TW201249786A

TW201249786A - Antibacterial agents

Info

Publication number: TW201249786A
Application number: TW100140449A
Authority: TW
Inventors: Ramesh Kasar; Martin Sheringham Linsell; James Bradley Aggen; Qing Jane Lu; Dan Wang; Tim Church; Heinz E Moser; Phillip A Patten
Original assignee: Achaogen Inc
Priority date: 2010-11-10
Filing date: 2011-11-04
Publication date: 2012-12-16
Also published as: AR083760A1; SG190243A1; BR112013011693A2; EA201390626A1; JP2014501716A; AU2011367819A1; IL226270A0; EP2638006A1; MX2013005200A; WO2012154204A1; KR20140023869A; CA2817211A1; CN103298780A

Abstract

Antibacterial compounds of Formula I are provided: as well as stereoisomers and pharmaceutically acceptable salts thereof; pharmaceutical compositions comprising such compounds; methods of treating bacterial infections by the administration of such compounds; use of such compounds in the treatment of bacterial infections and processes for the preparation of such compounds.

Description

201249786 六、發明說明：【發明所屬之技術領域】本發明大體而言係關於治療由革蘭氏陰性細菌（gramnegative bacteria)所引起之感染。更特定言之，本文所述之本發明係關於藉由抑制UDP-3-0-(R-3-羥基癸醯基）_N-乙醯葡萄糖胺去乙醯基酶（LpxC)之活性來治療革蘭氏陰性感染。本發明提供LpxC之小分子抑制劑、含有該等抑制劑之醫藥調配物、用該等醫藥調配物治療患者之方法及製備該等醫藥調配物及抑制劑之方法。本文所述之本發明係關於藉由單獨投與或與第二抗菌劑組合投與能抑制UDP-3-O-(R-3-羥基癸醯基）-N-乙醯葡萄糖胺去乙醯基酶（LpxC)之活性之化合物來治療革蘭氏陰性感染。相關申請案之交叉引用本申請案根據35U.S.C.§119(e)主張2010年11月1〇日申請之美國臨時專利申請案第61/412,311號之權益，該申請案以全文引用的方式併入本文中。政府利益聲明本發明根據美國國防部（Untied States Department of201249786 VI. Description of the Invention: TECHNICAL FIELD OF THE INVENTION The present invention relates generally to the treatment of infections caused by gramnegative bacteria. More specifically, the invention described herein relates to the treatment by inhibiting the activity of UDP-3-0-(R-3-hydroxyindolyl)-N-acetylglucosamine de-acetylase (LpxC). Gram-negative infection. The present invention provides small molecule inhibitors of LpxC, pharmaceutical formulations containing such inhibitors, methods of treating such patients with such pharmaceutical formulations, and methods of preparing such pharmaceutical formulations and inhibitors. The invention described herein relates to the inhibition of UDP-3-O-(R-3-hydroxyindolyl)-N-ethylglucosamine by decoupling by administration alone or in combination with a second antibacterial agent. A compound that is active in the enzyme (LpxC) to treat Gram-negative infections. CROSS-REFERENCE TO RELATED APPLICATIONS This application is hereby incorporated by reference in its entirety to the entire entire entire entire entire entire entire entire entire entire entire entire entire disclosure Into this article. Statement of Government Interests This invention is based on the United States Department of Defense (Untied States Department of

Defense)授予之合同HDTRA1-07-C-0079在政府支持下進行。政府對本發明享有某些權利。【先前技術】在過去幾十年中，抗微生物抗性之頻率及其與嚴重傳染病之關聯已以驚人速率增長。抗菌抗性之問題伴隨存在對多種抗菌劑有抗性之菌株。因此，對新穎抗菌劑，尤其是 159849.doc 201249786 具有新穎作用機制之抗菌劑存在需要。先前未開發但高度保守之目標LpxC提供一個新的開發廣效抗菌小分子之機會，該等廣效抗菌小分子包含新一類應遭遇極小（若存在）天然存在之目標相關抗性之活性殺菌化學實體。LpxC(酶尿普二填酸-3-0-(R-經基癸酿基）-N-乙醯葡萄糖胺去乙醢基酶）存在於所有相關革蘭氏陰性細菌物種中且與外膜生物合成中之第一關鍵步驟有關。因此，LpxC為生存所必需且為革籣氏陰性細菌物種中之抗細菌活性呈現理想目標。研究者已鑑別出一些靶向脂質A生物合成之具有抗菌活性之化合物。舉例而言’ Jackman等人（J· Biol. Chem.， 2000，275(15)，11002-11009) ; Wyckoff 等人（Trends in Microbiology，1998, 6(4)，154-159);美國專利申請公開案第2001/0〇53555號（公開於2001年12月20日，對應於公開於 1998年5月7日之國際PCT公開案第WO 98/18754號）；國際 PCT公開案第WO 00/61134號（公開於2000年1〇月19曰）；美國專利申凊公開案第2004/0229955號（公開於2004年11月18 日），及國際PCT公開案第W0 2008/1 54642號（公開於2〇〇8 年12月1 8日）均揭示具有抗菌性抗LpXc活性之化合物。此等LpxC抑制劑之商業性開發由於濃度處於或接近抗菌活性所需之濃度的此等化合物在哺乳動物中之毒性而複雜化。與具有抗LpxC活性之其他緊密相關化合物相比，本文所呈現之化合物的耐受性明顯更佳。儘管在該領域已有進步，但仍需要具有活性以作為針對革蘭氏陰性細菌之殺菌劑且具有可接受之毒性/耐受性概 159849.doc 201249786 況之LpxC抑制劑。因此，本發明之一個目標為提供用於製備無毒抗菌劑及能夠抑制革蘭氏陰性細菌感染之其他藥物的化合物及該等化合物之組合。本發明之另一目標為提供抗菌劑與LpxC抑制劑之協同組合，其具有固有抗菌性質以及改良革蘭氏陰性細菌之外膜對其他抗菌劑之滲透性的能力。使用藥物之協同組合可具有許多優於習知單一化合物化學療法之優勢，包括較低使用劑量或較短治療時間使得藥物副作用降低、較快治癒感染使得住院期縮短所防治之病原體的範圍增大及對抗生素產生抗性之發生率降低。【發明内容】本發明提供新穎化合物、包括該等化合物之醫藥調配物、抑制UDP-3-〇-(R-3_經基癸醯基）_N_乙醯葡萄糖胺去乙醯基酶（LpxC)之方法及治療革蘭氏陰性細菌感染之方法。在一態樣中’本發明提供化合物：The contract awarded by Defense) HDTRA1-07-C-0079 was carried out with government support. The government has certain rights in the invention. [Prior Art] The frequency of antimicrobial resistance and its association with severe infectious diseases has increased at an alarming rate over the past few decades. The problem of antimicrobial resistance is accompanied by the presence of strains that are resistant to a variety of antimicrobial agents. Therefore, there is a need for novel antibacterial agents, particularly those having a novel mechanism of action, 159849.doc 201249786. The previously undeveloped but highly conserved target LpxC offers a new opportunity to develop broad-spectrum antibacterial small molecules containing a new class of active bactericidal chemistry that should suffer from minimal (if any) naturally occurring target-related resistance. entity. LpxC (enzyme urinary dihydrogenate-3-0-(R-ionyl)-N-acetylglucosamine deacetylase) is present in all relevant Gram-negative bacterial species and with the outer membrane The first key step in biosynthesis is related. Therefore, LpxC is essential for survival and is an ideal target for antibacterial activity in Gram-negative bacterial species. Researchers have identified a number of compounds that have antibacterial activity that target lipid A biosynthesis. For example, 'Jackman et al. (J. Biol. Chem., 2000, 275(15), 11002-11009); Wyckoff et al. (Trends in Microbiology, 1998, 6(4), 154-159); US Patent Application Publication No. 2001/0〇53555 (published on December 20, 2001, corresponding to International PCT Publication No. WO 98/18754, filed on May 7, 1998); International PCT Publication No. WO 00/ No. 61134 (published in January 19, 2000); US Patent Application Publication No. 2004/0229955 (published on November 18, 2004), and International PCT Publication No. WO 2008/1 54642 (public) Compounds having antibacterial anti-LpXc activity were disclosed on December 18, 2008. Commercial development of such LpxC inhibitors is complicated by the toxicity of such compounds in mammals at concentrations at or near the concentration required for antimicrobial activity. The compounds presented herein are significantly more tolerant than other closely related compounds having anti-LpxC activity. Despite advances in this field, there is still a need for LpxC inhibitors that are active as fungicides against Gram-negative bacteria and have acceptable toxicity/tolerability. 159849.doc 201249786. Accordingly, it is an object of the present invention to provide compounds and combinations of such compounds for the preparation of non-toxic antibacterial agents and other drugs which inhibit Gram-negative bacterial infection. Another object of the present invention is to provide a synergistic combination of an antimicrobial agent and an LpxC inhibitor which has inherent antibacterial properties and the ability to improve the permeability of membranes other than Gram-negative bacteria to other antimicrobial agents. The synergistic combination of drugs can have many advantages over conventional single compound chemotherapy, including lower doses or shorter treatment times, resulting in reduced side effects of the drug, faster healing of the infection, and an increase in the range of pathogens that are prevented by the shortened hospital stay. And the incidence of resistance to antibiotics is reduced. SUMMARY OF THE INVENTION The present invention provides novel compounds, pharmaceutical formulations including the same, and inhibition of UDP-3-oxime-(R-3_transylhydrazino)_N_acetylglucosamine deacetylase (LpxC) And methods of treating Gram-negative bacterial infections. In one aspect, the invention provides a compound:

及其立體異構體及醫藥學上可接受之鹽，#中A為經取代之Cs-C6環烷基，其中至少一個取代基為一級醇；b 不存在或為_CH=CH-、_C=C-或未經取代之苯基；c為 …、_C5C-或未經取代之苯基，其中若B為-CH=CH-，則c不會亦為_CH=CH_ ; …及尺3係獨立地選自氫及經取代或未經取代之Ci_C3烷基，或R〗及R2連同其所連接 159849.doc 201249786 之碳原子-起形成未經取代之以環烷基，或。及…連同其所連接之碳原子及Q一起形成具有5至8個環原子之經取代或未經取代之雜環，其中該雜環之⑴個環原子係選自N、〇及S，且Q為〇或抓，其中R為氫或未經取代之 C3烷基》在另-態樣中，本發明提供一種包含式工化合物或其立體異構體或醫藥學上可接受之鹽，及醫藥學上可接受之載劑或稀釋劑的醫藥組合物。在另一態樣中，本發明提供一種包含有效量之式ι之抗菌化合物或其立體異構體或醫藥學上可接受之鹽，及醫藥學上可接受之載劑或稀釋劑的醫藥組合物^ 、〜、樣中本發明提供一種抑制革帛&陰性細菌中去乙醯基酶，藉此影響細菌生長之方法，其包含該抑，之患者投與式kLpxC抑制化合物或其立體異構體; 醫藥學上可接受之鹽。在另：態樣中’本發明提供一種抑制革蘭氏陰性細菌中 LPXC’藉此調整細菌感染之毒性之方法，其包含向需要該。者扠與式I之LpxC抑制化合物或立醫藥學上可接受之鹽。戈在另、t樣中’本發明提供—種治療患有細菌感染之個人的方法’其包含向有需要個體投與抗菌有效量之式I化 ::或其立體異構體或醫藥學上可接受之鹽。在治療方法疋實施例中’細菌感染為革蘭氏陰性細菌感染。個該實施例十’細菌為綠膿假單胞菌(― 159849.doc 201249786 aerwgiwoja)、伯克霍爾德氏菌屬（5wrA:/zo/i/eha)(例如；f ,爹伯克霍爾德菌{Burkholderia cepacia)) '腸桿菌科 (五wierokcieriaceae)、弗朗西絲菌屬(例如土拉熱弗朗西絲菌{Franciscella tularensisY) '妙售氏議屬（<Serraiia)、變形桿菌屬（/VoieMS)、克雷伯氏菌屬 (A7eZ^ie//<2)、腸桿菌屬（五wierofeacier)、檸檬酸桿菌屬 (OMacier)、沙門氏菌（iSa/wc>;ie//a)、普羅威登斯菌屬 (jProWi^«ci<3)、耶氏桿菌屬（FemWa)(例如葳疫#戌#磨 (TierizWa 、摩根氏菌屬（Morga«e//a)或大腸桿菌 co")。在一特.定實施例中，細菌為綠膿假單胞菌、伯克霍爾德氏菌屬、弗朗西絲菌屬、腸桿菌屬、耶氏桿菌屬或大腸桿菌。在一個該實施例中，細菌為綠膿假單胞菌。在另一該實施例中，細菌為大腸桿菌。在另一實施例中，知魯％嗜麥芽窄食單胞菌{Stenotrophomonas maltophila、、木糖氧化產驗菌[Alcaligenes xylosoxidans)、啥▲桿菌屬（Haemophilus)、奈瑟菌屬{Neisseria species)、西地西菌屬（Cedecea)或愛德華氏菌屬（Edwardsiella ipecies)。在另一特定實施例中，個體為哺乳動物，且在某些實施例中為人類。本發明之一態樣提供包含LpxC之抑制劑及第二抗菌劑之醫藥組合物。在一實施例中，該第二抗菌劑係選自由以下組成之群：萬古黴素（vancomycin)、利奈β坐胺（linezolid)、阿奇徽素（azithromycin)、亞胺培南（imipenem)、替考拉寧 (teicoplanin)、達托黴素（daptomycin)、克林達黴素And its stereoisomers and pharmaceutically acceptable salts, wherein A is a substituted Cs-C6 cycloalkyl group, at least one of which is a primary alcohol; b is absent or is _CH=CH-, _C =C- or unsubstituted phenyl; c is ..., _C5C- or unsubstituted phenyl, wherein if B is -CH=CH-, then c will not also be _CH=CH_; Is independently selected from hydrogen and substituted or unsubstituted Ci_C3 alkyl, or R and R2 together with the carbon atom to which they are attached, 159849.doc 201249786, form an unsubstituted cycloalkyl group, or. And ... together with the carbon atom to which it is attached and Q form a substituted or unsubstituted heterocyclic ring having 5 to 8 ring atoms, wherein the (1) ring atom of the hetero ring is selected from N, fluorene and S, and Q is oxime or smear, wherein R is hydrogen or unsubstituted C3 alkyl group. In another aspect, the invention provides a compound comprising a formula, or a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and A pharmaceutical composition of a pharmaceutically acceptable carrier or diluent. In another aspect, the invention provides a pharmaceutical combination comprising an effective amount of an antibacterial compound of the formula i or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent The present invention provides a method for inhibiting deacetylase in a sputum & negative bacterium, thereby affecting the growth of bacteria, which comprises the inhibition of a compound of the formula kLpxC or its stereoisolation Construct; a pharmaceutically acceptable salt. In another aspect, the invention provides a method of inhibiting the toxicity of a bacterial infection by a LPXC' in a Gram-negative bacterium, which comprises the need for this. An LpxC inhibiting compound of formula I or a pharmaceutically acceptable salt. In the alternative, the present invention provides a method for treating an individual having a bacterial infection, which comprises administering an antibacterial effective amount to an individual in need thereof: or a stereoisomer or medicinal Acceptable salt. In the treatment method 疋example, the bacterial infection is a Gram-negative bacterial infection. The tenth example of this example is Pseudomonas aeruginosa (― 159849.doc 201249786 aerwgiwoja), Burkholderia (5wrA:/zo/i/eha) (eg; f, 爹Berke {Burkholderia cepacia)) Enterobacteriaceae (five wierokcieriaceae), Francis (for example, Franciscella tularensis Y) (Serraiia), Proteus (/VoieMS) , Klebsiella (A7eZ^ie//<2), Enterobacter (five wierofeacier), Citrobacter (OMacier), Salmonella (iSa/wc>; ie//a), Providencia Genus (jProWi^«ci<3), FemWa (eg plague #戌#磨(TierizWa, Morgena (Morga«e//a) or E. coli co") In one embodiment, the bacterium is Pseudomonas aeruginosa, Burkholderia, Frances, Enterobacter, Yarrowia or Escherichia coli. In one such embodiment, the bacterium Pseudomonas aeruginosa. In another such embodiment, the bacterium is Escherichia coli. In another embodiment, the genus Stenotrophomonas maltophilia {Stenotrop Homonas maltophila, Alcaligenes xylosoxidans, Haemophilus, Neisseria species, Cedecea or Edwards iella ipecies. In another specific embodiment, the individual is a mammal, and in some embodiments a human. One aspect of the present invention provides a pharmaceutical composition comprising an inhibitor of LpxC and a second antibacterial agent. In one embodiment, the second antimicrobial agent is selected from the group consisting of vancomycin, linezolid, azithromycin, imipenem, Teicoplanin, daptomycin, clindamycin

