- 5.37 (m, 2H), 5.34-5.13 (m, 1H), 4.75-4.64 (m, 1H) , 4.41 (d, all 15.9, 8.9 Hz, 1H), 4.33-4.18 (m, 1H), 4.09-3.98 (m, 1H), 3.98-3.83 (m, 1H), 3.91 (5, 3H), 2.57-2.43 (m, 1H), 2.06 (s, 3H), 1.78- 1.18 (m, 19H), 1.16-1.12 (m, 1H), 0.78-0.72 (m, 2H), 0.61-0.54 (m, 2H 01

MS; yen 773.4 (M+H)", es: 771.5 (M-H)".

YYVY you [ to CQ _

KSCN

HN

0 5 above H H is the intermediate compound

JA

H.N.N

0.5 ml overnight under a vacuum before use. Then KSCN Yd was added dropwise to a solution of 0,000; g benzoylchloride (7.07; mmol) of and stir the solution 0. At GE acetone mL of YO in KSCN and mmol of TY) 8 mL (£750 mmol) of 1" milk in an ice bath for an hour and a half; then added 0 to a light yellow opaque mixture.The reaction mixture was stirred for (dats 1 drop of cyclopropylamine of (Use mM VST) 0 μl further 0 ½ hr at 0°C; then 1 min was added. At that time the reaction mixture was stirred at room temperature for 71 mL of Tov cyclopropylamine and the reaction mixture was stirred into a mixture of Guay (HPLC) It was determined that the reaction was complete with the light yellow solid filtered out and washed 0 It was stirred for five minutes (H.0/ 2a ye gm of the intermediate compound. 1 several times B 11:0 and dried in vacuo to produce a standard and heated 1, its concentration is NaOH ml of a solution of 501 gm From intermediate in AY B: Commented to completion of transformation of intermediate into product HPLC min. 11 device indicated with rewind for 0 times FtOAc annealed and extracted in NaCl — and saturate cad yall and the solution was cooled to a temperature twice and with brine water once; it was dried over HO after mixing it with EtOAC and washing the yo extracts

YIVY

Yov was filtered and evaporated to produce the crude product as a white-yellowish solid. N-cyclopropylthiourea (10+2steps<'H NMR (400 MHz, DMSO-de): 7.92 (bs, 1H), 7.61 (bs, 1H), 7.13 (bs, 1H), 2.39 (bs, 0 1H), 0.67-0.63 (m, 2H), 0.51-0.44 (m, 2H),

MS; es”: 116.9 (M+H)”, es: 114.8 (M-H)”.

WEY Example (AYV) compound synthesis. for Ly 0 SL

OY

LO o vn and Wp ay 0 until the end of step h; But in TE using the same procedure described in the example

AAYY) to produce the compound N-cyclopentylthiourea step g used

Yvy

11 d love and

HN

use

LA Tomy to a solution of 0.0 ml cyclopentylamine ml (14 mmol) of 17 aul a: and stir DIEA — follow CH,CL, 001 ml of 9.51 t-butyl isothiocyanate ? mmol) and EtOAC—reaction mixture was washed at room temperature for 2 hours. The mixture was diluted twice. and B 11.0 twice with brine water, NaHCO; — twice” and saturated 701 O1N1H with an anhydrous 0 acid concentration; MgSO was filtered and evaporated to dryness, one time. The organic layer was dried over in the form of a white solid (t-butyl-cyclopentylthiourea rate of 7.71 g of production.)/47 production-concentrate. HCL was heated in 71 lbs mL of t-butyl-cyclopentylthiourea 00 g of B:dissolve the dark yellow solution with moderate reflux. after £0 minutes; Allow the reaction mixture to cool. the volume is concentrated to about half under reduced pressure; It was cooled in ice and made a solid and saturated base. The product was extracted in NaHCO, until the pH reached 9.5 using after mixing it == 11.0 twice and with once brine water EtOAc three times; The EtOAc extracts were washed, filtered, and evaporated to dryness (SLO 10850, one). The organic layer was dried over in the form of a beige solid. N-cyclopentylthiourea 17 g of crude was obtained in a ratio of 0/40. After flaking: 7,748 g hexane/EtOAC by grinding the raw material in a mixture of (74 = z) in the form of a white solid (modified N-cyclopentylthiourea from No

'H NMR (400 MHz, 1180-4): 7.58 (bs, 1H), 7.19 (bs, 1H), 6.76 (bs, 1H), 4.34 (bs, 1H), 1.92-1.79 (m, 2H), -1.66 1.55 (m, 2H) 1.55-1.30 (m, 4H).

MS; es”: 144.9 (M+H)”, es”: 142.8 (M-H)”. (Na salt) “H NMR (400 MHz, DMSO-d): 8 8.02 (d, J= 9.2 Hz, 1H), 7.09 (d, all 6.4 Hz, 1H), 7.76 (s, 1H), 7.44 (bs, 2H), 7.27 (d, at 1.9 Hz, 1H), 7.11 (d, I= 7.0 Hz, 1H), 7.03 (d, ]=92©

Hz, 1H), 5.48 (dd, J= 18.4, 9.9 Hz, 1H), 5.43 (bs, 1H), 5.15 (dt, J= 17.8, 7.63 Hz, 1H), 4.7 (bs, 1H), 4.49-4.34 (m, 2H), 4.34-4.25 (m, 1H), 4134.03 (m, 1H), 3.99-3.86 (m, 1H), 3.90 (s, 3H), 2.58-2.44 (m, 1H), 2.42-2.32 (m, 1H), 2.15-1.93 (m, 4H), 1.83-1. 14 (m, 24H), 1.14-1.12 (m, 1H).

MS: es” 801.4 (M+H)”, es”: 799.3 (M-H). —atV Example Ve (ATH) \“or I~ 0 OY) Y

Ro tb 'H NMR (400 MHz, 01450-4): 5 8.22 (d, J= 6.0 Hz, 1H), 8.02 (d, J= 9.2 Hz, 1H), 7.75 (s, 1H), 7.46 (s, 1H), 7.44 (s, 1H), 7.27 (d, J= 1.9 Hz, 1H), 7.11 (d, J= 6.7 Hz, 1H), 7.02 (dd, J= 9.5, 1.9 Hz, 1H) , 5.54-5.41 (m, 1H), 5.44 (s, 1H), 5.14 (dd, J= 15.9, 9.9 Hz, 1H), 4.75-4.66 (m, Veo A

h 1H), 4.48-4.34 (m, 2H), 4.34-4.26 (m, 1H), 4.12-4.02 (m, 2H), 3.90 (s, 3H), 2.57-2.46 (m, 1H), (m, 3H), 2.12-1.95 (m, 4H), 1.82-1.20 (m, 20H), 1.13-1.02 (m, 1H). 2.42-2.31 MS: es”: 787.4 (M+H)' , es: 785.4 (M-H).. Example a Y 0 compound (AYA) l] . goes 7 mm 0 AN YT !x0 = 0 and ak OH 5 —_— (d, I= 9.2 Hz, 1H), 7.86 (bs, 1H), 7.78 (d, 1-3 8.02 AH’” H NMR (400 MHz, DMSO-de):

Hz, 1H), 7.43 (s, 2H), 7.27 (d, J= 2.2 Hz, 1H), 7.12 (d, J= 6.9 Hz, 1H), 7.03 (dd, - 9.2, 1.9 Hz, 1H), 5.57 -5.40 (m, 1H), 5.40 (s, 1H), 5.26-5.17 (m, 1H), 4.70 (bs, 1H), 4.52-4.35 (m, 2H), 4.29- 4.23 (m, 1H), 4.18 -4.00 (m, 1H), 3.90 (s, 3H), 3.87-3.65 (m, 1H),2.42-2.32 (m, 1H), 2.19-2.10 (m, 101H), 2.07-1.96 (m, 3H ), 1.82-0.95 (m, 28H).

MS; es”: 815.4 (M+H)”, es: 813.4 (M-H).

VIA

478 Example Full-length NS3-NS4A heterodimer fluorogenic assay Full-length NS3-NS4A Heterodimer Protein at the © end of the enzymatic polymerization reaction NS2-NSSB Non-coding region is cleaved In reverse transcriptase-polymerase chain reaction (RT-PCR) vector 5

RNA using (Invitrogen by Jaan Invitrogen) 3m (Registered Trademark) (provided by 1b HOV genotype extracted from the sera of a person infected with the City of Montreal “Hopital St-Luc Hospital St. Louis Dr. Bernard Willems Bac (HTa particle tag B208 in a baculovirus expression vector 0 containing vector context - Gibeo/BRL (obtain from Gibco/BRL pFast-Bac™) contains An N-terminal cleavage at the nitrogenous terminus YA was labeled on a region with a context code for it from 0106 Nakanta”©. A bacillus hesylvirus expression system of the Bac-to-Bac type was used to produce Gibco BRL (obtained It was obtained from Gibco/BRL Bac-to-Bac™ (trademark) from 01 cells by exposing His NS3-NS4AFL recombinant bacillus virus. Protein 00 was expressed at 7 lM. 17-01 for infection with a recombinant bacillus virus at a multiplicity of infection reaching 1 during expression to produce an authentic autoproteolysis protein complex and true autohydrolysis of the FL protein (termed full length NS3-NS4A from non covalent) fixed and non-covalent 1 h by centrifugation at 4 C. TE EA and harvesting of post-infected cell culture after mM; pH 000 concentration NaPO, homogenized cell pellets in -1 mMs pB-mercaptoethanol concentration 756 (w/v); glycerol 3 6 i Dla from His-NS3 -NS4AFL then extracted the protease in the presence of a mixture of protease inhibitors 1410 (Triton X-100 V + += aS) with a concentration of 71.9 and Triton NP-40 chelate using

YIVYY

1" and after separation by ultracentrifugation, molecular DNase and 1.5 concentration of NaCl were treated