159849.doc S 201249786 (clindamycin)、利福平（rifampin)、頭孢噻肟（cefotaxime)、慶大黴素（gentamicin)、新生黴素（novobiocin)及特拉萬星 (telavancin)。在一個該實施例中，第二抗菌劑為萬古黴素或利福平。在另一實施例中，LpxC抑制劑為式I化合物：159849.doc S 201249786 (clindamycin), rifampin, cefotaxime, gentamicin, novobiocin, and telavancin. In one such embodiment, the second antimicrobial agent is vancomycin or rifampicin. In another embodiment, the LpxC inhibitor is a compound of formula I:

或其立體異構體或醫藥學上可接受之鹽，其中A為經取代Or a stereoisomer or pharmaceutically acceptable salt thereof, wherein A is substituted

之C^C：6環烷基，其中至少一個取代基為Ci_C3一級醇；B 不存在，或為-CH=CH-、-C^C-或未經取代之苯基；c為 CH CH-、-C=C-或未經取代之苯基，其中若6為_ch=ch_ ，則c不會亦為_CH=CH_ ; R1、…及尺3係獨立地選自氫及經取代或未經取代之Cl_C3烷基，或Rl&R2連同其所連接之碳原子一起形成未經取代之^彳6環烷基，或r2及r3連同其所連接之碳原子一起形成具有5至8個環原子之經取代或未經取代之雜環，其中該雜環之⑴個環原子係選自N、〇及S;且㈣⑽似，其中汉為氫或未經取代 c3烷基。發月之另態樣提供治療患有革蘭氏陰性細菌感染之患者，方法，纟包含共同投與協同量’例如活體内協同量 ::菌劑及式I之LpxC抑制劑。在一實施例中，該抗菌劑去、、自由以下組成之群：萬古黴素、利奈唾胺、阿奇黴、胺。南替考拉甯、達托黴素、克林達黴素、利福 159849.doc 201249786 平、頭孢噻肟、慶大黴素、新生黴素及特拉萬星。在一個該實施例中，抗菌劑為萬古黴素或利福平。參考以下實施方式時將顯而易知本發明之此等及其他態【實施方式】本發明提供新穎化合物、抑制革蘭氏陰性細菌中LpxC之方法及治療細菌感染之新穎方法。本文所提供之化合物可調配成適用於本發明方法之醫藥調配物及藥劑。本發明亦提供化合物在製備藥劑及醫藥調配物中之用途、化合物在抑制LpxC中之用途及化合物在治療個體細菌感染中之用途。本發明進一步提供治療革蘭氏陰性感染之組合物及方法，其係藉由單獨投與能夠抑制UDp_3_〇_(R_3_羥基癸醯基）-Ν-乙醯葡萄糖胺去乙醯基酶（LpxC)之活性的化合物或與投與第一抗菌化合物組合來實現。 A.定義在整個本申請案中使用以下縮寫及定義：「LpxC」為表示UDP_3_〇_(R_3_羥基癸醯基）_N乙醯葡萄糖胺去乙醯基酶之縮寫。除非另外指出，否則如本文所用，以下定義應適用。「燒基」係指具有1至10個碳原子且較佳1至6個碳原子之單價飽和脂族烴基。此術語包括例如直鏈及分支鏈烴基，諸如甲基（CH3·)、乙基(ch3ch2-)、正丙基(CH3CH2CH2-)、異丙基（(CH3)2CH-)、正丁基（CH3CH2CH2CH2_)、異丁基 ((CHACHCH2·)、第二丁基（(CH3)(CH3CH2)CH_)、第三丁 159849.doc 201249786 基（(CH3)3C-)、正戊基（ch3ch2ch2ch2ch2-)及新戊基 ((CH3)3CCH2-)。「烧氧基」係指基團-Ο -烧基，其中烧基係如本文所定義。烷氧基包括曱氧基、乙氧基、正丙氧基、異丙氧基、正丁氧基、第三丁氧基、第二丁氧基、正戊氧基及其類似基團。「胺基」係指基團-ΝΗ2。「一級醇」係指基團-烷基-ΟΗ，其中羥基係連接至一級碳。實例包括-CH2〇H(羥甲基）、-CH2CH2OH(羥乙基）及 -CH(CH3)CH2〇H(l-羥丙-2_基）。「烯基」係指具有2至6個碳原子且較佳2至4個碳原子且具有至少1個且較佳1至2個乙烯基（>c=C<)不飽和位點之直鍵或分支鏈烴基。該等基團例示為乙烯基、烯丙基及丁 _3_ 稀-1-基。此術語中包括順式異構體及反式異構體或此等異構體之混合物。「快基」係指具有2至6個碳原子且較佳2至3個碳原子且具有至少1個且較佳！至2個乙炔系_CsC_不飽和位點之直鏈或分支鏈單價烴基。該等炔基之實例包含乙炔基（_CsCH) 及炔丙基（-CH2CsCH)。「羧基（Carboxy〖）」或「羧基（carboxy)」係指-COOH或其鹽。「氰基」或「腈」係指基團_CN。「環烷基」係、#具有單環之3至13個碳原子之環狀烷基。環烧基之實例包括環丙基、環丁基、環戊基、環辛基 159849.doc 201249786 及其類似基團。「胍基」係指基團-NHC(=NH)NH2。由基J或自素」係指乳、氣、漠及破，且通常為敗或氣。羥基（hydroxy)」或「羥基（hydr〇xyl)」係指基團_〇H。雜環」及「雜環基」係指具有單環且具有3至15個環原子，包括1至4個雜原子之飽和或不飽和基團。此等環原子係選自由氮、硫或氧組成之群。在一實施例中，雜環基團之氮及/或硫原子視情況氧化以提供N_氧化物、·s(〇)_ 或-S02-部分。「硝基」係指基團_N〇2 » 「亞硝基」係指基團_Ν0。「側氧基（oxo) J係指原子（=〇)。「經取代」係指一或多個氫經選自由以下組成之群的取代基置換之基團：烷氧基、醯基、醯胺基、醯氧基、胺基、胺基羰基、胺基硫羰基、胺基羰基胺基、胺基硫羰基胺基、胺基幾氧基、甲脒、基、缓基、叛基醋、（叛基醋）胺基、（羧基酯）氧基、氰基、胍基、齒基、羥基、硝基、 s〇3H、磺醯基、磺醯基氧基、硫醯基、硫醇及烷硫基，其中該等取代基係如本文所定義。在某些經取代之環狀基團中，「經取代」亦係指兩個氫經單個雙鍵氧原子（側氧基）或單個雙鍵硫原子（硫酮基）置換之基團。在一些實施例中，經取代之基團具有1至3個前述取代基。在其他實施例中’經取代之基團具有1至2個前述取代基。 159849.docC^C: 6 cycloalkyl, wherein at least one substituent is a Ci_C3 primary alcohol; B is absent, or is -CH=CH-, -C^C- or unsubstituted phenyl; c is CH CH- , -C=C- or unsubstituted phenyl, wherein if 6 is _ch=ch_, then c is not also _CH=CH_; R1, ... and 3 are independently selected from hydrogen and substituted or An unsubstituted Cl_C3 alkyl group, or Rl&R2 together with the carbon atom to which it is attached, forms an unsubstituted 6-cycloalkyl group, or r2 and r3 together with the carbon atom to which they are attached form 5 to 8 a substituted or unsubstituted heterocyclic ring of a ring atom wherein the (1) ring atom of the hetero ring is selected from the group consisting of N, fluorene and S; and (iv) (10), wherein the hydride is hydrogen or an unsubstituted c3 alkyl group. Another aspect of the diarrhea provides for the treatment of a patient having a Gram-negative bacterial infection by a co-administration of a synergistic amount, such as an in vivo synergistic amount of a bacterial agent and an LpxC inhibitor of Formula I. In one embodiment, the antimicrobial agent is freed from the group consisting of vancomycin, linoleamide, azithromycin, and amine. N. teicoplanin, daptomycin, clindamycin, rifa 159849.doc 201249786 Ping, cefotaxime, gentamicin, novomycin and telavancin. In one such embodiment, the antimicrobial agent is vancomycin or rifampicin. These and other aspects of the present invention will become apparent upon reference to the following embodiments. The present invention provides novel compounds, methods of inhibiting LpxC in Gram-negative bacteria, and novel methods of treating bacterial infections. The compounds provided herein can be formulated into pharmaceutical formulations and medicaments suitable for use in the methods of the invention. The invention also provides for the use of a compound in the manufacture of a medicament and a pharmaceutical formulation, the use of the compound in inhibiting LpxC, and the use of the compound in the treatment of bacterial infection in an individual. The present invention further provides a composition and method for treating Gram-negative infection by inhibiting UDp_3_〇_(R_3_hydroxyindenyl)-Ν-acetamid glucosamine deacetylase by separate administration ( The compound active as LpxC) is either achieved in combination with administration of the first antibacterial compound. A. Definitions The following abbreviations and definitions are used throughout this application: "LpxC" is an abbreviation for UDP_3_〇_(R_3_hydroxyindenyl)_N acetoglucoside deacetylase. Unless otherwise indicated, the following definitions shall apply as used herein. The "alkyl group" means a monovalent saturated aliphatic hydrocarbon group having 1 to 10 carbon atoms and preferably 1 to 6 carbon atoms. The term includes, for example, straight-chain and branched-chain hydrocarbon groups such as methyl (CH3.), ethyl (ch3ch2-), n-propyl (CH3CH2CH2-), isopropyl ((CH3)2CH-), n-butyl (CH3CH2CH2CH2_). ), isobutyl ((CHACHCH2·), second butyl ((CH3)(CH3CH2)CH), third 159849.doc 201249786 ((CH3)3C-), n-pentyl (ch3ch2ch2ch2ch2-) and new Pentyl ((CH3)3CCH2-). "Alkoxy" refers to the group - oxime-alkyl, wherein the alkyl group is as defined herein. The alkoxy group includes a decyloxy group, an ethoxy group, a n-propoxy group. , isopropoxy, n-butoxy, tert-butoxy, second butoxy, n-pentyloxy and the like. "Amine" refers to the group -ΝΗ2. "Primary alcohol" means a group -alkyl-hydrazine wherein the hydroxy group is attached to the primary carbon. Examples include -CH2〇H (hydroxymethyl), -CH2CH2OH (hydroxyethyl), and -CH(CH3)CH2〇H (l-hydroxypropyl- "Alkenyl" means "having 2 to 6 carbon atoms and preferably 2 to 4 carbon atoms and having at least 1 and preferably 1 to 2 vinyl groups (>c=C<) a direct or branched hydrocarbon group at a saturated site. These groups are exemplified as vinyl or ally. And _3_ dimethyl-1-yl. This term includes cis isomer and trans isomer or a mixture of such isomers. "Fast radical" means having 2 to 6 carbon atoms and preferably a linear or branched chain monovalent hydrocarbon group of 2 to 3 carbon atoms and having at least one and preferably! to 2 acetylene-CsC_unsaturated sites. Examples of such alkynyl groups include ethynyl (_CsCH) and alkyne Propyl (-CH2CsCH) "Carboxy" or "carboxy" means -COOH or a salt thereof. "Cyano" or "nitrile" means a group -CN. "Cycloalkyl" And a cyclic alkyl group having 3 to 13 carbon atoms in a single ring. Examples of the cycloalkyl group include a cyclopropyl group, a cyclobutyl group, a cyclopentyl group, a cyclooctyl group 159849.doc 201249786 and the like. "Alkyl" refers to the group -NHC(=NH)NH2. The term "J or J" refers to milk, gas, and moisture, and is usually ruin or gas. Hydroxyl or hydroxy "xyl)" refers to the group _〇H. Heterocycle" and "heterocyclyl" means a saturated or unsaturated group having a single ring and having 3 to 15 ring atoms, including 1 to 4 hetero atoms. Equivalent atomic system a group of free nitrogen, sulfur or oxygen. In one embodiment, the nitrogen and/or sulfur atom of the heterocyclic group is optionally oxidized to provide an N-oxide, ·s(〇)_ or -S02- moiety. "Nitro" means a group _N〇2 » "Nitroso" means a group _Ν0. "Oxyl group (oxo) J means an atom (=〇). "Substituted" means a group in which one or more hydrogens are replaced by a substituent selected from the group consisting of an alkoxy group, a decyl group, a decylamino group, a decyloxy group, an amine group, an amine carbonyl group, an amine group. Thiocarbonyl, aminocarbonylamino, aminothiocarbonylamino, aminooxy, formazan, sulfhydryl, thioglycolic acid, thioglycolic acid, (carboxy ester)oxy, cyanide Base, sulfhydryl, dentate, hydroxy, nitro, s〇3H, sulfonyl, sulfonyloxy, thiol, thiol and alkylthio, wherein the substituents are as defined herein. In some substituted cyclic groups, "substituted" also refers to a group in which two hydrogens are replaced by a single double bond oxygen atom (sideoxy group) or a single double bond sulfur atom (thiol group). In some embodiments, the substituted group has from 1 to 3 of the foregoing substituents. In other embodiments, the substituted group has from 1 to 2 of the foregoing substituents. 159849.doc

S 12 201249786 墙酿基」係指基團_s〇2_烧基、經取代之烧基、 -so2-烯基、_犯2_經取代之稀基，其中烧基、經取代之烧基稀基 '經取代之婦I、炔基及經取代之炔基係如本文所定義。磺醯基包括諸如甲基-so2-之基團。確酿基氧基」係指基團_〇s〇2烧基、〇s〇2經取代之烷基os〇2_烯基、-oso2-經取代之烯基、_os〇2_炔基〇s〇2_經取代之炔基，其中烷基、經取代之烷基、烯基、經取代之烯基、炔基及經取代之炔基係如本文所定義。「硫醯基」係指基團H-C(s)-、烷基-C(s)-、經取代之烷基-C(s)-、烯基_c(S)-、經取代之烯基_c(s)_、炔基-C(S)- 及經取代之炔基-C(s)_，其中烷基、經取代之烷基、烯基、經取代之烯基、炔基及經取代之炔基係如本文所定義。「硫醇」係指基團-SH。「硫酮基」係指原子（=s)» 「燒硫基」係指基團-s_烷基，其中烷基係如本文所定義°在其他實施例中，硫可氧化成_s(0)_。亞砜可以一或多種立體異構體形式存在。除非另外指示，否則本文未明確定義之取代基的命名係藉由命名官能基之末端部分，之後朝向連接點命名相鄰官能基來達成。舉例而言，取代基「芳基烷氧基羰基」係指基團（芳基）-(烷基）-o-c(o)-。通常’提及諸如氫或Η之某一元素意謂包括該元素之所 159849.doc •13- 201249786 有同位素。舉例而若取代基㈣義包括氫如，則其亦包括氘及氚。、本發明亦包括同位素標記之本發明化合物，該等同位素 ‘ s己之本發明化合物與本文所揭示之化合物在結構上相同但事實上，一或多個原子經具有與通常見於自缺界之原子質量或質量數不同之原子質量或質量數的原子置換。可併入本發明化合物中之同位素之實例包括氫、碳、氮、氧碌、硫、乳及氣之同位素，分別諸如2h、4、％、 C、15N、18〇、Po、3 丨p、、 14 32 35S 12 201249786 "Wheat base" refers to the group _s〇2_alkyl, substituted alkyl, -so2-alkenyl, _ 2 - substituted dilute, wherein alkyl, substituted alkyl The dilute base 'substituted women I, alkynyl and substituted alkynyl are as defined herein. The sulfonyl group includes a group such as methyl-so2-. "Acceptable oxy" refers to the group _〇s〇2 alkyl, 〇s〇2 substituted alkyl os〇2_alkenyl, -oso2-substituted alkenyl, _os〇2_alkynyl fluorene S〇2_substituted alkynyl wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl and substituted alkynyl are as defined herein. "Thioinyl" refers to the group HC(s)-, alkyl-C(s)-, substituted alkyl-C(s)-, alkenyl-c(S)-, substituted alkenyl _c(s)_, alkynyl-C(S)- and substituted alkynyl-C(s)_, wherein alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl and Substituted alkynyl groups are as defined herein. "Mercaptan" refers to the group -SH. "Thionyl" means an atom (=s)» "Sulfuryl" refers to the group -s-alkyl, wherein alkyl is as defined herein. In other embodiments, sulfur can be oxidized to _s ( 0)_. The sulfoxide may exist in one or more stereoisomers. Unless otherwise indicated, the nomenclature of a substituent not specifically defined herein is achieved by naming the terminal portion of the functional group and then naming the adjacent functional group toward the point of attachment. For example, the substituent "arylalkoxycarbonyl" refers to the group (aryl)-(alkyl)-o-c(o)-. Usually, the reference to an element such as hydrogen or helium means that the element is included. 159849.doc •13- 201249786 There are isotopes. For example, if the substituent (IV) includes hydrogen, then it also includes ruthenium and osmium. The invention also includes isotopically-labeled compounds of the invention which are structurally identical to the compounds disclosed herein but in fact one or more of the atoms are typically found in the self-deficient Atomic substitution of atomic mass or mass number with different atomic mass or mass number. Examples of isotopes which may be incorporated into the compounds of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen, sulfur, milk and gas, such as 2h, 4, %, C, 15N, 18, Po, 3 丨p, , 14 32 35

P s、18f&36c卜含有前述二素及/或其他原子之其他同位素之本發明化合物、其前藥及該等化合物及該等前藥之醫藥學上可接受之鹽在本發明之㈣内。本發明之某些同位素標記之化合物，例如併有諸如加"C之放射性同位素之化合物適用於藥物及/ 或基質組織分佈分析。氣化同位素（亦即3H)及碳14同位素 (亦即14C)因其易於製備及可偵測而尤其較佳。此外經諸如氛（亦即2H)之較重同位素取代可由於較大之代謝穩定性而提供某些治療優勢，例如增加之活體内半衰期或降低之劑量需求’因此可在某些情況下較佳。本發明之同位素標記之化合物及其前藥-般可藉由進行已知或引用的程序: 藉由以易獲得之同位素標記之試劑取代未經同位素標記之試劑來製備。「立體異構體（stereoisomer)」係指具有相同原子連接性但具有不同原子空間排列的化合物。立體異構體包括順式·反式異構體、E&Z異構體、對映異構體及非對映異構 159849.doc 14· 201249786 體。互變異構艘」係指質子位置不同之分子替換形式，諸如烯醇-酮基及亞胺-烯胺互變異構體，或含 NH-環原子排列之雜芳基之互變異構形式，諸如吡唑、咪唑、苯并咪唑、***及四唑。一般技術者認為可能存在其他互變異構環原子排列。「患者」係指人類及非人類動物，尤其是哺乳動物。醫藥學上可接受之里」係指化合物之醫藥學上可接受之鹽，該等鹽衍生自此項技術中熟知之多種有機及無機相對離子且包括（僅舉例而言）納、钟、約、鎮、録、四燒基銨及類似物；及（當分子含有鹼式官能基時）有機酸或無機酸之鹽，諸如鹽酸鹽、氫溴酸鹽、灑石酸鹽、甲磺酸鹽、乙酸鹽、順丁稀一酸鹽、乙二酸鹽、礙酸鹽、硫酸鹽及其類似鹽。醫藥學有效量」及「治療有效量」係指足以治療特定病症或疾病或一或多種其症狀及/或預防該疾病或病症發生之化合物之量。如本文所用，術語「協同作用」或「協同」意謂化合物在組合使用時之組合作用大於化合物個別使用時之相加作用。「協同作用」可定量定義為部分抑制濃度指數 (FICDM.5，其中FICI定義為兩種化合物之組合中個別組分之部分抑制濃度（FIC)之總和，且FIC定義為組合中化合物之最低抑制濃度（MIC)除以單獨化合物之MIC之比率： 159849.doc -15· 201249786 pjQj — (MiC組合中之藥物A λ + (MJC組合中之藥物b \ \ MJC!單獨藥物A / V财單獨藥物B / 或者’「協同作用」’更特定言之「活體内協同作用」可疋量疋義為與單獨LpxC抑制劑或第二抗菌劑相比，組八使用之藥劑之靜態劑量（Static dose)減少到至多1/2。在某此情況下，單獨一種藥劑可能決達不到靜態劑量。在該等情況下，若細菌生長可藉由組合投與兩種單獨不能達成停滯之化合物而停止（與感染後〇小時所量測一致之24小時時之 CFU負荷），則組合為協同組合。「共同投與」可呈單一調配物形式（組合例如本發明化合物與第二抗菌劑以及醫藥學上可接受之賦形劑，視情況於經設計為獨立控制兩種活性成分之各別釋放速率及^續時間之不同賦形劑混合物中分隔兩種活性成分）或藉由獨立投與含有活性劑之各別調配物來實現。「共同投與」進一步包括並行投與（同時投與本發明化合物及第二抗菌劑）及隨時間變化投與(在與投與第二抗菌劑不同之時間投與本發明化合物），只要本發明化合物與第二抗菌劑在至少部分重叠之時間期間以治療上有效濃度均存在於身體可〇「術抗菌劑」係指具有殺菌或抑菌活性之藥劑。術語「抑制生長」表示特定細菌群體之數目的增長速率降低。因此，該術語包括細菌群體增加但增加速率降低之情形，以及群體生長停止之情形，以及群體中細菌數減少或甚至群體消除之情形。若使用酶活性分析來筛選抑制劑，則可 159849.docP s, 18f & 36c, a compound of the invention, a prodrug thereof, and the pharmaceutically acceptable salts of the compounds and the prodrugs of the invention, wherein the pharmaceutically acceptable salt of the compound and the other isotopes of the other atom are within the invention . Certain isotopically-labeled compounds of the invention, e.g., compounds having a radioisotope such as "C, are suitable for drug and/or matrix tissue distribution analysis. The gasified isotope (i.e., 3H) and carbon 14 isotope (i.e., 14C) are particularly preferred because of their ease of preparation and detection. In addition, heavier isotopic substitutions such as ambience (i.e., 2H) may provide certain therapeutic advantages due to greater metabolic stability, such as increased in vivo half-life or reduced dosage requirements, and thus may be preferred in some cases. . The isotope-labeled compounds of the present invention and prodrugs thereof can be prepared by a known or cited procedure: by substituting an easily isotope-labeled reagent for an unisotopically labeled reagent. "stereoisomer" means a compound having the same atomic connectivity but having a different arrangement of atoms. Stereoisomers include cis-trans isomers, E&Z isomers, enantiomers, and diastereoisomers 159849.doc 14·201249786. "Tautomeric ship" means a molecular replacement form having a different proton position, such as an enol-keto group and an imine-enamine tautomer, or a tautomeric form of a heteroaryl group containing an NH-ring atom arrangement, such as Pyrazole, imidazole, benzimidazole, triazole and tetrazole. The average skilled person believes that there may be other tautomeric ring atom arrangements. "Patient" means human and non-human animals, especially mammals. "Pharmaceutically acceptable" means a pharmaceutically acceptable salt of a compound derived from a variety of organic and inorganic counterions well known in the art and including, by way of example only, , town, recorded, tetraalkylammonium and the like; and (when the molecule contains a basic functional group) a salt of an organic or inorganic acid, such as a hydrochloride, a hydrobromide, a tartrate, methanesulfonic acid Salt, acetate, cis-butylate, oxalate, acid salt, sulfate and the like. "Pharmaceutically effective amount" and "therapeutically effective amount" mean an amount of a compound which is sufficient to treat a particular condition or disease or one or more of its symptoms and/or prevent the occurrence of the disease or condition. As used herein, the term "synergistic" or "synergistic" means that the combined action of the compounds when used in combination is greater than the additive effect of the individual use of the compounds. "Synergy" can be quantitatively defined as a partial inhibitory concentration index (FICDM.5, where FICI is defined as the sum of the partial inhibitory concentrations (FIC) of the individual components of the combination of the two compounds, and FIC is defined as the minimum inhibition of the compound in the combination. Concentration (MIC) divided by the ratio of the MIC of the individual compound: 159849.doc -15· 201249786 pjQj — (The drug A λ + in the MiC combination (drug in the MJC combination b \ \ MJC! Drug A / V alone drug alone) B / or '"Synergy"' more specifically "in vivo synergy" can be measured as a static dose of the agent used in Group 8 compared to a single LpxC inhibitor or a second antibacterial agent. Reduce to a maximum of 1/2. In some cases, a single agent may never reach a static dose. In such cases, if the growth of the bacteria can be stopped by combining two compounds that cannot achieve stagnation alone ( The CFU load at 24 hours, which is consistent with the measurement after the infection, is combined into a synergistic combination. The "co-administered" may be in the form of a single formulation (combination of, for example, the compound of the present invention and the second antibacterial agent) And pharmaceutically acceptable excipients, as appropriate, separate the two active ingredients in separate excipient mixtures designed to independently control the respective release rates and durations of the two active ingredients) or by independent injection This is achieved with separate formulations containing the active agent. "Co-administered" further includes concurrent administration (simultaneous administration of the compound of the invention and the second antimicrobial agent) and administration over time (in the administration of a second antimicrobial agent) The compound of the present invention is administered at different times, as long as the compound of the present invention and the second antibacterial agent are present in the body at a therapeutically effective concentration for at least a portion of the time period, and the bactericidal or bacteriostatic activity is present in the body. The term "inhibiting growth" means a decrease in the rate of growth of the number of specific bacterial populations. Thus, the term includes the case where the bacterial population increases but the rate of increase decreases, as well as the situation in which the population growth ceases, and the number of bacteria in the population is reduced or even Eliminate the situation. If using enzyme activity analysis to screen for inhibitors, then 159849.doc