Four-fold soluble Ni was fed to a chelation column using Adal diluted Pharmacia obtained from Pharmacia. Hi-Trap Ni-chelating column (SDS-PAGE) determined by LS analysis ) 78+ in a pure solution with a ratio of more than His-NS3-NS4AFL, at His-NS3-NS4AFL reaching 5660 mmol. and store ©0 with a graduated concentration of imidazole M with glycerol ¢V,0 mM; Its pH is © 0 at a concentration of sodium phosphate m in “=

NP-40 5 ¢ — ya 1,709 imidazole 5 concentration 0,* mol NaCl (w/v) 0 dissolved on ice and diluted immediately before use./0.1 mM 3S 0 in 11151101 concentrated His-NS3-NS4AFL and the protease activity was titrated to n-dodecyl-B-D-maltoside at a concentration of 7*9 molecules; pH ye mmolecular. Substrate was incubated internally hydrated 1 with TCEP concentration of 70.01 (w/v); (context anthranilyl-DDIVPAbu [C(0)-0]-AMY (3-NO,) TW-OH Microparticle concentrations of His-NS3-NS4AFL in the presence of different concentrations of the inhibitor with 1) Discrimination No.: final of 78.75 and nanoparticle DMSO anhydration for £0 min at 77 °C. The concentration of 5 molecules did not increase; And its pH is 8,8. The fluorescence of the 1 MES reaction product was monitored by the addition of Vo

LS-50B type L = +©B nitrogen tip fluorometer on a fluorometer equipped with a Perkin-Elmer plate reader of 937 eyes obtained from the Perkin-Elmer Company. nanometers) EVY nanometers; Emission wavelength: ¥YO excitation wavelength: 96-wellplate reader (Ish) The inhibition percentage was calculated using the following equation: [001 X (go gp fluorescence ge BI) (i ge ser-alkylation Rolftla) -001 for Hill model data compatible with a non-linear curve and plotting a non-linear curve using (ICs0) inhibition versus concentration; effective concentration for 7596 subjects treated Statistical Software System Inc. Statistical Software System SAS Software L1

Vir North Carolina Cary SAS Institute, Inc. North Carolina (N.C.) £4 Example HCV recombinant in NS3 protease NS3 protease radiometric radiometric titer HCV in NS3 The substrate used for radiosampling of the protease cleaved serine and systeine between oY cysteine moieties (recognition context no.: DDIVPC-SMSYTW) natural cleavage site Y (recognition context No.: DDIVPC-SMSYTW enzyme pathway. The context corresponds to .proline — P2 was incubated in systeine where the cysteine 3 moiety was replaced (distinguishing context 6m: ?) and the substrate of DDIVPC-SMSYTW from Peptide (context tag no.: “) with biotin-DDIVPC-SMS ['*I-Y] TW from tracer tracer 01 recombinant protease 1153 in the absence or presence of inhibitors. Separation of the substrate from the products and a mixture was performed. Titration and then by filtration. avidin — coated with agarose beads by adding agarose beads (context highlighting number: ;) contained in the filtrate (with inhibitor 5 [BLY] TW) and use the amount of product for substrate shift and the percentage of inhibition. A gia) to calculate the ratio A:ve reactants from Life Technologies UltraPure (Tris-HCI and Tris obtained from BSA and 5 sigma) “UltrapPure (glycerol) obtained from Life Technologies (CH), SO “Pierce from TCEP and obtained from Pierce. Sigma" (Registered Trademark) Trademark Ale) of Anaemia NaOH Corporation and Aldrich® Aldrich® (Registered Trademark Anachemia® Registered) Y. ¢V,0 with concentration © Molecular pH Tris-HCI Calibrator: mg/mL 1 BSA 77 (w/v); CHAPS (w/v); 7700 glycerol Molecular 1 before use directly from the mother solution, its concentration is TCEP Molecular (1 was added at a concentration of 00 pf yg in water). Micromolecular (from YO (context highlighting number: ?) at final concentration DDIVPC-SMSYTW delivered to the substrate: stored at = 07°C to avoid oxidation). [117-]000-001700-5345]'7:i (discrimination context no: 7)) (final concentration is about © nanoparticle). 1 nanoparticle (from a 15M YO solution; final concentration of HCV Protease 1883 L at glycerol/,# mM; pH 00 at sodium phosphate standard at 10.001 mM ©.; 170-40 at mM DTT; 01 NaCl concentration 0b: Method 00 Each well contained: Lue 176 in polypropylene Calibration performed in a plate of 0 μl of substrate/tracer in titrant buffer; 00 μl + Inhibitor in 770 011:(:50)/Titrant Buffer; 0

Ab μl of protease 1153 type 0. A placebo (without inhibitor and without enzyme) and a control solution (without inhibitor) were also prepared on the same calibration plate. ve at 610 min. The enzymatic reaction began with the addition of the enzyme solution and the incubation of the calibration mixture. For a standard time to quench the reaction, 1070©, its concentration is NaOH 1 μl of 010 M, with gentle stirring. A microliter of agarose beads coated with B:Eh (obtained from Company 01) was added enzymatically and Y. of MADP BN 11180100 plate was added in a Pierce® filter plate. registered trademark) and transferring the “Millipore” mixture under the trade name Millipore (registered trademark) MADP NGS 10 minutes at 77°C with moderate stirring. 610 calibration dripped onto a filter plate and incubated for

YYVYY

Vie plates were filtered using a Millipore filter (registered trademark) and transferred + μl of Millipore’s MultiScreen Vacuum Manifold and 01 μl of fluorescence fluid containing lie AT filtered into a non-slip plate. transparent in it for each eye. scintillation fluid Packard® Top and the filtrate was counted on a Packard apparatus (registered trademark) for one minute. 271 liquid protocol ZT using the Top Count liquid method The percentage of inhibition was calculated using the following equation: [001 X (in for the solution Aan Edmir, the census around SSCH) / (the census for the Daatla solution) [-0 is non-linear, consistent with the Hill model for inhibition data against concentration and only Fade and drawing (SAS Statistical Software System) using software (ICs) 70+ Effective Concentration of 0 Curry City “SAS Institute, Inc. 0 Statistical Software System Inc. (N.C. North Carolina State “Cary ov Example) Quality titrations determined the quality of the compounds on a different set of enzymes: serine protease, elastase from chuman leukocyte elastase, elastase from human leukocytes, iserine proteases, and alpha-chymotrypsin. Bovine porcine pancreatic elastase One pig pancreas: cathepsin 8 from liver cysteine protease bovine pancreatic a-chymotrypsin

Loe 973 In all cases, a plate-based method of human liver cathepsine B, specific for each enzyme, was used. Each titration involved incubating the chromogenic using a Ye primer-stained substrate with an enzyme inhibitor for 1 hour at room temperature, then substrate was added and performed as measured on a hydrolyzed Ty jE 7700 microplate reader to obtain a conversion of Thermomax (Registered Trademark) UV microplate reader

1107 or a fluorescence plate reader of the type LC © P8 with the trade name Perkin-Elmer (registered trademark) and the substrate concentrations were kept as low as possible compared to Ky to reduce substrate competition. The concentrations of the compounds ranged from (Te) to 1.05 micromoles (Tlie) according to their effectiveness. The final conditions for each calibration were as follows: Tris-HCI at a concentration of mM © and a pH of 118.50 0A at a concentration of 0.0 Cea NaCl at a concentration of 00 mM EDTA “dja” at a concentration of 11 Milli ein 01:(,50) 7 Tween-20 750.0 with; Succ-AAPF-pNA] at 001 µM (marking context #0) and a-chymotrypsin Ve at picomolecular Yor]; Suce-AAA-pNA] at 177 µM and esterase Porcine elastase at 1 mM molecular A [17 5S Succ-AAV-pNA] micromolecular and leukocyte elastase at nanomolecular A concentration]; or NaHPO] at 100 mM concentration and pH 1” 2078 1mM 01:(:50) 77 TCEP 1mM Tween-20 76.01 (7-amino-4-methylcormarine) Z-FR- AMC Vo at a micromolecular concentration and cathepsin 8 8 at a concentration of 0 + nanoparticles (the parent enzyme was active in a buffer solution containing TCEP at a concentration of mM Yee before use). to a flat-bottom plate with an AT eye of polystyrene (Y. Cellwells Corning) using a Biomek liquid handler (obtained from Beckmann). (Beckman £0 μl Tris-HCI buffer) 51 nM, pH A, 118:50, “guia 1 11801 090 mM”; EDTA at a concentration of 1 no

Vy is millimolecular); 1

A at 00 mM concentration; pH Tris-HCI) μl enzyme solution Yo 0.7 mM Tween-20; 1.1 mM EDTA; © 0 mM NaCl from pig pancreas at a £0 nanoparticle concentration); and elastase Ju)

A at 00 mM concentration; Tris-HCI pH) 0 μl of inhibitor solution 0 ° 7N Tween-20 mM;11 000 mM EDTA NaCl v/v). 110 An inhibitor with a concentration of 1.0 mM -? micromolecular 011, (0,50) μL of Yo 01 min from the pre-incubation at room temperature was added and after NayS0, A concentration of #50 mM was added; pH Tris-HCI) substrate solution

Succ-AAA-PNA 1.1 mM EDTA Concentration; Or Molecular; 11001 at a concentration of 8-00 at a concentration of 166 microparticles) to each well and the reaction mixture was further incubated at a precise temperature. After this period of time, the absorbance was recorded on a room 01 UV plate reader of the Thermomax type (marker Registered commercial) < 02 m 1060. Rows of eyes were designated as controls (without inhibitor) and as placebo solutions (without inhibitor and without enzyme).