S -16 - 201249786 對化合物之攝取/流出、溶解度、半衰期等作出修改以使酶抑制與生長抑制關聯。抗菌劑活性不一定限於細菌，作亦可涵蓋針對寄生蟲、病毒及真菌之活性。除非上下文另外需要，否則在整個說明書及隨後之申請專利範圍中，措詞「包含（comprise)J及其變體（諸如「包含（comprises)」及「包含（c〇mpriseing)」）將以開放包含性意義來解釋，亦即解釋為「包括（但不限於）」。在整個本說明書令提及「一實施例（〇ne emb〇diment)」或「一實施例（an embodiment)」意謂結合該實施例所描述之特定特徵、結構或特性包括於本發明之至少一實施例中。因此，在整個本說明書中各處出現片語「在一實施例中J不-定均指相同實施例。另夕卜可在一或多個實施例中以任何適合方式組合特定特徵、結構或特性。 B.化合物、組合物及其用途在一態樣中，本發明提供式！化合物：S -16 - 201249786 Modifications to the uptake/efflux, solubility, half-life, etc. of the compounds to correlate enzyme inhibition with growth inhibition. The activity of the antibacterial agent is not necessarily limited to bacteria, and may also cover activities against parasites, viruses, and fungi. Unless the context requires otherwise, the wording "comprise" J and its variants (such as "comprises" and "includes (c〇mpriseing)") will be open throughout the specification and subsequent claims. Interpretative meaning is interpreted as “including (but not limited to)”. References throughout the specification to "an embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in the invention. In an embodiment. Thus, the phrase "a" or "an embodiment" is used throughout the specification to refer to the same embodiment. In one or more embodiments, a particular feature, structure or combination may be combined in any suitable manner. B. Compounds, Compositions, and Uses Thereof In one aspect, the invention provides a compound of formula:

及其立體異構體及醫藥學上可接受之鹽，其中A為經取代之c3_C6m其中至少—個取代基為ca —級醇；b 不存在，或為-CH=CH-、_c心或未經取代之苯基；c為 H C-C-或未經取代之苯基，其中若β為則C不會亦為·chsch-; R1、R2及R3係獨立地選自氫及 159849.doc •17· 201249786 經取代或未經取代之Cl_C3烷基，或Ri及R2連同其所連接之碳原子一起形成未經取代之C3_C6環烷基，或R2及尺3連同其所連接之碳原子及Q 一起形成具有5至8個環原子之經取代或未經取代之雜環，其中該雜環之i至2個環原子係選自N、0及s ;且Q為〇4NR，其中R為氫或未經取代之Ci_ c3烷基。And stereoisomers thereof and pharmaceutically acceptable salts thereof, wherein A is substituted c3_C6m wherein at least one of the substituents is a ca-alcohol; b is absent, or is -CH=CH-, _c heart or not Substituted phenyl; c is H CC- or unsubstituted phenyl, wherein if C is, then C will not be also chsch-; R1, R2 and R3 are independently selected from hydrogen and 159849.doc • 17 · 201249786 Substituted or unsubstituted Cl_C3 alkyl, or Ri and R2 together with the carbon atom to which they are attached form an unsubstituted C3_C6 cycloalkyl group, or R2 and 尺3 together with the carbon atom to which they are attached and Q Forming a substituted or unsubstituted heterocyclic ring having 5 to 8 ring atoms, wherein i to 2 ring atoms of the heterocyclic ring are selected from N, 0 and s; and Q is 〇4NR, wherein R is hydrogen or Unsubstituted Ci_ c3 alkyl.

在某些實施例中，Q為NR，且在一些該等實施例中，Q 為NH或NCH3。在某些實施例中，Ri、“及尺3係獨立地選自氫及經取代或未經取代之C〗_C3烷基，且在一些該等實施例中係選自氫及未經取代之Ci_C3烷基。在某些實施例中，y及R2獨立地為未經取代之Cl·。烷基。在某些實施例中，A為經羥甲基取代之C3_C6環烷基。在其他實施例中，B及C為-CsC-。本發明之一態樣提供式〗-八化合物：In certain embodiments, Q is NR, and in some such embodiments, Q is NH or NCH3. In certain embodiments, Ri, "and Ruler 3 are independently selected from hydrogen and substituted or unsubstituted C"-C3 alkyl, and in some such embodiments are selected from hydrogen and unsubstituted. Ci_C3 alkyl. In certain embodiments, y and R2 are independently unsubstituted C. alkyl. In certain embodiments, A is hydroxymethyl substituted C3_C6 cycloalkyl. In the examples, B and C are -CsC-. One aspect of the present invention provides a formula - eight compounds:

及其立體異構體及醫藥學上可接受之鹽，其中八為經取代之Cs-C6環烷基，其中至少一個取代基為Ci_C3__級醇； R、R及R係獨立地選自氫及經取代或未經取代之C广q 烷基，或R1及R2連同其所連接之碳原子一起形成匚广匕環烷基，或R2及R3連同其所連接之碳原子及氮一起形成具有 5至8個環原子之經取代或未經取代之雜環，其中雜環之！ J59849.doc _18_And stereoisomers thereof and pharmaceutically acceptable salts thereof, wherein eight are substituted Cs-C6 cycloalkyl groups, at least one of which is a Ci_C3__ alcohol; R, R and R are independently selected from hydrogen And a substituted or unsubstituted C-g-Q-alkyl group, or R1 and R2 together with the carbon atom to which they are attached form a fluorene-cycloalkyl group, or R2 and R3 together with the carbon atom to which they are attached and nitrogen form a substituted or unsubstituted heterocyclic ring of 5 to 8 ring atoms, of which a heterocyclic ring! J59849.doc _18_

S 201249786 至2個環原子係選自N、〇及S ;且R為氫或未經取代之Ci_ A烧基。在某些實施例中，Ri、以及尺3係獨立地選自氣及經取代或未經取代之Cl_C3烷基，且在—些該等實施例中S 201249786 to 2 ring atoms are selected from N, fluorene and S; and R is hydrogen or an unsubstituted Ci_ A alkyl group. In certain embodiments, Ri, and Caliper 3 are independently selected from the group consisting of a gas and a substituted or unsubstituted C1-C3 alkyl group, and in some of these embodiments

159849.doc .19· 201249786159849.doc .19· 201249786

表I ,〇2 R3 o R1\/ / I 〇 A B C R1 R2 RJ Q 1-7 HO-^ 〇-i ch3 ch3 CH2CH3 NH 1-8 HO-^ ch3 ch3 CH2CH20H NH 1-9 HO-\ ch3 ch3 ch3 nch3 1-10 HO-^ ch3 ch3 H 0 1-11 Λ HO ch3 ch3 H NH 1-12 丨 HO ch3 ch3 H NH 1-13 HO ch3 ch3 H NH 1-14 HO^X- * ch3 ch3 H NH 1-15 HO N-f ch3 ch3 H NH 1-16 /丨丨丨__丨"1 HO 、-’ ch3 ch3 H NH 1-17 HO-\ 卜1 環丁基 H NH 159849.doc -20- s 201249786Table I, 〇2 R3 o R1\/ / I 〇ABC R1 R2 RJ Q 1-7 HO-^ 〇-i ch3 ch3 CH2CH3 NH 1-8 HO-^ ch3 ch3 CH2CH20H NH 1-9 HO-\ ch3 ch3 ch3 Nch3 1-10 HO-^ ch3 ch3 H 0 1-11 Λ HO ch3 ch3 H NH 1-12 丨HO ch3 ch3 H NH 1-13 HO ch3 ch3 H NH 1-14 HO^X- * ch3 ch3 H NH 1 -15 HO Nf ch3 ch3 H NH 1-16 /丨丨丨__丨"1 HO ,-' ch3 ch3 H NH 1-17 HO-\ Bu 1 Cyclobutyl H NH 159849.doc -20- s 201249786

本發明化合物可輕易地使用本文所述之方法或此項技術中熟知之其他方法合成。舉例而言，異羥肟酸或具有多種取代基之類似架構之合成廣泛查閱Kline，T.等人，「Potent, novel in vitro inhibitors of the Pseudomonas aeruginosa deacetylase LpxC」《/. Mec? C/zem. 2002, 45(14), 3112-29 ；美國專利第5,925,659號；Pimmg，M. C.等人，「A Convenient Procedure for the Preparation of Amino Acid Hydroxamate from Esters」·/· Chem. 1995, 60, 8084- 159849.doc •21- 201249786 8085 ; Nhu，K.等人，「A New and Efficient Solid Phase Synthesis of Hydroxamic Acids」J_ Chem. 1997，62, 7088-7089 ；國際PCT公開案第 W098/18754號；Mellor，S. L·，等人，「N-Fmoc-aminoxy-2-chlortrityl Polystyrene Resin: A Facile Solid-phase Methodology for the Synthesis of Hydroxamic Acids」 Tetrahedron Lett. 1997, 3 8, 3311-3314 i Khan, S. I·等人，「A Facile and Convenient Solid- phase Procedure for Synthesizing Nucleoside Hydroxamic Acids」 Tetrahedron. Lett. 1998，39，8031-8034 ; Zhang, Y. 等人，「Design，Combinatorial Chemical Synthesis, and in vitro Characterization of Novel Urea Based Gelatinase Inhibitors」Med C/ze/w. Ze". 1999，9，2823-2826 ; Ito，Y.等人，「Synthetic Reactions by Complex Catalysts. XXXI， A Novel and Versatile Method of Heterocycle Synthesis」·/. C/zewi· iSoc. 1973，95，4447-4448 ; Ito，Y. 等人，「Synthetic Reactions by Complex Catalysts XXXV」 Gom/nww. 1974, 4, 97-103 ; Witte，H.等人，rGyclische Imidsaurester aus Nitrilen und Aminoalkoholen」 C/ze/w. 1974，996-1009 ; Pattenden, G.等人，「Naturally Occurring Linear Fused Thiazoline-Thiazole Containing Metabolites: Total Synthesis of (-)The compounds of the invention can be readily synthesized using the methods described herein or other methods well known in the art. For example, the synthesis of hydroxamic acids or similar structures with multiple substituents is extensively reviewed by Kline, T. et al., "Potent, novel in vitro inhibitors of the Pseudomonas aeruginosa deacetylase LpxC" /. Mec? C/zem. 2002, 45(14), 3112-29; U.S. Patent No. 5,925,659; Pimmg, MC et al., "A Convenient Procedure for the Preparation of Amino Acid Hydroxamate from Esters"·/· Chem. 1995, 60, 8084- 159849. Doc •21-201249786 8085; Nhu, K. et al., “A New and Efficient Solid Phase Synthesis of Hydroxamic Acids” J_Chem. 1997, 62, 7088-7089; International PCT Publication No. W098/18754; Mellor, S L., et al., "N-Fmoc-aminoxy-2-chlortrityl Polystyrene Resin: A Facile Solid-phase Methodology for the Synthesis of Hydroxamic Acids" Tetrahedron Lett. 1997, 3 8, 3311-3314 i Khan, S. I · et al., "A Facile and Convenient Solid-phase Procedure for Synthesizing Nucleoside Hydroxamic Acids" Tetrahedron. Lett. 1998, 39, 8031-8034; Zhang, Y. et al., "Design, Combinatorial Chemica l Synthesis, and in vitro Characterization of Novel Urea Based Gelatinase Inhibitors" Med C/ze/w. Ze". 1999, 9, 2823-2826; Ito, Y. et al., "Synthetic Reactions by Complex Catalysts. XXXI, A Novel And Versatile Method of Heterocycle Synthesis"·/. C/zewi· iSoc. 1973, 95, 4447-4448; Ito, Y. et al., "Synthetic Reactions by Complex Catalysts XXXV" Gom/nww. 1974, 4, 97-103 Witte, H. et al., rGyclische Imidsaurester aus Nitrilen und Aminoalkoholen" C/ze/w. 1974, 996-1009; Pattenden, G. et al., "Naturally Occurring Linear Fused Thiazoline-Thiazole Containing Metabolites: Total Synthesis of (- )

Didehydromirabazole A, a Cytotoxic Alkaloid from Blue- 1636 ; Boyce，R. J.等人，「Total Synthesis of Thiangazole， 159849.doc -22- s 201249786 A Novel Naturally Occurring HIV-1 Inhibitor from Polyangium sp.」Tetrahedron 1995， 51， 7321-7330 ;Didehydromirabazole A, a Cytotoxic Alkaloid from Blue- 1636; Boyce, RJ, et al., "Total Synthesis of Thiangazole, 159849.doc -22- s 201249786 A Novel Naturally Occurring HIV-1 Inhibitor from Polyangium sp." Tetrahedron 1995, 51, 7321 -7330;

Galeotti，N.等人，「Synthesis of Peptidyl Aldehydes from Thiazolines」Tetrahedron. Lett. 1997, 38, 2459-2462 ; Charette，A. B.等人，「Mild Method for the Synthesis of Thiazolines from Secondary and Tertiary Amides」J. Org. Chem. 1998，63, 908-909 ； Bergeron, R. J.等人，「Effects of C-4 Stereochemistry and C-4' Hydroxylation on the Iron Clearing Efficiency and Toxicity of Desferrithiocin Analogues」·/· Med. C/ie/n. 1999，42，2432-2440 ; Raman, P. 等人，「Titanium (IV)-mediated Tandem Deprotection-cyclodehydration of Protected Cysteine N-Amides: Biomimetic Synthesis of Thiazoline- and Thiazole-containing Heterocycles」 Og. Zeii. 2000, 2, 3289-3292 ; Fernandez, X.等人，「Novel Synthesis of 2-Thioazolines」Tetrahedron Lett, 2000，41， 3381-3384 ;及 Wipf， P.等人，「C. Thiolysis ofGaleotti, N., et al., "Synthesis of Peptidyl Aldehydes from Thiazolines" Tetrahedron. Lett. 1997, 38, 2459-2462; Charette, AB et al., "Mild Method for the Synthesis of Thiazolines from Secondary and Tertiary Amides" J. Org Chem. 1998, 63, 908-909; Bergeron, RJ, et al., "Effects of C-4 Stereochemistry and C-4' Hydroxylation on the Iron Clearing Efficiency and Toxicity of Desferrithiocin Analogues"·/· Med. C/ie/ n. 1999, 42, 2432-2440; Raman, P. et al., "Titanium (IV)-mediated Tandem Deprotection-cyclodehydration of Protected Cysteine N-Amides: Biomimetic Synthesis of Thiazoline- and Thiazole-containing Heterocycles" Og. Zeii. 2000, 2, 3289-3292; Fernandez, X. et al., "Novel Synthesis of 2-Thioazolines" Tetrahedron Lett, 2000, 41, 3381-3384; and Wipf, P. et al., "C. Thiolysis of

Oxazolinenes: A New, Selective Method for the Direct Conversion of Peptide Oxazolines into Thiazolines」 Z/g". 1995，36，6395-6398，該等文獻以引用的方式併入本文中β 在另一態樣中，本發明提供一種包含式I化合物或其立體異構體或醫藥學上可接受之鹽及醫藥學上可接受之載劑或稀釋劑的醫藥組合物。在另一態樣中，本發明提供一種抑制革蘭氏陰性細菌中 159849.doc -23· 201249786 去乙醯基酶’藉此影響細菌生長之方法，其包含向需要該抑制之患者投與式I化合物或其立體異構體或醫藥學上可接受之鹽。在另一態樣中，本發明提供一種抑制革蘭氏陰性細菌中 LpxC ,藉此調整細菌感染之毒性之方法，其包含向需要該抑制之患者投與式I化合物或其立體異構體或醫藥學上可接觉之鹽。在使用本發明化合物抑制LpxC2方法之某些實施例中，相對於LpxC，化合物之1〇^值小於或等於1〇 μΜ。在其他實施例中，1(：5〇值小於或等於i μΜ，小於或等於〇_1 μΜ，小於或等於〇〇5〇 μΜ，小於或等於〇〇3〇，小於或等於0.025 μΜ，或小於或等於〇〇1〇 μΜ。在另一態樣中，本發明提供一種治療患有革蘭氏陰性細菌感染之個體的方法，其包含向有需要個體投與抗菌有效量之式I化合物或其立體異構體或醫藥學上可接受之鹽。在一個該實施例中’細菌為綠膿假單胞菌、伯克霍爾德氏菌屬(例如孕鹿㈣賞摩德磨)、腸桿菌科、弗朗西絲菌屬 (例如上拉癬弟苈西.絲磨）、沙雷氏菌屬、變形桿菌屬、克雷伯氏菌！、腸桿菌屬、檸檬酸桿菌屬、沙門氏菌、普羅威登斯菌屬、耶氏桿菌屬（例如Μ顯#磨）、摩根氏菌屬或大腸桿菌。在一特定實施例中’細磨為綠膿假單胞菌、伯克霍爾德氏菌屬、弗朗西絲菌屬、腸桿菌屬、耶氏柃菌屬或大腸桿菌。在一個該實施例中，細菌為綠膿假單胞菌。在另-該實施例中、細菌為大腸桿菌。在另一實施賊今麥芽窄食單胞菌、木糖氡化產鹼菌、嗜血 159849.doc</ RTI> <RTIgt; The invention provides a pharmaceutical composition comprising a compound of formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. In another aspect, the invention provides a method of inhibiting 159849.doc -23· 201249786 deacetylase in Gram-negative bacteria, thereby affecting bacterial growth, comprising administering to a patient in need of such inhibition A compound or a stereoisomer thereof or a pharmaceutically acceptable salt thereof. In another aspect, the invention provides a method of inhibiting LpxC in a Gram-negative bacterium, thereby modulating the toxicity of a bacterial infection, comprising administering to a patient in need of such inhibition a compound of Formula I or a stereoisomer thereof or Pharmaceutically sensible salt. In certain embodiments of the method of inhibiting LpxC2 using a compound of the invention, the compound has a value of less than or equal to 1 〇 μΜ relative to LpxC. In other embodiments, 1 (:5〇 value is less than or equal to i μΜ, less than or equal to 〇_1 μΜ, less than or equal to 〇〇5〇μΜ, less than or equal to 〇〇3〇, less than or equal to 0.025 μΜ, or In another aspect, the invention provides a method of treating an individual having a Gram-negative bacterial infection comprising administering to the individual in need thereof an antibacterial effective amount of a compound of formula I or a stereoisomer or a pharmaceutically acceptable salt thereof. In one such embodiment, the bacterium is Pseudomonas aeruginosa, Burkholderia (eg, pregnant deer (four) musing mill), intestine Bacillus, Francis (eg, Lai, Daisy, Silk), Serratia, Proteus, Klebsiella, Enterobacter, Citrobacter, Salmonella, Providen Phytophthora, Yersinia (eg, sputum), Morganella or Escherichia coli. In a particular embodiment, 'fine grinding is Pseudomonas aeruginosa, Burkholderia, Francis, Enterobacter, Yarrowia or Escherichia coli. In one In this embodiment, the bacterium is Pseudomonas aeruginosa. In another embodiment, the bacterium is Escherichia coli. In another embodiment, the genus Stenotrophomonas maltophilia, the xylose sputum, the bacterium Blood 159849.doc

S •24- 201249786 桿菌屬、奈瑟菌屬、西地西菌屬I愛德華氏菌屬。在某些實施例中’個體可為哺乳動物，且在一些實施例中為人類。對本發明之治療敏感之細菌感染包括由以下引起之初次感染（primary infection)及共感染（co_infecti〇n):細菌物種及或多種其他感染物（infectious agent)，諸如細菌、病毒、寄生蟲及真菌。本發明化合物可用於治療由内毒素之細菌產生，且特別是由革蘭氏陰性細菌及在脂多醣（Lps)或内毒素之生物合成中使用LpxC之細菌所引起之病狀。本發明化合物亦適用於治療由脂質入及Lps或内毒素之細菌產生所引起或惡化之病狀，諸如敗血症、敗血性休克、全身發炎、局部發炎、慢性阻塞性肺病（c〇pD)及慢性支氣管炎之急性惡化（AECB)。對於此等病狀，治療包括投與本發明化合物或本發明化合物之組合，視情況連同第二藥劑，其中該第二藥劑為第二抗菌劑或非抗菌劑。對於敗血症、敗血性休克、全身發炎、局部發炎、慢性阻塞性肺病（COPD)及慢性支氣管炎之急性惡化（AECB)，代表性非抗菌劑包括抗内毒素，包括内毒素受體結合抗體、内毒素結合抗體、抗CD14結合蛋白抗體、抗脂多醣結合蛋白抗體及酪胺酸激酶抑制劑。在嚴重或慢性呼吸道感染之治療中，本發明化合物亦可與經由吸入投與之非抗菌劑一起使用。在此治療中所用之代表性非抗菌劑包括消炎性類固醇、非類固醇消炎劑、支 159849.doc •25- 201249786 氣管擴張藥、黏液溶解藥、抗哮喘治療劑及肺液界面活性劑。詳言之，非抗菌劑可為舒喘寧（albuterol)、沙丁胺醇 (salbuterol)、布***（budesonide)、倍氣米松 (beclomethasone)、***（dexamethasone)、奈多羅米 (nedocromil)、倍氣米松、I替卡松（fluticasone)、氟尼縮松（flunisolide)、曲安西龍（triamcinolone)、布洛芬 (ibuprofin)、羅非昔布（rofecoxib)、萘普生（naproxen)、塞内昔布（celecoxib)、奈多羅米（nedocromil)、異丙托敍 (ipratropium)、間經異丙腎上腺素（metaproterenol)、11 比布特羅（pirbuterol)、沙美特羅（salmeterol)、福莫特羅 (formoterol)、茚達特羅（indacaterol)、支氣管擴張藥、黏液溶解藥、卡夫康坦（calfactant)、貝拉康坦（beractant)、阿法泊拉康坦（poractant alfa)、舒伐辛（surfaxin)或百慕時 (pulmozyme)(亦稱為阿法鍵道酶（domase alfa))。本發明化合物可單獨或與第二抗菌劑組合用以治療嚴重或慢性呼吸道感染，包括嚴重肺及醫院感染，諸如由以下所引起之感染：羞藏I#磨（五《化rMacier flerogewe·?)、# 溝腸桿菌{Enterobacter cloacae)、九豫释魯、肺炎克雷伯氏菌（Klebsiella pneumoniae、、產酸克雷伯氏菌[Klebsiella oxytoca)、奇異變形捍菌（Proteus mirabilis)、質沙電氏菌marcescens) ' 嗜麥芽窄食單胞菌 (Stenotfophomonas maltophilia)、綠膝假單跑锒、洋 Μ 伯克霍涿德磨、龙潜輿允產潑磨、腦膜膿毒性黃桿菌 {Flavobacterium meningosepticum、、斯氏普羅威登斯菌 159849.doc -26-S •24- 201249786 Bacillus, Neisseria, and West genus I. In certain embodiments, an individual can be a mammal, and in some embodiments a human. Bacterial infections susceptible to the treatment of the present invention include primary infections and co-infections (co_infecti〇n) caused by bacterial species and or other infectious agents such as bacteria, viruses, parasites and fungi. . The compounds of the present invention are useful for the treatment of conditions caused by bacteria of endotoxin, and in particular by bacteria which use Gram-negative bacteria and LpxC in the biosynthesis of lipopolysaccharide (Lps) or endotoxin. The compounds of the invention are also useful in the treatment of conditions caused or exacerbated by the production of lipids and Lps or endotoxin bacteria, such as sepsis, septic shock, systemic inflammation, local inflammation, chronic obstructive pulmonary disease (c〇pD) and chronic Acute exacerbation of bronchitis (AECB). For such conditions, the treatment comprises administering a compound of the invention or a combination of the compounds of the invention, optionally together with a second agent, wherein the second agent is a second antibacterial or non-antibacterial agent. For sepsis, septic shock, systemic inflammation, local inflammation, chronic obstructive pulmonary disease (COPD), and acute exacerbation of chronic bronchitis (AECB), representative non-antibacterial agents include anti-endotoxin, including endotoxin receptor-binding antibodies, Toxin binding antibody, anti-CD14 binding protein antibody, anti-lipopolysaccharide binding protein antibody, and tyrosine kinase inhibitor. In the treatment of severe or chronic respiratory infections, the compounds of the invention may also be administered with non-antibacterial agents administered by inhalation. Representative non-antibacterial agents used in this treatment include anti-inflammatory steroids, non-steroidal anti-inflammatory agents, bronchodilators, mucolytic agents, anti-asthma therapeutics, and pulmonary surfactants. In particular, the non-antibacterial agent may be albuterol, salbuterol, budesonide, beclomethasone, dexamethasone, nedocromil, times Flumetasone, fluticasone, flunisolide, triamcinolone, ibuprofin, rofecoxib, naproxen, sene Celecoxib, nedocromil, ipratropium, metaproterenol, 11 pirbuterol, salmeterol, formoterol (formoterol), indacaterol, bronchodilator, mucolytic, calfactant, beracant, poractant alfa, sulvacin (surfaxin) or pulmozyme (also known as domase alfa). The compounds of the invention may be used alone or in combination with a second antibacterial agent to treat severe or chronic respiratory infections, including severe lung and nosocomial infections, such as infections caused by: Shame I# Mill (five "RMacier flerogewe?") , #Enterobacter cloacae, 九豫释鲁, Klebsiella pneumoniae, Klebsiella oxytoca, Proteus mirabilis, quality sand 'marscens marscens' Stenofophomonas maltophilia, green knees, single squats, artichokes Berkshire, milled dragons, dragons, sputum, sputum, septic, bacillus , Providence s. 159849.doc -26-

S 201249786 (Providencia stuartii、Sl 弗氏檸檬酸桿菌{Citrobacter ;社區肺感染，諸如由以下所引起之感染：诸减嗜血桿菌{Haemophilus Influenzae、、退伍軍人桿菌屬 {Legionella species)' Μ Μ-Μ M (Moraxella catarrhalis) ' 卡他布蘭漢氏菌{Branhamella catarrhalis、、豫择議爆、九雷伯氏菌屬及變形桿菌屬；由諸如以下之其他細菌物種所引起之感染：奈還磨屬、志貪戌磨邊（57π·^·β//α species)、沙f'氏ύ屬、幽門螺旋桿菌{Helicobacter pylori')、弧菌科 (Vibrionaceae)反博德特菌屬（BordeteUa species).，以反由布魯菌屬{Brucella 、土拉熱弗朗西絲菌 (Francisella tularensis、反I氬鼠疫耶氏桿菌所起之良染。當用於治療感染革蘭氏陰性細菌感染之個體時，本發明化合物可用以使革蘭氏陰性細菌對第二抗菌劑之作用敏感。本發明提供包括式I化合物或其立體異構體或醫藥學上可接受之鹽的新穎化合物組合，以及治療感染革蘭氏陰性細菌之個體之方法。本文所提供之新穎組合可調配成適用於本發明方法之醫藥調配物及藥劑》本發明亦提供新穎組合在製備藥劑及醫藥調配物中之用途、該組合在治療患者之細菌感染中之用途。將一種稱為棋盤分析之評估協同作用之經典方法用以預測抗菌劑之功效，且由Scribner等人（1982，Antimicrobial Agents and Chemotherapy 21(6):939-943)及 Goodman 及 159849.doc •27- 201249786S 201249786 (Providencia stuartii, Sl Citrobacter f. {Citrobacter; community lung infections, such as infections caused by: Haemophilus Influenzae, Legionella species' Μ Μ-Μ M (Moraxella catarrhalis) 'Branhamella catarrhalis, B. sinensis, sputum, N. burgdorferi and Proteus; infections caused by other bacterial species such as:志戌戌 ( (57π·^·β//α species), f ' 、, Helicobacter pylori (Helicobacter pylori), Vibrionaceae (Borcete Ua species) The invention is in accordance with the genus Brucella {Brucella, Francisella tularensis, and the genus Y. jejuni. The invention is used when treating an individual infected with a Gram-negative bacterial infection. The compounds may be used to sensitize Gram-negative bacteria to the action of the second antibacterial agent. The present invention provides novel compounds which comprise a compound of formula I or a stereoisomer or pharmaceutically acceptable salt thereof. And methods of treating an individual infected with a Gram-negative bacterium. The novel combinations provided herein can be formulated into pharmaceutical formulations and medicaments suitable for use in the methods of the invention. The present invention also provides novel combinations in the preparation of pharmaceuticals and pharmaceutical formulations. Uses, the use of this combination in the treatment of bacterial infections in patients. A classical method of evaluating synergy, called checkerboard analysis, is used to predict the efficacy of antibacterial agents, and is performed by Scribner et al. (1982, Antimicrobial Agents and Chemotherapy 21 (6). ): 939-943) and Goodman and 159849.doc •27- 201249786