A pH of 00 mM Tris-HCI by liquid handling agent using 70.07 mM Tween-20; 1.1 mM EDTA; © 0 NaCl and .61 (.50) at a concentration of 715 v/v. All other titrations were performed to determine the quality by the same method. The percentage of inhibition was calculated using the following formula: YX. 001 x effective length))] mi ted -/ longest FTV (/)/ length 17 (((71) and drawing a non-linear curve consistent with Hill’s model for inhibition data versus concentration only (Statistical Software System SAS Using software (ICs) 90 effective focus for

VTA

Cary (SAS Institute, Inc. - Statistical Software System (N.C.) North Carolina 1H Jud

NS3 Protease Cell-based Assay NS3 assay human cell line protease human cell line Huh-7 This assay was performed using 0

IDNA and simultaneous transfection with two hepatoma units derived from a liver tumor

HCV — non-structural polyprotein One of which is a fragment of a non-structural polyprotein

Gp. Concatenated NSSA-NSSB through a cleavage site (TA) fused to the Jay acid protein (NSSB) where ¢(NS3) (termed NS3-NS4A-NS4B-NS5A-(NS5B)-TA. 1155 for the first of 1 of the six amino acids amino 01 iE and the unit (CMV) expresses this polyprotein under the control of the Pru promoter secreted alkaline phosphatase 5 le which is a reporter protein for the scheduled protein tTA-responsive promoter tTA in the presence of a secreted alkaline phosphatase (SEAP) response and the first residue results in the expression of a polyprotein from which mature proteins are secreted

LS) fia Mature viral proteins are believed to form NS3 differentiated by section protease-1 fusion protein tTA and represent the endoplasmic reticulum at the membrane of the endoplasmic reticulum (in Bujard Journal and Bujard Gossen described by Goosen fusion protein p: 581-2908497 997 1 pm); It contains an AY field. Natl. Acad. Sci. USA vol. 11/8 protein sorting requires a transcriptional activator section and a DNA transcriptional activator to bind with and itself. This last section leads to NSSA. Depends on 1153 at the site of section 115588 between -0 will lead to a decrease in the activity of the protein state (lly. SEAP) to the nucleus and activation of the TA gene translocation and the accompanying cytoplasm from the cytoplasm tTA to Isolate NS3 proteolytic activity 3. SEAP decrease in activity

YYVY

To set cellular events AY! cellular activities other than inhibition of the compound's protease 1153; Similar infections were carried out using a residue expressing TA alone and the same residue as determined so that SEAP activity did not depend on the 2193 protease. Calibration method: Growing Huh-7 cells were subjected to 0110-5711 (obtained from ° Life Technologies FCS (fetal calfr serum) with a concentration of 0 for a transforming infection synchronized with two DNA units in the following amounts: V NS3 micrograms + Ov ng SEAP + 800 µL FUGENE (Boehringer Mannheim) per 1 x 01 Hub cells. Five hours later, cells were washed, trypsin-treated, and plated on 96 Lie plates (at a rate of 80,600 cells/eye) 01 containing different concentrations of compounds to be tested, and after incubation for 1 hour, SEAP activity was measured in the medium using a Phospha-Light kit (obtained from Tropics). - (Tropix) And the analysis of the percentage of inhibition of the effectiveness of SEAP in relation to the concentration of the compound was carried out using SAS software to obtain: 50 (effective concentration of (or ve) Tables of compounds (puts) The following tables are examples of compounds according to this invention . Where it was found that all the compounds presented in Tables 1 to 4 are effective in the enzymatic titration described in example EA, and the number accompanied by an asterisk (*) represents the enzymatic activity resulting from the radiometric titration described in example £4 so that the ICs values were less than 0 © Micromolecular. In these enzymatic assays; Use the third classification: A < micromolecular; 1 micromole > 5 > 151 micromole; and © < 010 micromolecular. Several compounds were tested in the specificity assays described in Example 09. They were found to be specific for Protease 1053 and in general; The results of the different quality calibrations were as follows: YYVY

l 01 < HLE microjesine; 058 > 201 µmol; alpha-chymotrycin atreta-"H > 001 micromolecular; cathepsin 8 8 Yoo > micromolecular cathepsin; This indicates that these compounds are Alle specific to the NS3 protease of HOV and are not believed to exhibit serious side effects. In addition; Some of these compounds were tested in the cytometric assay described in 0 Example (©) and found to have activity with a value of 0.36 under 01 µM; Which clearly indicates that these compounds can cross the cell membrane. On the canal face) the Gy compounds of Tables 5A HY were estimated in a cytometric titration and the results are shown in the last column. And in this cellular calibration; The following conventions are used: 1 SA micromolecular; 8 < 1 micromolecular. The following abbreviations were used in the following tables: 0 2045 SPV data; k/u THM = k/u THM unless otherwise indicated dad asterisk * ¢ ¢phenyl :Ph Stert-butyloxycarbonyl :Boc ¢benzyl :Bn 'acetyl :Ac .propyl :Pr

1no table 1 ee N R® 0 x 9 N R 1 COOH 0 275 6 0 lg 0 9 7 l 11 AANA 3 8" 12 10 8 where there is one space group at R' Sal number the double bond between the stereochemistry of the MS 2 bond R' the activity two carbon atoms at the position of the enzyme the two positions "or 01" a - 0 011 01- vs 1 position 0 adjacent to *A TAS A phenyl amide 1 not present “1R position VY adjacent to acid 1 phenyl rag 0 1 non ¢” 1R 3d 9 3a position VY Adjacent to *A TAV,Y phenylamide no

schedule?

RA N RZ

Oy x

Q

0 N 8 1 COOH x 2 |x 7 0

R? A 9 13 Pr 4 A ©2100 Th

R' where there is one stereoisomer at activity MS R% rR” double bond stereochemistry rR? The compound number of the enzymatic L bond

VE position 8 847 H H or 15 position 18 of Bac-11 0 H NH-Boc 707 adjacent to acid 04

A ovo, H H or 15 position IR of Baqam-17 1) H NH-acetyl 9. adjacent to acid 4

B YY, H H Position as N/A 18 or 11-4 NH-Boc Y.o acid adjacent 4 12-OH adjacent 0 4 / H H 14 Position IR Rgm-1 4 OY H NH -Boc Yon adjacent to acid A

A continued - No. R4 R3 double bond Stereochemistry MS R22 R21 Compound activity of the RI bond at the enzyme Position VE OY H NH-Boc 1 0-adjacent 18 Position H OMe VE A 0 (IR alae VE OY H NH-Boc 018 position 144,A phenyl OMe VE 0 adjacent to (IR ssa) E 0 H NH-C(0)-NH-tBu 9 position phenyl OMe VE 0 contiguous to acid OY H NH-Boc YY. phenyl OMe VE 0 adjacent to acid AY H OH 711 4-adjacent 11 position B ove H OMe VE (isomer) adjacent to acid (IR Rgm-1 4 0 10-o0x0 NH-Boc Yi position YiY A phenyl OMe VE 0 adjacent to H NH-Boc Yye not present (IR) position VeV, A phenyl OMe Vt 0 adjacent to Rala NH-Boc 10-3 (combination of AY 41 118-adjacent to the phenyl OMe VE position (0 of the two stereoisomers) adjacent to the acid OY 10-0x0 NH-Boc 1-adjacent 118 to the phenyl OMe VE position 0 to the 0 adjacent to the amide H NH -Ac ARR does not exist (IR) position phenyl OMe VE T “C adjacent to acid (IR Rgam-14 AY H NH-Boc YY). Position OMe VE L Brkhla C adjacent For A amide 1 no

l table? 2c L 9 N 8 1 COOH 0 275 6 npm Mo 7 3 R D where pe) has one interstitial at C number 3 -D- Stereochemistry of the 0-8 MS ! bond R% rR “Enzymatic Activity of Compound BY (NH-Boc ~~ ¥+) for IR or 15 D adjacent to acid (TY), H H 8 16 IA 9 15 14 ,” / : 13 12 10 NH-Boc 07 mm B “IR © adjacent to A 71 Ph OMe amide / * 8 11 10 NH-Boc YY 12 10 3 118 © adjacent to core Ph OMe amide on 8 So NNN 13 9 11 p NH-Boc d ¢IR © adjacent to acid Ph OMe 1NO C 15 13 11 M po Na 14 12 10 9 D 118 0 9 11 HO Yo adjacent acid Ph OMe arti 8 J monk 8 v ZF 14 13 12 D 11 7, 0 NH-Boc 1 adjacent to core Ph OMe amide Tai*A 8 10 la

Z NH-Boc -- 0 b DIR adjacent to acid OMe 0 H 0 2, 11 15 N.

N °, 13 / YY Ty 14 1 © A yyy NH-Ac ~~ YA 9 1 18 © adjacent to acid OEt OMe Tbk 0 11 2 14 p 6 3 12 10 Aam Am A M Ht Tb mm mm mm mm mm mm mm YYVYY

Table 7 © - x

Q

H

N N COOH

06pw

SLL EN ° 1 X, uu 2 === 14 8 4 10

R 13 where the C 4 1 bond is adjacent to the acid ey the activity M binary B ada . 1 < x 5 between A J 4 in position * R * enzymatic compound number 17 and 11 in positions 0 (0 opposite tread CH, H £4)

B adjacent to K CH, H ¢ 0 Y 0 Very for 0 H 7 8 for 7 except 0 § £8 md 0 m / 0 for only

Me

C RY N/A 0 H F 8 YiV,Y Lav 2 0 N/A T V4 £2 A 0 N/A Ed 0

Me

B except © , Y adjacent 0 / £44

Me

C for £5 / £4

Me

A VEY, adjacent Y 3 / £1)

Me

A 77 adjacent 9-5 8-(Me), Zul A

Table E, MBBSBSPSRSAL: LNB ADTCKHKH #_ SASAS - N. Term Q Ne 8 COOH 0 7 . 0 8 L Z 6 Ws, 10 A A 0 xX 14 x12 7 YL 9 Cua The 14-8 bond is adjacent to the acid of Sod) * at position AX member (position GX) MS Enzymatic Activity VY 11 Ya. A L L (CH, 0 CH, ou) Rala 0 Va CH, CH, CH, o.Y 0 NH CH, CH, oY p7 A N(Me) CH, CH, eo. NH a.l 08 CH, CH, N(CO)Me oA 7 0 ee a

\YA + table ee R2 N RE © >

Q o 0 N: i COOH > 6 2 | R! 0 “3 | 0 0

N 7 12 / 14 where the bond 1428 is adjacent to the acid Enzymatic activity MS RY rR Compound number — 6 lm”

C for 0 N(Me), 1) mm" m 5 c *1vo,Y (CFs) OH 1

C 14.) a OMe i.e. “for +-”.