Gilman(1980, The Pharmacological Basis of Therapeutics，第6版’第1097-1098頁）描述。棋盤分析涉及將抗生素個別地或以組合形式連續兩倍稀釋於培養液中，接著用待測試之微生物對其進行接種。在培育之後，測定個別使用及組合使用之各藥物之最低抑制濃度（MIC)(注意，MIC為抑制培養基中生長之最低藥物濃度協同作用由各藥物組合使用時之MIC降低來指示。拮抗作用由一或兩種藥物組合使用時之MIC提南來指示。評估協同作用之替代方法查閱 Greco# A > Pharmacological Reviews 47(2):331-285 (1995) » 其以全文引用的方式併入本文中。然而，棋盤分析中之陽性結果，亦即指示協同作用低於 MIC不一定會產生活體内協同行為。美國專利申請公開案第 2004-229955A1 號報導红黴素（erythromyCin)與 LpxC^ 制劑（即針對大腸桿菌菌株ATCC 25922之N-[(1S)-1_(胺基甲基）-2-(經胺基）-2側氧基乙基]-4-(4-{4-[({ [(3甲基苯基）曱基]胺基}乙醯基）胺基]苯基}丁-1，3二炔基）苯甲醯胺）之間的強烈協同作用。國際pct專利申請案第PCT/US2〇1〇/ 33910號證明紅黴素及多種Lpxc抑制劑之組合顯示無活體内協同作用。Gilman (1980, The Pharmacological Basis of Therapeutics, 6th Edition, pp. 1097-1098) is described. Chessboard analysis involves the sequential dilution of antibiotics in culture medium, either individually or in combination, followed by inoculation with the microorganism to be tested. After incubation, the minimum inhibitory concentration (MIC) of each drug used individually and in combination was determined (note that the MIC is the lowest drug concentration in the inhibition medium and the synergy is indicated by the decrease in MIC when each drug combination is used. Antagonism is indicated by One or two combinations of drugs are used to indicate the MIC. An alternative method for assessing synergy is reviewed in Greco# A > Pharmacological Reviews 47(2): 331-285 (1995) » It is incorporated herein by reference in its entirety. However, the positive result in the checkerboard analysis, that is, the indication that the synergy is lower than the MIC, does not necessarily lead to in vivo synergistic behavior. U.S. Patent Application Publication No. 2004-229955A1 reports erythromycyl (Cinomyl Cin) and LpxC^ preparations ( That is, N-[(1S)-1_(aminomethyl)-2-(amino-amino)-2-oxyethyl]-4-(4-{4-[({) for E. coli strain ATCC 25922 Strong synergy between [(3methylphenyl)indolyl]amino}ethyl)amino]phenyl}butyl-1,3diynyl)benzamide. International pct patent application PCT/US2〇1〇/ 33910 demonstrates the combination of erythromycin and various Lpxc inhibitors There is no synergy within the living body.

LpxC ’即革蘭氏陰性細菌中之必需基因，編碼酶尿苦二磷酸-3-〇-(R-羥基癸醯基）_N-乙醯葡萄糖胺去乙醯基酶。此酶催化生物合成脂質A(即作為所有革蘭氏陰性細菌之必需組分之脂多醣的脂質部分）中之早期關鍵步驟分。高於 MIC ’則預期LpxC抑制劑會破壞外膜，因此允許其他抗菌 159849.doc -28- 201249786 化合物滲透外膜。一旦此等藥劑已滲透外膜，其即可影響周質目標，如同用萬古黴素之情况一般，或其可接著擴散穿過内膜以與諸如核糖體（紅黴素）或核糖核酸聚合酶（利福平）之細胞内目標相互作用。在無LpxC抑制劑存在下，諸如萬古黴素之藥劑接近其目標之能力因外膜而大大降低。因此，不受理論束缚，咸信可能下伏於所觀測到之協同作用之生物化學機制為外膜對與11^(：抑制劑組合時的諸如萬古黴素之藥劑之滲透性增強。在一實施例中，與式Ϊ化合物或其立體異構體或醫藥學上可接$之鹽組合使用之第二抗菌劑為萬古徽f '利奈。坐胺、阿奇黴素、亞胺培南、替考拉甯、達托黴素、克林達黴素、利福平、頭孢噻肟、慶大黴素、新生黴素或特拉萬星。在一個該實施例中，第二抗菌劑為萬古黴素、替考拉寧、利福平、阿奇黴素、特拉萬星或新生黴素。在一個該實施例中，第二抗菌劑為萬古黴素或利福平。在本發明之某些實施例中，投與低於治療劑量之第二抗菌劑及/或式I 化合物或其立體異構體或醫藥學上可接受之鹽，其中低於治療劑量為單獨投與時不足以治療細菌感染之劑量。本發明之醫藥組合物包含式〗化合物或其立體異構體或醫藥學上可接受之鹽連同一或多種醫藥學上可接受之載劑或稀釋劑一起調配。如本文所用，術語「醫藥學上可接受之載劑」意謂無毒性之惰性固體、半固體或液體填充劑、稀釋劑、囊封材料或任何類型之調配助劑。可充當醫藥學上可接受之載劑的材料之一些實例為糖，諸如乳糖、葡萄 I59849.doc -29- 201249786 糖及蔗糖；澱粉，諸如玉米澱粉及馬鈴薯澱粉；纖維素及其衍生物，諸如羧曱基纖維素鈉、乙基纖維素及乙酸纖維素；粉狀黃蓍；麥芽；明膠；滑石；賦形劑，諸如諸如可可脂及栓劑蠟；油，諸如花生油、棉籽油；紅花子油；芝麻油；撖欖油；玉米油及大豆油；二醇，諸如丙二醇；醋’諸如油酸乙酯及月桂酸乙酯；瓊脂；緩衝劑，諸如氣氧化鎮及氫氧化鋁；海藻酸；無熱原質水；等張生理食鹽水；林格氏溶液（Ringer's solution);乙醇及磷酸鹽緩衝溶液以及其他無毒相容性潤滑劑（諸如月桂基硫酸鈉及硬脂酸鎂）以及著色劑、釋放劑、塗佈劑、甜味劑、調味劑及芳香劑、防腐劑及抗氧化劑根據調配者之判斷亦可存在於組合物中。可將本發明之醫藥組合物經口、經直腸、非經腸（如藉由靜脈内、肌肉内或皮下注射）、腦池内、*** 内、腹膜内、局部（如藉由散劑、軟膏或滴劑）、經頰或以經口或經鼻喷霧劑形式，或用於吸入之液體氣溶膠或乾粉調配物形式投與人類及其他動物。經口投藥之液體劑型包括醫藥學上可接受之乳液、微乳液、溶液、懸浮液、糖漿及酏劑。除活性化合物外，液體劑型亦可含有通常用於此項技術中之惰性稀釋劑，例如水或其他溶劑、助溶劑及乳化劑’諸如乙醇、異丙醇、碳酸乙酯、乙酸乙酯、苯甲醇、苯甲酸苯甲酯、丙二醇、丨，3_ 丁二醇、二曱基曱醯胺、油（特別是棉籽油、落花生油、玉米油、胚芽油、橄欖油、蓖麻油及芝麻油）、甘油、四氫糠醇、聚乙二醇及脫水山梨糖醇脂肪酸醋，及其濃合 159849.doc -30· a 201249786 物。除惰性稀釋劑外’ Π服組合物亦可包括佐劑，諸如潤濕劑、乳化劑及懸浮劑、甜味劑、調味劑及芳香劑。4 可根據已知技術使用適合分散劑或潤濕劑及懸浮劑來調配可注射製劑，例如無菌可注射水性或油性懸浮液。無菌可主射製劑亦可為於無毒、非腸胃可接受之稀釋劑或溶劑中之無菌可注射溶液、懸浮液或乳液，例如於丁二醇中之溶液。在可接受之媒劑及溶劑中，可採用水、林格氏溶液、1%利乡卡因（lidocaine)、u s p及等張氯化納溶液。此外，無菌不揮發性油習用作溶劑或懸浮介質。為本發明之目#’可採用任何溫和不揮發性油，包括合成之單酸甘油醋及二酸甘油醋。另夕卜，在可注射劑之製備中使用脂肪酸，諸如油酸。 J注射調配物可例如，，，土《渊困戰筲過濾w μ嗯承飒菌，或藉由在使用前併有呈可溶解或分散於無菌水或其他I菌可注射介質中之無菌固體組合物形式之滅菌劑來滅菌。為延長藥物作用，通常需要減緩來自皮下或肌肉内注射之藥物的吸收。此舉可藉由使用具有不良水溶性之晶體或非晶物質之液體料液來達成1物吸收速率則視其溶解速率而^ ’而溶解速率又可視晶體大小及晶形而定。或者’非經腸投藥之藥物形式之延遲吸收可藉由將藥物溶於，懸浮於油媒劑中來達成。藉由在可生物降解之聚合物 (諸如聚丙父醋-聚乙交醋）中形成藥物之微膠囊基質來製得 β、射儲槽形式。視藥物與聚合物之比率及所用特定聚合物之性質而定，可控制藥物釋放速率。其他可生物降解： 159849.doc •31· 201249786 聚合物的實例包括聚（原酸酯）及聚（酐）。亦可藉由將藥物覆埋入與身體組織相容之脂質體或微乳液中來製備儲槽式可注射調配物。用於經直腸或經***投藥之組合物較佳為栓劑，該等栓劑可藉由將本發明化合物與適合非刺激性賦形劑或載劑 (如可可脂、聚乙二醇或栓劑蠟）混合來製備，該等賦形劑或載劑在環境溫度下為固體但在體溫下為液體，因此在直腸或***空腔中融化並釋放該活性化合物。用於經口投藥之固體劑型包括膠囊、錠劑、丸劑、散劑及顆粒劑。在該等固體劑型中，活性化合物與至少一種諸如檸檬酸鈉或磷酸二鈣之醫藥學上可接受之惰性賦形劑或載劑及/或以下混合：a)填充劑或增量劑，諸如澱粉、乳糖、蔗糖、葡萄糖、甘露糖醇及矽酸；b)黏合劑，諸如緩甲基纖維素、海藻酸鹽、明膠、聚乙烯吼咯啶酮、蔗糖及 ***膠；c)保濕劑，諸如丙三醇；d)崩解劑，諸如瓊脂、碳酸鈣、馬鈴薯或木薯澱粉、海藻酸、某些矽酸鹽及碳酸納；e)溶液阻滯劑，諸如石蠟；f)吸收促進劑，諸如四級銨化合物；g)濕潤劑，諸如乙醇（acetyl ale〇hol)及單硬脂酸甘油酯；h)吸附劑，諸如高嶺土（kaolin)及膨潤土；及i)潤滑劑’諸如滑石、硬脂酸鈣、硬脂酸鎂、固體聚乙二醇、月桂基硫酸鈉及其混合物。在膠囊、錠劑及丸劑之情況下’劑型亦可包含緩衝劑。類似類型之固體組合物亦可用作使用諸如乳糖 (lactose ; milk sugar)以及高分子量聚乙二醇及其類似物之 159849.doc •32-LpxC' is an essential gene in Gram-negative bacteria, encoding the enzyme urinary diphosphate 3-〇-(R-hydroxyindenyl)_N-acetylglucosamine deacetylase. This enzyme catalyzes an early critical step in the biosynthesis of lipid A, the lipid portion of lipopolysaccharide, which is a required component of all Gram-negative bacteria. Above the MIC', LpxC inhibitors are expected to damage the outer membrane, thus allowing other antimicrobial compounds to penetrate the outer membrane. Once these agents have penetrated the outer membrane, they can affect the periplasmic target, as is the case with vancomycin, or it can then diffuse through the inner membrane to interact with, for example, ribosomes (erythromycin) or ribonucleic acid polymerase. Intracellular target interactions (rifampicin). In the absence of LpxC inhibitors, the ability of agents such as vancomycin to approach its target is greatly reduced by the outer membrane. Therefore, without being bound by theory, the biochemical mechanism underlying the observed synergy is the enhanced permeability of the outer membrane pair to the agent such as vancomycin when the inhibitor is combined. In the examples, the second antibacterial agent used in combination with the guanidine compound or a stereoisomer thereof or a pharmaceutically acceptable salt of the genus is the genus F' Lina. Osmotic amine, azithromycin, imipenem, teicoplana Ning, daptomycin, clindamycin, rifampicin, cefotaxime, gentamicin, novomycin or telavancin. In one such embodiment, the second antibacterial agent is vancomycin , teicoplanin, rifampicin, azithromycin, telavancin or novomycin. In one such embodiment, the second antimicrobial agent is vancomycin or rifampicin. In certain embodiments of the invention Administering a second antibacterial agent and/or a compound of formula I, or a stereoisomer or pharmaceutically acceptable salt thereof, below the therapeutic dose, wherein the therapeutic dose below the therapeutic dose is insufficient to treat the bacterial infection. The pharmaceutical composition of the present invention comprises a compound of the formula or a stereo Isomers or pharmaceutically acceptable salts are formulated with one or more pharmaceutically acceptable carriers or diluents. As used herein, the term "pharmaceutically acceptable carrier" means non-toxic inertness. Solid, semi-solid or liquid fillers, diluents, encapsulating materials or any type of formulation aid. Some examples of materials that can serve as pharmaceutically acceptable carriers are sugars such as lactose, grape I59849.doc -29 - 201249786 Sugar and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered scutellaria; malt; gelatin; talc Excipients such as, for example, cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; eucalyptus oil; corn oil and soybean oil; glycols such as propylene glycol; vinegar such as ethyl oleate And lauric acid ethyl ester; agar; buffer, such as gas oxidation town and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; Phosphate buffer solution and other non-toxic compatible lubricants (such as sodium lauryl sulfate and magnesium stearate) as well as coloring agents, release agents, coating agents, sweeteners, flavoring and fragrances, preservatives and antioxidants It may also be present in the composition according to the judgment of the formulator. The pharmaceutical composition of the present invention may be administered orally, rectally, parenterally (e.g., by intravenous, intramuscular or subcutaneous injection), intracranial, intravaginal, Humans and other animals are administered intraperitoneally, topically (e.g., by powder, ointment or drops), buccally or in the form of an oral or nasal spray, or in the form of a liquid aerosol or dry powder formulation for inhalation. The liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, the liquid dosage form may contain inert diluents which are conventionally used in the art. For example, water or other solvents, co-solvents and emulsifiers 'such as ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, hydrazine, 3-butanediol, dimercapto Guanamine, oil (especially cottonseed oil, groundnut oil, corn oil, germ oil, olive oil, castor oil and sesame oil), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acid vinegar, and their rich 159849.doc -30· a 201249786. In addition to the inert diluent, the composition may also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, and perfuming agents. Injectable preparations, e.g., sterile injectable aqueous or oily suspensions, may be employed in accordance with known techniques using suitable dispersing or wetting agents and suspending agents. The sterile, active preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic, parenterally acceptable diluent or solvent, such as a solution in butanediol. Among the acceptable vehicles and solvents, water, Ringer's solution, 1% lidocaine, u s p and isotonic sodium chloride solution can be used. In addition, sterile, fixed oils are conventionally employed as a solvent or suspension medium. Any of the mild, fixed oils may be employed for the purpose of the present invention, including synthetic monoglycerides and diglycerides. In addition, fatty acids such as oleic acid are used in the preparation of injectables. The J-injection formulation can be, for example, a soil, a dampness filter, or a sterile solid that is soluble or dispersible in sterile water or other injectable medium prior to use. The sterilizing agent in the form of a composition is sterilized. In order to prolong the action of the drug, it is usually necessary to slow the absorption of the drug from subcutaneous or intramuscular injection. This can be achieved by using a liquid feed liquid having a poorly water-soluble crystal or amorphous substance to achieve a rate of absorption of the substance, depending on the dissolution rate, and the dissolution rate depending on the crystal size and crystal form. Alternatively, delayed absorption of the parenterally administered drug form can be accomplished by dissolving the drug in suspension in an oil vehicle. The β, sump form is prepared by forming a microcapsule matrix of the drug in a biodegradable polymer such as polypropylene vinegar-polyacetate. The drug release rate can be controlled depending on the ratio of drug to polymer and the nature of the particular polymer used. Other biodegradable: 159849.doc • 31· 201249786 Examples of polymers include poly(orthoesters) and poly(anhydrides). A reservoir-type injectable formulation can also be prepared by burying the drug in a liposome or microemulsion that is compatible with body tissues. Compositions for rectal or vaginal administration are preferably suppositories which can be prepared by combining a compound of the invention with a suitable non-irritating excipient or carrier (such as cocoa butter, polyethylene glycol or suppository wax) Prepared by mixing, the excipients or carriers are solid at ambient temperature but liquid at body temperature, thus melting and releasing the active compound in the rectal or vaginal cavity. Solid dosage forms for oral administration include capsules, lozenges, pills, powders, and granules. In such solid dosage forms, the active compound is mixed with at least one pharmaceutically acceptable inert excipient or carrier such as sodium citrate or dicalcium phosphate and/or below: a) a filler or extender, such as Starch, lactose, sucrose, glucose, mannitol and citric acid; b) binders such as slow methylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose and gum arabic; c) humectants, Such as glycerol; d) disintegrants such as agar, calcium carbonate, potato or tapioca starch, alginic acid, certain citrates and sodium carbonate; e) solution blockers such as paraffin; f) absorption enhancers, Such as quaternary ammonium compounds; g) humectants such as ethanol (acetyl ale〇hol) and glyceryl monostearate; h) adsorbents such as kaolin and bentonite; and i) lubricants such as talc, hard Calcium citrate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, lozenges and pills, the dosage form may also contain a buffer. Solid compositions of a similar type may also be employed as the use of, for example, lactose; milk sugar as well as high molecular weight polyethylene glycols and the like, 159849.doc • 32-