17 Table 08.) RB 9 2 8 COOH 0 % 3 | al

PY 7 0 & 7 8 0 7 b” X 12 0 y X xX & x , 11 3 adjacent to acid 14-R! Where the bond is 2 Potency MS Double bond 22g Xu Temporary Xo R* Number of compound between the two cellular sites

VE and 1 ADB MAA AAA

A Arla Phenyl contiguous CH= Xo H vay

CH= Mor 0=Xn B var, (for all) contiguous sap CH, H Vay —!

B 79 (for all) not found Hb CH, H Vey 4 5 8 (for all) adjacent 0 Rdla CH, H vet

Z

N

B vey 0 (for all) adjacent CH, H Vio

0 - Activity to be continued MS Rr? The double bond, Xie Xo Xo, is the compound between the two cellular sites

VE and 1” 8 (for all) adjacent to CH, H for no H YA “+- B 77 Na N (for all) adjacent to CH, H YA —

B vie,¥ N/A Rap (eeal) CH, H Va \ -S

B vo.) (for all) Not found CH, H VY.

Nee for +- 8 AR | aS N/A (eal) CH, H 7 B N B YY -OFEt (for all) adjacent CH, H VAY

B YA, 7 n/a (seal) CH, H L 5 B 14,7 -OFEt present st (all) CH, H vy ¢

B TAAY 1 J] (for all) adjacent CH, H vie /

A Yeo ¥ ~~ (for all) adjacent to CH; H 71 for 8 (for all) adjacent to 7 for said CH, H VAY oN

Aha follows _ No. Xs Xa 4X0 R* Double bond MS RZ Activity compound between cellular positions "1" and YE except CH, H (for all) adjacent to B TARY a / CH, H via (for all) adjacent to Rala B N rd (geal) CH, H VY. N/A Haem Rala B !1 cH, H AR (for all) N/A B yay 1] Free CH, H 777 (all) adjacent to L3 OD AB and 2 (ed) CH, H 7 N/A 7 ¢ A 8 N / (meal) oH, OH VY € Existing ~ B VAY, 1< CH, H vYo (all) adjacent to EN kick B — CH, H “771 (all) adjacent to B Yor, Y iD) A \

VAY

- Follows the activity MS R * binary aay and R Xo Xo number compound between the two cellular sites

VE GY ee ——

A L,Y -CH,-OMe (for all) adjacent to CH, H the L A 27/7 Me (for all) adjacent to CH, H Vala 8 the R (for all) adjacent cH, H 74 bd > _S

B YAY (for all) does not exist Sub CH, H 77 N 8 (for all) adjacent to 1 l 03. 0 8 ve — 5

A A L Je (to all) adjacent to cH, H 77 0 8 TAY 2 (to all ( to adjacent to) CH, H 77

J

B 71 (for all) adjacent to CH, H “7 7 )

J

B vias (for all) adjacent to ab cH, 0 vre -+ 5

H

B (for all) is adjacent to one within cH, H vr a 0

B no LY oy (for all) adjacent to CH, H balala free N A

VAT

—_— Continued Activity MS R* Double bond Xing of Xo rR? The compound number between the two cellular sites

VE and 1.”

B VARA cr (for all) adjacent to CH, H VYA

J

B No Ph (for all) N/A CH, 10-(R)Me 4 8 A Ph (for all) N/A CH, 10($)Me VE:

B vir) (for all) adjacent to dah CH, H except for +- AA

YAS

A table - mmm mmmm wow aaa 0 N 2 > 9 8 0 N COOH 6 z 8 1 0 0 2 the still 8 10 7 12 14 1732 DZD / mg MN 13 er Asay O, where the 14-8 bond is adjacent to the acid; And the double bond between the two positions A and 3" is in an adjacent position - SS SM TSO SS SSSSSSMM 01 TSMM to the mechanism MS 122 1 132 Cellular compound number a what mm mm mm p hm hm hm hm with mm mm mm mm 8" riel Ne mil H A) + 5 y A ry, vy OEt H n-Pr AY

LY H At

B Va, map > —U!

B Yi © H Avo nt Co

A 1177 08 H "4 A

B 4, OF HCl. ALY p A done 08 H PY Av A oo”

YYVYY

Ma 9 l’ 8 b d B ve. r !-- H Sy Me B ~ rel B !— NH H CI.

AN for B VEV,Y ~ y ~ \_-s N NH H CI.

AY lee B xxx ~ + - H x AY 08 4 8 YIVY

M No. MS R* 8 R* Efficiency H AYE [( Date 8 0 5 OL H Ave R L 8 cm St N 0 A 8 8 to tampon B 'Ne' NS —T > a.

H AVY CL : A B H AYA 5 CL M 1 + P 0 ! — AYA H "RAM B A SP 0 — AY. None B VEV,Y ~~ H 7 5 — AY y J ALT a y yy ! +- a

YAY

Event MS R” R* R* No. L L A L L 8 7 AH except cr H AT / L 8 qr, 08 10-(R) Me CL AYE SS 0 L'come +! p. 0 “his ‘talm <I p

Fo QL” ff 8 4m rt H CL AYA + 0 2 a no

YAA

1 Table es 0 N RZ 9 Pal Tat 6 1 A 8

A 8 None 11 10 12 14 9 gM 13 Where the bond 14-8 is adjacent to the acid. Cellular activity MS Compound number 2 p a p a a ence ya sassa sassa ssssa sassa sassa ssssssss ssssss sssss sssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssssss A 8 8 8 08 H CL a) oo”

B 4 0 H CLL iy p 8 74 5. H no 7 y sa a 0

B TAQ, and H 9.0

NN yd 4 0 th 1

4m to be continued - compound number MS R “R* Rr” cellular activity H CI. 3.1 for vay, 8 y — CJ H CL q.v for 8 y/m H CL wt this 8

pp we A 4.9 to create a ed 0 ay. for ( VY <I H 8 5 y H CL 41) we A 8 1 a and oe oo” B 77 8 H 9 N;—T Y Ng?” H CI. August 16 B Via, >< - <= 0 5 B yoy) 8 H 91 D O) | ]+- 140,Y 08 10-(R) Me CL 411 8 er YAVYY

Va. Context table Highlight context number: >< 1 Highlight context number: <710> 11 Length: <711>

PRT Type: <A11< Member: Industrial Context <71X3< Features: <YY >

NS3-NS4A Other information : Substrate used for fluorescence titration of a protein diploid <YYY>

MOD-RES Name/Symbol: <YYY> (V) Position: <YYY> anthranilyl Derived by Asp Other information: is <YYY>

MOD-RES Name/Symbol: <YYY> (1) Position: <YYY> amino butyric acid (Abu) At position 1 is Xaa <3?7?> Other information:

SITE Name/Symbol: >?1< (7(.......)1) Position: >< peptide peptide bond -C(0)-0- bond Jas Other information: <YYY>

MOD-RES Name/Symbol: <YYY> (9) Position: >< 3-3102 = 9 Other Information: Tag 12 at position <YYY> 1 <.p> Context:

Asp Asp Ile Val Pro Xaa Ala Met Tyr Thr Trp 1 5 10 * Highlight context #: <?70<

VY Length: <?11<

PRT Type: <YYY> Organ: Synthetic Context <713> Characteristics: <YY.> From NS3 protease NS3 Other information: Substrate used for radiometric titration of <YYY> recombinant HCV protease : a <g>

Asp Asp Ile Val Pro Cys Ser Met Sex Tyr Thr Trp 1 : 5 10 Discrimination Context No.: ? <701< 11 Length: SARE:

PRT Type: >< Organ: Industrial Context YY Characteristics: <YY.> 1193 protease NS3 Other information: Tracer used to titrate protease <YYY>

YYVYY

Va

MOD-RES Name/Symbol: <?1< (1) Position: <YYY><> Other information: Asp is processed at position 1 by biotinyl

MOD-RES Name/Symbol: <17 71> (0) Position: <YYY> 1-125 b 01 at position Tyr Other information: Processes <YYY> <> Context: ?

Asp Asp Ile Val Pro Cys Ser Met Ser Tyr Thr Trp 1 5 10 <10?> Context Highlight No.: § + Length: <””11>

PRT Type: ><>713< Member: Industrial context <YY.> Features: <> Other information: Considered as a carbon cleavage product of protease NS3 1153 > Name/Symbol: MOD-RES <> position: (4) <> other information: addresses Tyr at position 4 by 1-125 .> context: ; Ser Met Ser Tyr Thr Trp 1 5 5 Tambez Context No.: <10.< 4 Length: <11< PRT Type: <”717< Sh Member: Synthetic Context <771> Features: chymotrypsin Other information: Synthetic substrate for titration of chymotrypsin <YYY>

MOD-RES Name/Symbol: <Yvy> <> Position: (1) <> A Other Information: Ala at position 1 is processed by succinyl <YYY> Name/Symbol : MOD-RES <YYY> position: (£) <77> Other information: Marks Phe at position ; at the carboxylic terminus para- (pNA) — C-terminal nitro aniline <m. )> Context: #0 Ala Ala Pro Phe 1 YIVYY

Claims (1)

:)[( in the form compound 1 -1 17221 W. 2C xX 2 ZF Q 3 Y Lb = H 0 5 Lm 1 A 2 A 7 1 0 13 M .~ ee R¢ O° Cua v or l CH represents W ¢ from alkoxy «Cy haloalkyl min cycloalkyl »Cy alkyl ¢halo ¢H J—— 5 oy C ° group (Hoa all 82 is Cua (N(RP), or hydroxy «Css cycloalkoxy 1] ¢Cs.6 cycloalkyl from or alkyl L “Cy thioalkyl min haloalkyl” Cy cycloalkyl morph alkyl ¢halo 1 Jou oR? A or Co aryl or Ce aryl “Cy cycloalkyl butt alkoxyalkyl mo cycloalkoxy conjugated alkoxy 9 or unsaturated saturated saturated heterocycle Represents a mixed ring Het Cus © 01 of five ; Six or seven members and with from one to four atoms 0 1 'sulfur oxygen' chosen from 01050860 and heteroatoms VY (R™ mentioned in place of Het or aryl cycloalkyl and bearing VY YYVY Vary «Cy alkoxy «Cs cycloalkyl «Cis alkyl ¢halo group ¢H J— 30 R™ where 4 where each co' (NH-C(O)-NH-R® § NH-C(O) -R* ethane <NO, «Cs cycloalkoxy Vo ¢Cy cycloalkyl atolate of any <H is separately i ¢Cy cycloalkyl from any alkyl (ia R?) where (NH-C(O) )-OR* emfl R* VY or Cg aryl Jas 83 where oNH-RY or a group of the formula NH; chydroxy represents 83 YA -C(0)-OR*?R -C (O)-NHR® ¢-C(0)-R** «heteroaryl «Cj, aryl x ¢Cs cycloalkyl of any alkyl representing R¥ where Y atoms may contain 01 saturated or unsaturated with it from © to an alkylene representing a chain D 71 separately selected from 0; 5; heteroatoms on one to three YY heteroatoms where (N-RY or yr «Cpe alkyl R® Cua «-C(0)-R?J Cy cycloalkyl «Cis alkyl Fg Al 7 affinity § s; mb cycloalkyl Yo D in the carbon chain is reduced to three portable substituents On which atom fal Sy or H represents the said RY 7 Cs alkyl; said substituents are selected separately from the group consisting of Yv and “Cy thioalkyl thio” oxo ¢amino ¢ halo ¢ hydroxy “Cys alkoxy “Cys haloalkyl YA where 8 is chosen from the group consisting of -C(O)-NH-R® with the formula amide representing an amide A Ya ¢C.16 aralkyl uren?; Or aryl or aryl “Cy cycloalkyl” C5 alkyl 7 or pharmaceutically acceptable salt “carboxylic acid ies A” or 79 of it. Pharmaceutically acceptable Lala wa ester 791 mentioned from R moiety R', where the oY moiety of the protector GT 7 selects?- a compound of formula 1 (i) different structures represented by the two structural formulas diastereoisomers of the following two Y stereoisomers: (ii) 5 r YivYy A 8 m ha cm NA R_ 4 0 %, 4 cr D D . (ii) m in form sD in form 0 or an amide adjacent to © 0 is an LSA where 0 is bound to a neighboring oF compound of formula 1 according to claim =F . (ii) represented by the form IN representing W where oY ?- the compound of formula 1 according to claim 0 representing 11 or R® where N(RP), or chloro chydroxy «Cys alkoxy Cis alkyl represents each RY Y ¢ Cis alkyl v chooses from the group that Het or phenyl mt alkoxy mt thioalkyl <H represents R® ¢ consisting of: > =\ _R* N N. = / Dn m vd ¢ 5 S with 22 226 AK N ] 77 I \ J / PS NN N N—N ¢ N R# ¢ f 2 ¢ / 2 12 - 12 0 m d — > ] >> [| 7: NN oN ¢ NN Yyvy Vo RA Je 13 (Je ZF ¢ 0 f 2 where Ve (NH-C(O)-NH- R¥ 4} NH-C(0)-R® {NH-R” “Cis alkyl ) To represent each 1 ¢ C, cycloalkyl or “Crs alkyl 11 each put separately Ja where j. alkyl representing R® Cua “(NH-C(0)-OR*) or Yr .C, alkoxy representing 11 or the RY Cua group of compound of formula 1 according to claim —0 1 Het phenyl alkoxy bit where 277 is of the compound of formula 1 according to claim 1 - 1 choose from the group consisting of: Y —== \ R* N N. > FS ssp + —— RM 'J MN NO : R 7 vd ¢ 5 ¢ 5 2 2 1 —~ CF © 1 Ba La Pe XN N N—N gt NO R* Ff of Je ——RH ¢ Rt where 1 ¢{NH-C(O)-R® sl «(NH-R* in the alkyl L of the cycloalkyl of the first of: R® where each represents A Va 9 or (NH-C(0)-OR™ where WSR is defined in claim 4 . .methoxy A compound representing the Cua of the claim ad I Compound of the form -1/ 1 8- The compound of the form Gay I of the claim No Where 277 represents ethoxy or Het Choose from Y Set consisting of: N N / Dan = a TT : f 5 5 26c 2 ] N—N of 0 2# represents 11182 or R® Cua “NH-C(O)- R” represents Cpgalkyl i.e. cycloalkyl of ; 7 or R** is (NH-C(O)-OR® where Crsalkyl Sra R* 5 “22 is less than H Ar and Cre lac 1 +- the compound of formula 1 according to the element protection 10 where 82 is amide of the form NH-C(O)R™ Y or urea of the form NH-C(0)-NH-R** or carbamate of the form ( NH-C(O)-OR” where 1 represents the alkyl of §Cy6 cycloalkyl 2 Cua “carbamate of urea of claim 9, where 8 represents (Gg] the compound of formula 0 -1 C.s cycloalkyl see alkyl Jay v -1 -1 compound with formula 851 of claim 01 where 83 is carbamate and RP? is cyclobutyl “tert-butyl 7 or cyclopentyl yay . Saturated or an alkylene representing a D chain where 1 of the protectant G51 compounded with formula =) Y 1 heteroatom and may contain a hetero-atom ed 3A to 1 unsaturated thereof from Y Cie alkyl ¢H represents RY one or two separately selected from: ©0;8 or 01-89 where v .N-C,; acyl or optionally a D heterotom containing Cus VY depending on the claim I the compound of formula 1-1 N-C, acyl or NH one chooses from heteroatom 7 mentioned from the heteroatom where the heteroatom (VY) chooses the compound according to claim 0-1 N(Ac) § NH Y mentioned on the © chain © where the chain AY of the claimant Way of the compound contains - 1 .atoms the mentioned VY Y atoms at the heteroatom where the heteroatom (VO) of the compound according to the claim is located 7 -1 mentioned. © chain © of series 01 No. position gaia gall Y mentioned saturated © chain © where chain VY of the claim (Ey compound 11-1 .saturated Y saturated alkylene representing D chain Cua VY compound of formula 1 according to claim 08-1 atoms and optionally containing A to 1 or unsaturated 0 in it from 0 Y S. One choose from © or heteroatom on a different atom v YIVY Maha 1 4- The compound according to the claim Cua A The mentioned 0 chain © contains Y no atoms. 1-01 compound according to claim 8 where the heteroatom is located at position ¥ number 4 of the mentioned D chain D. 1-71-1 Compound 85) of the claim Cua oe the mentioned chain D © bears a substituent of Y 7 at position A where H Cras R* or an alkyl group of 17 1 Compound of claim RN© where RY represents H or .methyl \??'?- Compound 85 of claim (YY where R* represents H 8(S) Me 1 - compound, fad, of claimant Yy in which said D chain D chain is saturated .saturated Y \©?- Gag oS Hall for claimant 8 1 Cus Said chain D© contains one double bond Aad Y between positions 11 and AY 1 1?- the compound of claim (YO) where the double bond of said 7 is positioned opposite trans -71 1 the compound of formula 1 pursuant to claim ? 1 where © is an alkylene chain chain is saturated or unsaturated consisting of all carbon atoms of r number from 1 to 8. Vad Dp where Y series of the Lagi claimant of the formula 74-1 contains atoms. ?- A compound of formula 1 .saturated Y deprecated © chain D, where the chain (Y4) carries the compound of claim 0 -1 .alkyl or alkoxy ¢hydroxy oxo ¢H Ja RY where RY is Substituting from 7 R ligands. The aforementioned H represents RY where 0 is the protection pail Gig The compound =F) \ methyl § H mentioned represents RY where oF) of the protective element Gy "?- The compound (Me \ 10S) or H mentioned represents RY where (TY) of the protectant Gy compound VY 1 has a double bond D where the chain contains 0 YA of the protectant lag; 14 and 1" mentioned between the two positions Y double bond The double bond is Cua » Yo of the claimant (dg 1 a - the compound of formula 1). Mentioned in adjacent position 7 No M” 1 1?- the compound according to the claim (YR) where the aforementioned D chain D chain bears a substituent of “R*Y” where RY represents the alkoxy ¢hydroxy ¢oxo ¢H Yemen or alkyl of .