S 201249786 賦形劑的填充軟明膠膠囊及硬明膠膠囊中的填充劑。可以使用包衣及外殼（諸如腸溶衣及醫藥調配技術中熟知之其他包衣）來製備錠劑、糖衣藥丸、膠囊、丸劑及顆粒劑之固體劑型。其可視情況含有乳濁劑，且亦可為僅在或優先在腸道之某一部分視情況以延遲方式釋放活性成分之組合物。可使用之包埋組合物之實例包括聚合物質及蠟。類似類型之固體組合物亦可用作使用諸如乳糖以及高分子量聚乙二醇及其類似物之賦形劑的填充軟明膠膠囊及硬明膠膠囊中的填充劑。抗菌化合物亦可與一種或多種如上所述之賦形劑一起呈微膠囊形式。可以使用包衣及外殼（諸如腸溶衣、釋放控制包衣及醫藥調配技術中熟知之其他包衣）來製備錠劑、糖衣藥丸、膠囊、丸劑及顆粒劑之固體劑型。在該等固體劑型中，活性化合物可與至少一種惰性稀釋劑（諸如蔗糖、乳糖或澱粉）混合。如在正常實踐中，該等劑型亦可包含除惰性稀釋劑外之其他物質，例如製錠潤滑劑及其他製錠助劑，鈕如硬脂.酸鎂及微晶纖維素。在膠囊、錠劑及丸劑之情況下，該等劑型亦可包含緩衝劑。其可視情況含有礼濁劑，且亦可為僅在或優先在腸道某一部分視情況以延遲方式釋放活性成分之組合物。可使用之包埋組合物之實例包括聚合物質及蠟。用於局部投與或級皮投與本發明化合物之劑型包括軟膏、糊劑、乳膏、洗劑、凝膠、散劑、溶液、噴霧劑、吸 159849.doc •33- 201249786 入d或貼）i °在無菌條件下，將活性組分與醫藥學上可接受之載劑及任何所需之防腐劑或可能需要之緩衝劑混合。眼用調配4耳劑及其類似物亦涵蓋於本發明之範缚内。除本發明之活性化合物外’軟膏、糊劑、乳膏及凝膠亦可含有賦形劑’諸如動物及植物脂肪、油、蠟、石蠟、澱粉、黃蓍、纖維素衍生物、m、聚♦氧、膨潤土、石夕酸、滑石及氧化鋅或其混合物。本發明之組合物亦可經調配為液體氣溶膠或可吸入乾粉來傳遞。液體氣溶膠調配物可主要喷霧成可傳遞至末端及呼吸性細支氣管（其中細菌存在於患有諸如慢性支氣管炎及肺炎之支氣管感染的患者中）之粒度。病原菌通常存在於整個氣管直至支氣管、細支氣管及肺實質，尤其在末端及呼吸性細支.氣管中。在感染惡化期間，細菌亦可存在於肺泡中β液體氣溶膠及可吸入乾粉調配物較佳在整個支氣管内樹中傳遞至末端細支氣管且最終傳遞至實質組織。本發明之氣溶膠化調配物可使用諸如射流、振動多孔板或超音波喷霧器之氣溶膠形成器件（較佳經選擇為可形成質量中值平均直徑主要在1 μιη至5 μπι之間的氣溶膠粒子）進行傳遞。此外，調配物較佳具有平衡容積莫耳滲透濃度離子強度及氣化物.濃度，及能將有效劑量之本發明化合物傳遞至感染部位之最小可氣溶膠化體積。另外，氣溶膠化調配物較佳不會消極地削弱氣管之功能性且不會引起不合乎需要之副作用。 159849.doc -34-S 201249786 Filled soft gelatin capsules of excipients and fillers in hard gelatin capsules. Solid dosage forms of lozenges, dragees, capsules, pills, and granules can be prepared using coatings and shells, such as enteric coatings and other coatings well known in the art. It may optionally contain an opacifying agent and may be a composition which will release the active ingredient in a delayed manner, as appropriate or preferentially in a portion of the intestinal tract. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type may also be employed as fillers in filled soft gelatin capsules and hard gelatin capsules using such excipients as lactose and high molecular weight polyethylene glycols and the like. The antibacterial compound may also be in microencapsulated form with one or more excipients as described above. Solid dosage forms of lozenges, dragees, capsules, pills, and granules can be prepared using coatings and shells such as enteric coatings, release control coatings, and other coatings well known in the art. In such solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose or starch. In normal practice, such dosage forms may also contain other materials than inert diluents, such as tableting lubricants and other tableting aids, such as stearin, magnesium sulphate and microcrystalline cellulose. In the case of capsules, lozenges and pills, such dosage forms may also contain buffering agents. It may optionally contain a turbidity agent and may also be a composition which will release the active ingredient in a delayed manner, as appropriate or preferentially in a certain portion of the intestinal tract. Examples of embedding compositions that can be used include polymeric materials and waxes. Dosage forms for topical administration or graded administration of the compounds of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, 159849.doc • 33- 201249786 into d or paste) I ° Under sterile conditions, the active component is mixed with apharmaceutically acceptable carrier and any required preservatives or buffers which may be required. Ophthalmic formulation of 4 ear and its analogs are also encompassed within the scope of the invention. In addition to the active compounds of the invention, ointments, pastes, creams and gels may also contain excipients such as animal and vegetable fats, oils, waxes, paraffin, starch, xanthine, cellulose derivatives, m, poly ♦ Oxygen, bentonite, aspartic acid, talc, and zinc oxide or mixtures thereof. The compositions of the present invention may also be formulated as a liquid aerosol or an inhalable dry powder for delivery. The liquid aerosol formulation can be primarily sprayed into a particle size that can be delivered to the terminal and respiratory bronchioles where the bacteria are present in a patient suffering from a bronchial infection such as chronic bronchitis and pneumonia. Pathogens usually exist throughout the trachea up to the bronchi, bronchioles and lung parenchyma, especially in the distal and respiratory branches. During the deterioration of the infection, the bacteria may also be present in the alveoli. The beta liquid aerosol and the respirable dry powder formulation are preferably delivered to the terminal bronchioles throughout the bronchial tree and ultimately to the parenchyma. The aerosolized formulation of the present invention may employ an aerosol-forming device such as a jet, a vibrating porous plate or an ultrasonic nebulizer (preferably selected to form a mass median average diameter of between 1 μm and 5 μm). Aerosol particles) are delivered. In addition, the formulation preferably has a balance volume osmolality, ionic strength and vapor concentration, and a minimum aerosolizable volume capable of delivering an effective amount of a compound of the invention to the site of infection. In addition, the aerosolized formulation preferably does not negatively impair the functionality of the trachea and does not cause undesirable side effects. 159849.doc -34-

S 201249786 適用於投與本發明氣溶膠調配物之氣溶膠化器件包括例如能夠將本發明調配物喷霧成粒度主要在1 pm至5 pm尺寸範圍内之氣溶膠之射流、振動多孔板、超音波噴霧器及受激勵乾粉吸入器。在本申請案中主要意謂所有所產生之氣溶膠粒子之至少70%，但較佳90%以上在1 μιη至5 μιη之範圍内。射流喷霧器藉由氣壓工作以將液體溶液打碎成氣溶膠小滴。振動多孔板喷霧器藉由使用由快速振動多孔板所產生之音波真空以將溶劑小滴擠壓穿過多孔板來工作。超音波喷霧器藉由將液體剪切成小氣溶膠小滴之壓電晶體來工作。可用多種適合器件，包括例如AeroNeb及AeroDose 振動多孔板喷霧器（AeroGen，Inc.，Sunnyvale，Calif.) ' Sidestream7 喷霧器（Medic-Aid Ltd., West Sussex, England)、Pari LC7 及 Pari LC Star7 射流喷霧器（Pari Respiratory Equipment, Inc., Richmond, Va.)及 Aerosonic (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany)及 UltraAire7(Omron Healthcare, Inc., Vernon Hills, 111.)超音波喷霧器。本發明化合物亦可經調配用作除本發明化合物外亦可含有諸如以下之賦形劑的局部散劑及喷霧劑：乳糖、滑石、矽酸、氫氧化鋁、矽酸鈣及聚醯胺粉末或此等物質之混合物。喷霧劑可另外含有習用推進劑，諸如氣氟烴。經皮貼片具有提供化合物至身體之控制傳遞之附加優勢。該等劑型可藉由於適合介質中溶解或分配化合物來製得。亦可使用吸收增強劑來增加化合物之透皮通量。可藉 159849.doc -35- 201249786 由提供速率控制膜或將化合物分散於聚合物基質或凝膠來控制速率。根據本發明之治療方法，藉由向患者(諸如人類或低等哺乳動物）投與治療有效量之式恤合物或其立體醫藥學上可接受之鹽、用獲得所需結果所必需之量及時間來治療或預防患者之細菌感染。本發明化合物之「治療有效量」意谓足以用適用於任何醫療之合理收益/風險比治療細菌感染之化合物的量。然而，應瞭解，本發明之化合物及組合物之總日用量將由主治醫師在正確醫學判斷範：内來決^。任何特定患者之特定治療有效劑量均將視多種因素而定’該等因素包括所治療之病症及該病症之嚴重度；所用特定化合物之活性；所用特定組合物；患者之年 7體重般健康狀況、性別及腊食；投藥時間、投藥途徑及所用特定化合物之***速率；治療持續時間，與所用特定化合物組合或同時使用之藥物；及醫學技術中熟知之類似因素。以單次劑量或分次劑量形式投與人類或其他哺乳動物之本發明化合物之總日劑量總計可為例如每公斤體重〇〇1 mg至200 mg，或更通常為每公斤體重〇1 ^^至兄。在某些實施例中，投與人類或其W動物之總日劑量為每公斤體重1.0 111§至100 mg或每公斤體重5 〇 ^^至25 。單次劑量組合物可含有組成曰劑量之量或其約量。一般而言，本發明之治療方案包含每天以單次劑量或多次劑量形式向需要該治療之患者投與約10 mg至約15 g本發明化合 159849.doc a •36· 201249786 物’更通常為100 mg至5 g，且甚至更通常每天以單次或多次劑量形式投與250 mg至1 g。調配方法在此項技術中熟知且揭示於例如S 201249786 Aerosolized devices suitable for administration to the aerosol formulations of the present invention include, for example, aerosols capable of spraying the formulations of the present invention into aerosols having a particle size predominantly in the range of 1 pm to 5 pm, vibrating perforated plates, super Sonic sprayer and activated dry powder inhaler. In the present application it is meant primarily that at least 70%, but preferably more than 90%, of all of the aerosol particles produced are in the range of 1 μηη to 5 μιη. The jet nebulizer operates by air pressure to break up the liquid solution into aerosol droplets. The vibrating perforated plate sprayer works by using a sonic vacuum generated by a rapidly vibrating porous plate to squeeze solvent droplets through the perforated plate. Ultrasonic nebulizers work by shearing a liquid into a small aerosol droplet of a piezoelectric crystal. A wide range of suitable devices are available, including, for example, AeroNeb and AeroDose Vibrating Perforated Plate Sprayers (AeroGen, Inc., Sunnyvale, Calif.) 'Sidestream7 Sprayers (Medic-Aid Ltd., West Sussex, England), Pari LC7 and Pari LC Star7 Jet Sprayer (Pari Respiratory Equipment, Inc., Richmond, Va.) and Aerosonic (DeVilbiss Medizinische Produkte (Deutschland) GmbH, Heiden, Germany) and UltraAire 7 (Omron Healthcare, Inc., Vernon Hills, 111.) Ultrasonic sprayer. The compounds of the present invention may also be formulated for use as a topical powder and spray in addition to the compound of the present invention, such as lactose, talc, citric acid, aluminum hydroxide, calcium citrate and polyamido powder. Or a mixture of such substances. Sprays may additionally contain customary propellants, such as fluorocarbons. Transdermal patches have the added advantage of providing controlled delivery of the compound to the body. Such dosage forms can be prepared by dissolving or dispensing the compound in a suitable medium. Absorption enhancers can also be used to increase the transdermal flux of the compound. The rate can be controlled by providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel by 159849.doc -35- 201249786. According to the method of the present invention, a therapeutically effective amount of a chelating composition or a pharmaceutically acceptable salt thereof is administered to a patient (such as a human or a lower mammal) in an amount necessary to obtain a desired result. And time to treat or prevent bacterial infections in patients. The "therapeutically effective amount" of a compound of the invention means an amount sufficient to treat a bacterial infection with a reasonable benefit/risk ratio for any medical treatment. It will be understood, however, that the total daily usage of the compounds and compositions of the present invention will be determined by the attending physician in the correct medical judgment. The particular therapeutically effective dose for any particular patient will depend on a number of factors' such factors as the condition being treated and the severity of the condition; the activity of the particular compound employed; the particular composition employed; , sex and bacon; time of administration, route of administration and rate of excretion of the particular compound used; duration of treatment, combination with or in combination with the particular compound employed; and similar factors well known in the medical arts. The total daily dose of a compound of the invention administered to a human or other mammal in a single or divided dose may total, for example, from 1 mg to 200 mg per kilogram of body weight, or more typically 1 ^^ per kilogram of body weight. To the brother. In certain embodiments, the total daily dose administered to a human or a W animal is 1.0 111 § to 100 mg per kilogram of body weight or 5 〇 ^^ to 25 per kilogram of body weight. A single dose of the composition may contain the amount of the sputum dose or an amount thereof. In general, the treatment regimen of the present invention comprises administering to a patient in need of such treatment about 10 mg to about 15 g of the present invention in a single dose or in multiple doses per day. 159849.doc a • 36· 201249786 From 100 mg to 5 g, and even more usually, 250 mg to 1 g is administered in a single or multiple doses per day. Methods of formulation are well known in the art and are disclosed, for example, in