—VA 1 Compound Gig of claimant (FV) where said RY represents H Crealkyl 1 4- Compound (Ey of claimant FA where Said RY represents H, i.e. 1 methyl 6- compound of claim 4 oF, where said RY represents Qa or 4-(-10.1)£—compound of claim 1 of formula )¢ where A is carboxylic acid 1 .y - compound of claim 1 0 where W is IN RC 7 representing a group with the formula -NH-C(O)-NHR “R”-NH-C(0)-OR® 1 where Ce cycloalkyl of D ¢ represents the 41 saturated or unsaturated alkylene chain with 41 of 1 to ° atoms and linked to L in a manner adjacent to A and may contain one heteroatom 1 or two heteroatoms chosen separately from 0<5 or RM Cua “(N-R* represents H 7 or ¢C, .7 acyl JH SR A Tasty substitutes for three independently selected substituents of 9 hydroxy or alkyl of ; 3 A 1 represents carboxylic acid or a pharmaceutically acceptable salt Pharmaceutically acceptable salt or 1 7 pharmacologically acceptable salt. —£F 1 compound of formula [1 pursuant to claim 47; Where A does not represent H R (methoxy R” Y represents Cue alkoxy or Het is chosen from the group consisting of: A YN N N JIN ; 5 3 5 r 26 J ZA ] J > where (NH-C(O)-NH-R* § NH-C(O)-R” (NH-R® U alkyl “T” represents Al4 ¢C, cycloalkyl of Crsalkyl represents 1 of 677 Cua v (Cs cycloalkyl from Ar alkyl where T represents (NH-C(0)-OR™ or “17” represents A 9 and 2277 represents the H or Cpe alkyl group “NH-C(0)-OR¥ in the form carbamate § NH-C(0)-NHR™ in the form urea represents R’ 1 alkyl Jia RY Cua 11 of cycloalkyl of of; b © represents an alkylene chain with 1 atoms and may contain one double bond a between positions 1 1 and 1 Y or 1 1 and Ve ¢ and the chain may contain The aforementioned D Ve on a single 0 hetero-atom chosen separately from © N(Me) «NH ¢S or 'N(Ac) yo and Rs 1 represents Crs alkyl 1 ;- Compound of formula 1 according to the claim? emethoxy JaR? Cua of and “2 Jas ethoxy Y or RA ~r where ¢ YIVY R*® ° NH-C(0)- «NH-C(0)-(C.4 alkyl) <NH-(Cs¢ cycloalky) ¢NH-(C,4 alkyl) alky} ) 1c6)-(0)a {NH-C(O)-NH-(C 4 alkyl) for D represents a saturated chain that is all 1 carbon or contains ada A single double bond in a contiguous position between positions 1” and NE \ 80 — compound of the form: N RZ AC 7 | B 0 z Y N 8 1 COOH 0 R 3 r !6 0 PY {1 6 2 i AN 13 © 12 10 8 v has one stereoisomer at Cua the double bond is recognised. Optional bond ¢ between 1 Y and NY The stereochemistry of a bond ° group R' at position R* 5¢) Y is as follows: > compound number . md double bond Stereochemistry of the 8!RE bond 1" and 109 at position VY ¢1R of Baqem-1 VY 011 position 0 adjacent to ¢phenyl to amide Z N/A 1R position 1 1 ¢ phenyl adjacent to acid and 011 N/A “IR position 0 phenyl-adjacent to MLA Compound with the formula: - 7 1 p N RZ ©: i.e. 9 3 Y 0 N 8 1 coor % Ry | i” 7 6 Cr 9 AH 8. \, d 2 4 Nh R* 3, bond RY recognizes a single Cua RE at a stereoisomer comprising a stereoisomer 7 stereochemistry (VE and 1” OR - 1" 01 H NH-Boc 9 Paraacidic H H Vi Loc (IS or Liacam-17 IR) 0 H NH-acetyl Y.v Paraacidic 1 H VE Loc 5 J IR Not Found 011-91 NH-Boc Yao Contiguous 01-1 Contiguous 1 H Contiguous VE Position (IR Rgm-0 7 H NH-Boc 1 Contiguous 3 OMe Rgm 4 Position “IR sla YE 0 H NH-Boc Bac for L1 acid Yet phenyl OMe adjacent to VE locus (IR sla) EAT H NH-Boc T+ phenyl OMe ste 14 locus IR 0 7 H NH-tBu - 4 tphenyl ~~ OMe rangum 14 15 position gleam) En vw H NH-Boc a for acid phenyl ~~ OMe ragam VE position IR ragam-1 Lar H NH 99 for acid 'H OMe Adjacent to 04 position IR rwag-0 OF H oH nr (one isomer) of phenyl OMe rwagm VE position IR rwagm-01407 HO NE-Boe ne of phenyl OMe rwagm 14 Position IR Not Found H NH-Boc Te of the acid (phenyl OMe fluorescence) VE position (IR contiguous 0 b 10-OH NH-Boc mY (a mixture of the acid) 2 blanks) phenyl OMe 14 position (IR oleae) EAT 10-0 NH-Boc 1" amide for phenyl ~~ OMe VE graphite position (IR not found) H NH- Ac na of the acid iS OMe rgm 1; position (IR rgm-1 0 H mfla 01 and l eee l1 Y.0 : compound of formula 1 1 2 N RZ Q 3 Y 0 N 8 COOH R! y , cut Rr? D one for me where you are n ©» Chemistry stereoisomer includes stereoisomer v JSR? RUDRA seedy for the stereochemistry association: Stereochemistry 0g 2c -D- 8 No. ° D-R! The compound of the tH H bond (1S 4 IR pe NH-Boc 9.1 1s is adjacent to the acid D , . 0 14 yen ,. 10 12 13 12 ¢Ph OMe Rgm © IR 7 9 7 NH-Boc YoY Borg amide L 8 16 1 ¢Ph Mg © 18 Nr NH-B N 7 OMe Mir 00 # A AALS TT un amide for ¢ Ph OMe Rwajam © 18 0 #* NH-Boc Yat . ro N— B 2 14 ¢Ph OMe rogum DIR + , 8 L HO Y.o NN Ph OMe | Contiguous 0 “IR Tr a ts NH-Boc A ~: 10 YYVY von amide for door OMe adjacent D “IR AAD NH-Boc 1 7 2 15 for acid L+- ™ 10 12 14 0 OMe contiguous D «IR ) “NH-Ac YA acid” om, / acid 8 10 BB . Compound with the formula “| $ A \ x Q Y 0 N 8 1 COOH 0 7 lw IX 3 0 0 x, mo allyl bib 3 I 10 13 Xo «R* where the bond of group A is at Position B is “adjacent to the acid” and 7 is defined as follows: VY and 11 optional between double bond ¢ VY and 11 double between positions Aad Xo R* compound number ° VS CH, H 61 adjacent CH, H 7 VS 0 H ay M 0 VS 3 YOR VS 0 V4 M Me N/A 0 H £1 The 1 blabla for 0 not found; Me 0 / 0 does not exist; Me 4 / 0 adjacent; Me 1 0 ¢ / 5 vs. ¢ Me $Y / 5 adjacent ¢ Me ee es sole OD 1 48 - Compound with the formula: 4 apa 0 "0 9 Y : N 8 1 coon 0 a | 34 ' 0 PY 7 3 3 14 12 >< 2 0 Nyt =X 9 13 2 The bond of group R' at position 3" is adjacent to acid 10 and is Xo ese EE TS TR. Bl 'CH, 0 CH, oy CH, CH, CH, Ow 7 ¢ 'NH CH, CH, oO. Y 7 0 A 'N(Me) CH, CH, out {N(CO)Me CH, CH, 0.0 ¢N(CO)Ph CH, CH, 9.1 ¢CH, CH, NH ov CH, CH, N(CO) Me 0vAy : compound of formula 5 -6 1 RA N RZ ~ > 9 Y 0 0 E<A 8 COCH LY 0 HB 0d 10 0 12 14 2 9 11 13 adjacent to acid 40+; And it is 1 at position R and where the group bond is: and with knowledge as follows R* § R22 Complex number 21J 5 0 N(Me), 61 mm N Je IN ¢( CF3) OH TY "J OMe 0117 and orla Y 0 a : Compound with the form —o) 1 p b 0m oy Q Y oO 0 N 8 COOH ST Sp 0 y 7,8 1000 H Mattia 12 7 14 9 R 11 B «R* and be cacid adjacent to the acid AR at position R' where the group bond is v : and 2 defined as follows 1 between Lal and the double bond a the double bond "Xio" and k $ the double bond between Xo number p aa VE and 11 compound B at the two positions {Phenyl adjacent CH= Xo H Yd CH= Xo 0=Xn adjacent right CH, H yl ¢ (for all) Li+- does not exist Hm CH, H .l ¢ (for all) of oO contiguous CH, H 6 for 1 NZ (for all) N contiguous 1m CH , H Vio S (for everyone). CH, H Vay adjacent to Naas core (for all) x no CH H free 8 2 adjacent to N (for all) 7 for +- Ro 5 : CH H 74 2 not found Nae (for all) 1 ]_+- ; CH, H YY not found (for all) N= for u+- ¢ CH, H AR not found 0 (for all) > . 7 for CH, H contiguous +-OFEt (for all) of N 32 : H CH, not present J J (for all) | 4 but CH, H does not exist “-OFEt (for all) ML CH, H contiguous 7 (for all) L and 4” CH, H 71 contiguous 0 (for all) 0 for CH, H mag / wr nd 7 (for all) oN TY Contiguous CH, H YA A (for all) lm 7 Contiguous CH, H 73 (for all) y for 8 Not found CH, H VY. Nae- +! 1 (for all) | N/A CH, H 1 for 3 oS (all) Contiguous A contiguous CH, H 7 CT (for all) 1 2 J N/A CH, H for N (for all) i 8 0 33 : CH H for no Nao not mojo 2 —T T (for all) adjacent to CH, H for (for all) ¢ - + a adjacent to CH, H AR \ (for all) ¢«-CH,-OMe adjacent to CH, H to no (for all) 'Me adjacent to CH, H VYA A , YAY (for all) 8 contiguous CH, H vY4 N ~N —<T (for all) 8 0 3 : CH H YY» Na Unmojo 2 y ! ~T I (for all) Na NH; adjacent CH, H VY — I (for all) 0 adjacent CH; H for (for all) 0 contiguous and CH, H for (for all) lb b contiguous H but 7 / (for all) N—N oF Na contiguous m“j8#8 CH, H yYo : —TI (all) Nae 8 ay contiguous CH, H VY ¢ —T T (all) contiguous CH, H Yep ¢ NZ (all) 2 adjacent CH, H VY A a (all) ¢Ph not present CH, 10-R)yMe 4 (all) 0 CH, 10-(S) Me zx does not exist 'Ph (for all) CH, H VEY 4 dah (for all) adjacent to +- —-o¥ \ Compound of the form: N RZ m 0 to Q Y 08 m 8 the 0 0 Dd > | 8 the oy, 0 R 2 ? NPY, 12 2 14 R* | 1 13 9 v where the 14-8 bond is adjacent to the acid; The double bond is between positions 11 and ; 1 in contiguous mode, and 87 8*2 RES is defined as follows: > complex number “1 8 2 a A Ls p ¢ ¢OEt H n-Pr AY i 0 H Ae Naw Y pe —<T : a 10 Nr H Ae ~~ CL 'r ¢OFt H CY Ad 'OEt H AY Sy ¢OEt H PY Arh p7 Heb 0 l 7 IT Nas Re 8 CL a —T d 0 8 of “ 8 of 8 > AVY TI 7 OF: 8 x AY {7 8 km = SY 7 SP SMS 0 AVY z -+] > wv YYVY <r but LT or \_§s§ © Ir ' bul ah 5 0 a 7 IY 1 bul 7 oT "2 «OF 10-(R) Me CL AYE r t not Add 1 0 27 is a compound with the formula: 2p N 0 Pd J Y 0 o 0 N 8 COOH C | 4 6 ’ 71 14 12 Ha R N 13 d 4c ' r where the 14-8 bond is in an acid-adjacent position; The double bond between 1 at positions 1” and AVE is an adjacent position of 0 and RY “8 and 82 are defined as follows: Complex number R*R32 c a” Qet H CL 9.1 p. HCL 8.7. 0 5 ¢ 4 yla H 47 y 5 ¢ H 4.¢ CL 0 y N—N J 0 l cy 0 rla a o” i H 4.1 oe 2 H Clo | .v 0 ee NH, H Co +A Ls p - u 1.4 Tr Ge - Re H ay ST Cl x R H 41) IY Cl . Ba H 477 No Ke H VY IY LL YYVY A H CL 43 ¢ 8 0 + - H 1 pt 8 N. Os “Oet 10-(R) Me CI 311 p.1 0—A method for inhibiting replication of hepatitis C virus C by exposing virus Y to an inhibiting amount amount of hepatitis C viral 1153 protease 3 and hepatitis C viral 1153 protease from the compound in the form Gag of protective agent 1. 1 00—Jui pharmaceutical composition containing an effective amount against CAV anti-hepatitis © virally effective amount of the compound in formula 1 according to 1 of claim 10 or a therapeutically acceptable salt, i.e. ester ¢ a therapeutically acceptable Ladle thereof; Un saa with carrier medium ° or a pharmaceutically acceptable auxiliary agent. =o 1 Gig pharmaceutical composition of claim 00, also including Y containing immunomodulatory agent —ov 1 Pharmaceutical composition according to claim 00 where Y selects the agent The said immunomodulatory agent is from the category consisting of: .a-, B-, and y-interferons 7 And 1 8©- pharmaceutical composition 4 8) of protectant 00, also includes antiviral agent 1 4- The Ga pharmaceutical composition of protectant OA where he chooses The aforementioned antiviral agent is from the class consisting of: ribavirin .amantadine 9 v -0+1 pharmaceutical composition of claim 00, Y also includes another inhibitor Jafie of protease 1153 of HCV -1 1 pharmaceutical composition pharmaceutical composition per claim 00, also includes a Y inhibitor for other target cells in the HCV life cycle 1-1 method of treatment treating 4 for prevention 48 from cross contamination Contamination with material Y with hepatitis C virus C ASH including the aforementioned sald contact in an amount of 1 effective amount of a compound of formula A according to the claimant 1 - Y 1 A way to treat infection with hepatitis C viral C in mammal a 7 by giving him an amount of Yad against lg) Og liver 0 hepatitis C virally effective amount ¥ of the compound in the form sy] for element 1 Or ¢ therapeutically acceptable salt ade or ester acceptable ade therapeutically acceptable 5 of it. By giving him an amount of 28 YU against the Ca Slr virus YY. pharmaceutical composition anti-hepatitis © virally effective amount 1 00 of claim Ga ¢ 1 5+- pharmaceutical composition According to the claim element dua LY 7 selects the said inhibitor that inhibits a target cell of the phthalate consisting of: v chelicase polymerase And metalloprotease - metalloprotease 1 4- The pharmaceutical composition according to the protection element 0V, where the immunomodulatory agent is -a-interferon Tv 1 pharmaceutical composition according to claim ,*17 also includes 7 antiviral agent. 1 8+- Pharmaceutical composition According to the protection element Cua TY the antiviral agent Y is -ribavirin 1 4- The pharmaceutical composition according to claim 1 also includes Y containing an -antiviral agent -70-1 Pharmaceutical composition according to claim 14, wherein the antiviral agent is -ribavirin 11-1 A way to treat infection with hepatitis C viral C virus Y in humans by administering an amount of Yad against CaS anti-hepatitis C virally effective amount Y from a pharmaceutical composition a pharmaceutical composition ¢ according to claim OV -177 1 Method for the treatment of hepatitis C viral C infection: in humans by administration of YL wd against Ca anti- hepatitis C virally effective amount 3 from Saidapsi formula pharmaceutical composition 3 according to claim 0% 1 “7- A method for treating hepatitis C viral C Y infection in human subjects by giving it fl sa fa 9 against hepatitis C virus C aS ‎anti-hepatitis © virally effective amount v pharmaceutical composition ¢ pursuant to claim 1. 1/4-1 A way to treat hepatitis C viral C 7 infection in humans by administering an effective amount against hepatitis C virus © anti-hepatitis C virally effective amount v pharmaceutical composition ¢ 5 18 of protectant AY Vo 1 A method for treating hepatitis © viral C 0 infection in humans by administration of virally effective amount 7 © 010-6602005 Senior Installation Sidon_C AN According to claim, pharmaceutical composition ¢ hepatitis C viral C method for treating hepatitis C virus infection ‎ anti-hepatitis C virally effective amount 7 YIVY YYY AY According to claim pharmaceutical composition ¢ hepatitis C viral C /the/a- a way to treat hepatitis virus infection 1 0 aS lethal virus Aa 9 Fact I A 3a human administration ¥ from a pharmaceutical composition anti-hepatitis © virally effective amount and IA according to the claim pharmaceutical composition ¢ hepatitis C viral C dalled method 1-78 by administering an effective dose against a virus Hepatitis A human in humans 4 Pharmacist Sy 5 elderly anti-hepatitis C virally effective amount 7 8 According to claim pharmaceutical composition ¢ hepatitis C viral © A method for treating hepatitis C virus infection -4 1 CaS against Yad hepatitis virus by giving it a human dose of Y from a virally effective anti-hepatitis © virally effective amount of Yo according to the claim pharmaceutical composition $ ben alkyl representing R™ -NH-C(0)-OR” is the B® compound of the claim ££ , where =A. 1 or carbon atoms 1 containing carbon representing a fully saturated chain D mm©; cycloalkyl or 1 at a single cis double bond in adjacent position double bond containing a double bond v represents R? 561 E VY Positions ¢ 24 Ne RF : > l 'NH-C(0)-(C,.¢ alkyl) {NH-(Cs.s cycloalkyl) {NH-(Cy.4 alkyl) Jia) where For 1 a alkyl) v b0)-0-(111-0)0; A .NH-C(0)-NH-(C,., alkyl) =A) 1 is a compound with the formula: N RZ 0 Pe TF Q Y : N 5 COOH 0 7 A x > | v 0 8 nb 14 2 12 ps” 7 R N TI 13 v * where the !14-8 bond is in an acid-adjacent position; The double bond between positions 1” and VE is in an adjacent position, and RZ GRR? is defined as toh -their name is a toothy humus<<-<a dam mm doss << ...<> tooth >> A >< > with >> na »that you need a brother>>t atat: naa ><<>>t: make a <<tat << at a ha. Compound number:R%:R? ‘R* _- blood N 8 H Cl. —~_T 0' N. 8 H CL AY pe 4 ]> Na NOY 1 Cl. AYY Yyvy Thap “H Cl. AYA <I p z . 47 for claimant Ty 708 composite 87 0 ET for claimant Gi, 705 composite AY .47 for claimant Ga, YE composite -84 0 . 46 for claimant Gi, 717 compound -+* 0 A for claimant Gy £0A compound -8“ 0 . 49 for Gay 00A Composite Claim -87 0 00 for Composite Gig 100 Claim “AA Ov for Gay 107 Composite Claim - + 0 0 OY for Composite Ga, 717 Claim -40 0 0) for compound 707 according to claim —4) 0 0) for claim iy Ved compound -47 0 YYo 0) of claim Ta, 04 compound 4 1 OY) of claim lag 711 component 4-1 0) of claim la, VIA compound 5-1 0) of claim Wa, 770 Composite 17-1 0) For Gay VFN Composite 7-1 0) Gag VFA Component 4-1 OF For Component Gy Av) Composite-4 1 OF Compound 8094 According to Claim - 0s 1 OF According to Claim 801 Composite -011 1 OF Compound Gag 8011 Compound =) oY 1 OF Compound Gag AVY Compound - 0" \ OF for Gag AYE - 4 Composite Claimant 1 OY for Composite lag AYA Claimant - 1 0 A 1 7 - Compound AYE according to claim OF-01 1 Compound Gig 871 of claim OF-08 1 Compound Gag AYY of claim OF 1 4 - Compound AYY according to claim OY YY 1 Compound 877 Gig of claim AY -11 -1 Compound 904 Gag of claim OF Y 1 )= Compound W444 of claim Protection OF -1 ““ 1 Compound Tay 904 of claim OF 1 64 - Compound 991 according to claim OF Vo 1 Pharmaceutical composition comprising an amount Jad against y anti-hepatitis C virally effective amount of compound r in form 1 Gay of any of the 87 protections to OVE or a therapeutically acceptable salt ¢ OR »1 Lac therapeutically acceptable ° from it; mixed with a carrier medium Cala or a pharmaceutically acceptable auxiliary agent 1. 1 - The pharmaceutical composition according to claim 119, Y also includes an immunomodulatory agent -immunomodulatory agent -11117 1 The pharmaceutical composition according to claim 111, where v Selects the mentioned immunomodulatory agent from the class that consists of 0-,f3-, and y-interferons : je 1 1-8 1 pharmaceutical composition according to claim 111, where Jalal (Y) is the immunomodulatory agent mentioned is of -a-interferon 1 48 - pharmaceutical composition according to claim 111 0 also includes -antiviral agent -1 0 1 pharmaceutical composition according to For claim IV where Y is the antiviral agent is (11027110. -111 1 Gg pharmaceutical composition of claimant 1117, 1 also includes an antiviral agent -antiviral agent -17 1 pharmaceutical composition according to claim IY where 1 said antiviral agent is ribavirin YY \ = Gag pharmaceutical composition of element Protection VIA v also includes -antiviral agent YYVYY YYA NYY pharmaceutical composition 1- 1 ribavirin is antiviral agent Slug pall where antiviral agent is v 110 pharmaceutical composition “xX all - 15© 1 1153 another protease inhibitor also includes a HCV Y repressor for 3 includes 1 V0 according to claim pharmaceutical composition - 4 1 other in the life cycle targets of cells Targeted inhibitor hfe also on Y HCV life cycle v where NYT according to the protective element pharmaceutical composition 171-1 from the category mentioned target that inhibits the target cell inhibitor selects cleft 4 and a metalloprotease helicase, which consists of: helicase 7.metalloprotease ¢ a compound with the following formula (1): YA 1 (0) x 0 v N Ro a "TX PG 0 7 a r where X ¢ represents PG or (R*° each PG individually represents ¢protecting group 48 y SR 1 as defined in claim 1; A' 77 represents carboxylic acid a buffer; A and fae Juan equals ? with 1 is a compound of the formula (—) : X as Ra N oH + - od \ ° ¢ a (c) (b) X oo” CLs : / PG 6 7 (1) Cua is 3A GX PG as defined in AYA Claim A Yr. A compound of formula (d) is: -10” \ (+) * y Y a © A Cua 1 or 2; PG represents X 1 ¢protecting group and protective ac represents the PG group 1°; defined in protective element WS SR? stop; carboxylic acid represents A' 7 Y equals Taxe and « represents A The process includes Cus VY according to claim (J) a process for the preparation of a compound of formula 1-11 pyrrolidine on The aforementioned ring PG protecting group, the protective group Y part in the compound with formula (A): 1 a X o” (Ls um and PG ° A (7) ¢ X y) (Ls . ° A, { (>) \ where ° (R? or PG is X 1 ¢protecting group separately represents a protective group PG each y AY. It is as noted in the claim © 3A’ (R? A) A compound having the following formula (e): TY 1 —) ( ox 0 . 0 0 Y “rp Ry R Iq YYVYY v where X 1 represents PG or (R® PG ° represents protective group ¢protecting group WS GSR 1 defined in claim 1; A' v represents carboxylic acid is temporary; A is as noted in claim 1; 9 “ represents a number equal to zero or 9; and: 1 0) when “ Tare is equal to zero then '0 represents a saturated alkylene chain It has 11 © atoms that optionally contain heteroatoms ranging from one to three VYs, each choosing separately from: ©0; 8; or “018” or (Y)1 when Tae represents n is equal to ?, then DY represents an alkylene chain saturated with ?Ve atoms optionally containing one to three heteroatoms each choosing separately from: SO or IN- RY to and be “© as enclosed in claim 1. Adee —VFF 0 for the preparation of a compound of formula (e) Gy of claim TY wherein said process Y involves reacting a compound of formula (d) with Compound with the formula (and): YYIVYY yyy X y0 0 8 A + ~N — 3 [ 7 (d 0 r X y 8 a cr for d 0 Rr? : (=) AY as defined in the claim D' where “d is a cov y p and 3 is a compound with the following formula (g): 1 - 1 Rr? 4 7 ' 8 A Y 0 0 7 a <> bo (g) where r;1 be and 3 as defined in the claim YyVY A’ ° is carboxylic acid buffer; 1 « represents a number equal to zero or 7; And 7 0) When n is a number equal to zero, D7 has an alkylene chain saturated with © A atoms that optionally contains heteroatoms numbering from one to 9 three of which JS chooses sharpness from: 0 8; or maybe or (Y) 01 when Jia « is a number equal to 7; The 0' represents a saturated alkylene chain with 11 “atoms” that optionally contain heteroatoms ranging from one to three VYs, each individually selected from: 0« 8 or (N-RY) 9" and WSRY is roomed in the claim .1 ATE of the protective film Udy is a process for preparing a compound with the formula (g) IYO 01, where the mentioned process includes closing the ring of the compound with the formula (them) auld by reacting the compound with the formula (e) in the presence of a catalyst 1 to obtain< transition metal-based catalyst $ with the formula (g*): ° Yye X 0” 2 8 0 They are 0 . Na 0 R yda \ (=) i « o” N Rw A pay 2 “py m amag (g) a where ¢R? Or, PG Jia X A ¢protecting group 48) 5 represents the PG 4 group (Ye Gravette in the protective element LSA "sn the R® (R? P), the protective group is removed from the compound of the formula PG Je X and when 11 : (0) to obtain a compound of the formula R*-OH reacts further with a compound of the formula ( ¢ J) VY A x 6 Row 0 Ri—oH_ the 6 R f > h (2) , 1 Qo” 3 \Y N | b 0 : 0 8 "ny h (3) AYE as defined in claim A' D and D' (R® (R? where Ve is a compound with the following (0) formula: VTA 1 R2 o “Char Ale : 0 0 ” ry h (2) Cus Y 1 customarily in the claim WSR? 3 R? It is 3 times; carboxylic acid represents A 1° YYy and ¢Y a number equal to zero or Jaan 1 saturated with alkylene atoms equal to zero then 07 represents a fade n chain when (V) represents 7 optionally containing 85 hetero-atoms with a number of One to A or NR three to choose from: 0« 8; or 4 N containing NY saturated alkylene series Jha DD ?; Then « Je when ( Y ) 01 optionally on hetero-atoms 35 ranging in number from one to three 11 select each separately from: 0 8; or 4m; VY A as chambered in the claim RH and be VY wherein the process includes 6, 1 of the claim Gy to prepare a compound of the formula (h) lee -117 1 'hydrogenation mentioned exposing the compound with the formula (g) to the hydrogenation process y R2 o N BR. a ia DA 0 > 0 RS oy oe h v (3) 0” N 8 A ° i o 3 X © h (2) ATT as defined in element Protection A gn «D' (RRP where ¢ -1 YA 1 is a compound with the following formula (i: / Q COA % 8 N \ 0 a Ba NHCbz : (+) Cua 01 ¢ is 2 and T as they are known in element Protection 1; Jar m ° is a number from 1 to co 4 « represents a number from 1 to 0 for A' carboxylic acid represents a buffer ¢ A and Cbz .benzyloxycarbonyl 1 4- represents a compound with the following formula (k): 2 and % A - CY : i 5 — 0 “ETN Enemy R (¢) a v where ¢ is a scalar as defined in claim 1 Sam 5 is a number from 1 to 0 Jan 1 is a number from 1 to 0 A' 7 is a buffer carboxylic acid; A and Cbz represent benzyloxycarbonyl. Then subjecting it to an oxidation process, rR?d . . 3 Al / 0) boron addition NR SAS SS . o== () oxidation 0 7 R Fm ym (0) / 9 Cx : N 1 0 0 mm” as 8 (4) : 0 where yin is “<n” m R® has Cbzg as defined in the protection element Ara Ye. The following: (J) is a compound of the formula VED 1 R32 q, % A GAY 0 1 7 Y - PT SN R i (J) where v;1 by convention in the WER claim is 87 and ¢ 0 to 1 represents a number from m ° 0 to 1 representing a number from © 1 ¢ 3 3a carboxylic acid represents A's 7 wherein process , 511 of claim (aa (J)) includes a process for preparing a compound of formula 1-31 1 in the presence of said hydrogenation subjecting the compound of formula (k) to a hydrogenation process Y racid acid v 2c / C A' 9 hydrogenation of acid $ ; | CA ——————— 0 0 0= “ETN rR NHCbz ¢ (a) . / Q A ” 0 0 0 ETT SS R b (J) 2 where 1 is R? and no d is d and WA is sweated in the protective element AREA of 5 Cbz represents .benzyloxycarbonyl -04 Y 1 is a way to prevent infection hepatitis C viral infection C ASH Y in mammal administration includes administration of hepatitis © virally effective amount C 1 of compound as Formula 1 of claim 1¢, or salt Therapeutically acceptable salt dle or ester accepted ade therapeutically acceptable ° from it VEE 1 A method for treating hepatitis C viral infection C Y in mammal disease includes giving it Yad against the A virus YEY (38g ] from a combination with the compound in the form hepatitis © virally effective amount liver 7 ester any therapeutically acceptable salt Ladle or acceptable salt 1 ¢ for one additional agent and additive ,4% therapeutically acceptable Ladle. Other in the life cycle target of a target cell Inhibitor and activator A mentioned at least one before, or additional agent Sua HCV a additive agent given to protective agent Gig 1 compounded in form administration in conjunction with, Or after giving 01 therapeutically acceptable ester or therapeutically acceptable salt Ladle or therapeutically acceptable salt 1, 1 of it. VY hepatitis C viral infection C 1- EO 1 C includes administration of an effective amount against mammalian hepatitis virus in hands Y in combination with the compound in formula 1 according to an effective amount hepatitis C virally Y Ladle ester or therapeutically acceptable salt Ladle Or an acceptable salt, 1 Protection ¢ At least one of them chooses an additional agent from it, and a therapeutically acceptable addition agent ° Antiviral agent, immunomodulatory agent from factor 4 inhibitor L 17, and cut 1153 another protease inhibitor antiviral agent L where another 1107 life cycle additive agent is administered in the life cycle target of a target cell A at least one mentioned before, in conjunction with, or After additional agent 9 ade or acceptable salt, 1, the compound of formula 1 5 88 (to the protective agent 0 ye) is administered thereof. therapeutically acceptable Ladle ester or therapeutically acceptable salt 1