Remington:Remington:

The Science and Practice of Pharmacy, Mack Publishing Company，Easton，Pa·，第19版（1995)中。用於本發明之醫藥組合物可呈無菌、非熱解液體溶液或懸浮液、包覆包衣之膠囊、栓劑、凍乾散劑、經皮貼片形式或此項技術中已知之其他形式。如本申請案中所用’「套組」包括含有醫藥組合物之容器，且亦可包括分隔容器，諸如分隔瓶或分隔箔包裝。該容器可呈在此項技術中已知之任何習知形狀或形式其係由例如以下之醫藥學上可接受之材料製成：紙或紙板盒、玻璃或塑膠瓶或罐、可重密封袋（例如保存錠劑之「再充填物」以置於不同容器中），或具有根據治療時程自包裝中按壓出之個別劑量的泡殼包裝。所用容器可視所涉及之精確劑型而定，例如習知之紙板盒通常不會用以保存液體懸浮液。可行的是可將—個以上之容器—起用於單一包農The Science and Practice of Pharmacy, Mack Publishing Company, Easton, Pa., 19th edition (1995). The pharmaceutical compositions for use in the present invention may be in the form of a sterile, non-pyrolyzed liquid solution or suspension, a coated capsule, a suppository, a lyophilized powder, a transdermal patch or other forms known in the art. The "set" as used in this application includes a container containing a pharmaceutical composition, and may also include a separate container, such as a separate bottle or a separate foil package. The container may be in any conventional shape or form known in the art and is made of, for example, the following pharmaceutically acceptable materials: paper or cardboard boxes, glass or plastic bottles or cans, resealable bags ( For example, a "refill" of a lozenge is stored for placement in a different container, or a blister pack having individual doses that are pressed from the package according to the duration of the treatment. The container used may depend on the precise dosage form involved. For example, conventional cardboard boxes are generally not used to hold liquid suspensions. It is feasible to use more than one container for a single package

中以出售單一劑型。舉例而言，錠劑可容納於瓶中，而該瓶又容納於盒子内0 X 該套組之實例為所謂泡殼包裝。泡殼包裝在包裝行業中已熟知且正廣泛用於包裝醫藥單位劑型(錠劑、膠囊及立類似劑型卜泡殼包裝通常由一片相對較：透明之塑膠材料箔覆M細占..„ 自覆蓋組成。在包裝過程期間，於塑膠箔令形成凹座。該等凹座具有待包裝之個別錢劑或膠囊之尺 159849.doc -37- 201249786 寸及形狀或可具有容納多個待包裝之錠劑及/或膠囊之尺寸及形狀。接下來，將錠劑或膠囊相應地置於該等凹座中，且在該落與凹座形成方向相反的面上與塑膠猪相抵來密封該片相對較硬之材料。因此，必要時將錠劑或膠囊個別密封或共同密封於塑膠箔與該片之間的凹座中。較佳地，該片之強度使得錠劑或膠囊可藉由手工施壓於凹座上、藉此在該片中之凹座位置處形成開口而自泡殼包裝中移除。接著可經由該開口移除錠劑或膠囊。可能需要提供-種書面記憶輔助物’其中該書面記憶輔助物之類型為含有用於醫師、藥劑師或其他健康照護提供者或個體之資訊及/或說明，例如呈以下形式：緊接於錠劑或膠囊之數字，藉此該等數字與應攝取所指定之鍵劑或膠囊之療程天數對應，或含有相同類型資訊之卡片。該記憶輔助物之另一實例為印刷於卡片上之行事層，例如：；「第一周、週一、週二…」等…「第二週、週一、週二…」等。記憶輔助物之其他變化將顯而易知。「曰劑量」可為待於既定日月艮用之單一錠劑或膠囊或若干旋劑或膠囊。當套組含有各別組合物時，套組之—或多種組合物之日劑量可由一錠劑或膠囊組《，而套組之其他一或多種組合物之日劑量可由若干錠劑或膠囊組成。套組之另一特定實施例為一種經設計按其預定用途一次 -者分配日劑量的分配器。該分配器較佳裝備有記憶輔助物’以便進-步有助於療程内之順應性。該記憶辅助物之一實例為指示已分配之曰劑量數目之機械計數器。該記憶 159849.doc 00 -38 · a 201249786 辅助物之另-實例為電池供電微晶片記憶體與以下耗接·· 例如讀出最後曰劑量之服用曰期及/或提醒何時服用下一劑的液晶讀出器或聲訊提醒信號。本發明套組除本發明化合物外亦可包括一或多種其他醫藥學活性化合物。舉例而言’其他化合物為第二抗菌劑。其他化合物可以與本發明化合物相同之劑型或以不同劑型投與。同樣地’其他化合物可與本發明化合物同時或不同時投與^ 本發明化合物之組合物亦可與具有類似抗菌範圍之其他已知抗菌劑組合使用以（1)增強由此化合物之抗菌範圍覆蓋之嚴重革蘭氏陰性感染之治療，或（2)增加懷疑有多種生物體之嚴重感染中之覆蓋，其中除此化合物之外亦可能需要具有不同抗菌範圍之另一藥劑β可能藥劑包括胺基醣苷、月徽素（penicillin)、頭孢菌素（cephai0Sp0rin)、ι喧諾酮 (fluoroquinolone)、巨環内酯（macr〇Ude)、糖肽、脂肽及噁唑啶酮之成員。治療可涉及投與具有本發明化合物與第二抗菌化合物之組合物或投與本發明化合物之一種化合物，之後或之前投與第二抗菌劑。上文可藉由參考以下實例更好地瞭解，該等實例為說明而呈現且不限制本發明概念之範疇。實例參考以下實例，本發明化合物藉由高效液相層析法 (HPLC)使用具有2690分離模組之Waters Millenium層析系統（Milford，Mass)表徵。分析管柱為Alltima C-18逆相管 159849.doc •39- 201249786 柱，4.6x250 mm，來自 Alltech(Deerfield，111.)。使用通常以5%乙腈/95%水起始且於40分鐘之期間内進展至ι〇〇%乙腈的梯度溶離。所有溶劑均含有〇 · 1 °/。三氟乙酸（TFA)。藉由在220 nm或254 nm下之紫外光（UV)吸收來偵測化合物。在一些情況下’藉由薄層層析（TLC)使用玻璃或塑膠襯底之石夕膠板（諸如Baker-Flex石夕膠1 B2-F可撓性薄片）來評估純度。TLC結果易於在紫外光下或藉由採用熟知之峨蒸氣及其他各種染色技術來視覺偵測。質譜分析經以下兩個LCMS儀中之一者進行：waters系統（Alliance HT HPLC 及Micromass ZQ質譜儀；管柱：In order to sell a single dosage form. For example, the tablet can be contained in a bottle, which in turn is contained in a box. 0 X An example of the set is a so-called blister pack. The blister packaging is well known in the packaging industry and is widely used in the packaging of pharmaceutical unit dosage forms (tablets, capsules and similar dosage forms. The blister packaging is usually made up of a relatively thin piece: transparent plastic material foil covering M fine.. „ Covering composition. During the packaging process, the plastic foil is formed into a recess. The recesses have the dimensions of the individual money or capsules to be packaged 159849.doc -37 - 201249786 and may have a plurality of packages to be packaged. The size and shape of the tablet and/or capsule. Next, the tablet or capsule is placed in the recesses correspondingly, and the plastic pig is placed against the surface opposite to the direction in which the recess is formed to seal the sheet. A relatively hard material. Therefore, if necessary, the tablet or capsule is individually sealed or co-sealed in a recess between the plastic foil and the sheet. Preferably, the strength of the sheet allows the tablet or capsule to be manually processed. Applying pressure to the recess, thereby removing the opening from the blister pack at the recess location in the sheet. The lozenge or capsule can then be removed via the opening. It may be desirable to provide a written memory aid 'Where the written Recall that the type of accessory is information and/or instructions for use by a physician, pharmacist, or other health care provider or individual, such as in the form of a number immediately following a lozenge or capsule, whereby such numbers and The number of days of treatment for the specified key or capsule, or a card containing the same type of information. Another example of the memory aid is the actuating layer printed on the card, for example: "First week, Monday, week Second..."etc..."Second week, Monday, Tuesday..." etc. Other changes in memory aids will be readily apparent. "曰 dose" can be a single lozenge or capsule to be used for a given day or month. Or a plurality of spinners or capsules. When the kit contains the respective compositions, the daily dose of the kit or compositions may be from a lozenge or capsule group, and the daily dose of the other composition or compositions of the kit It may be comprised of a plurality of tablets or capsules. Another specific embodiment of the kit is a dispenser designed to dispense a daily dose once for its intended use. The dispenser is preferably equipped with a memory aid' for further step-by-step Help with treatment Compliance. An example of such a memory aid is a mechanical counter indicating the number of doses dispensed. The memory 159849.doc 00 -38 · a 201249786 Another example of an aid is a battery-powered microchip memory with the following consumption For example, reading the last dose of the dose and/or reminding when to take the next dose of liquid crystal reader or voice alert signal. The kit of the present invention may include one or more other medicines in addition to the compounds of the present invention. Active compound. For example, 'other compounds are second antibacterial agents. Other compounds may be administered in the same dosage form as the compounds of the invention or in different dosage forms. Similarly, 'other compounds may be administered simultaneously or simultaneously with the compounds of the invention. The composition of the inventive compound may also be used in combination with other known antibacterial agents having a similar antimicrobial range to (1) enhance the treatment of severe Gram-negative infections covered by the antimicrobial range of the compound, or (2) increase the suspicion of multiple Coverage in severe infections of organisms, where another agent having a different antimicrobial range may be required in addition to this compound Possible agents include aglycosides, penicillins, cephai0Sp0rin, fluoroquinolone, macroslides, glycopeptides, lipopeptides, and oxazolidinone. member. Treatment may involve administration of a composition having a compound of the present invention and a second antibacterial compound or administration of a compound of the present invention, followed by or prior to administration of a second antibacterial agent. The above examples are better understood by reference to the following examples, which are presented for purposes of illustration and not limitation. EXAMPLES Referring to the following examples, the compounds of the invention were characterized by high performance liquid chromatography (HPLC) using a Waters Millenium chromatography system (Milford, Mass) with a 2690 separation module. The analytical column was an Alltima C-18 reverse phase tube 159849.doc • 39- 201249786 column, 4.6 x 250 mm from Alltech (Deerfield, 111.). A gradient elution starting with 5% acetonitrile/95% water and progressing to ι〇〇% acetonitrile over a period of 40 minutes was used. All solvents contain 〇 · 1 ° /. Trifluoroacetic acid (TFA). Compounds are detected by ultraviolet (UV) absorption at 220 nm or 254 nm. In some cases, the purity is evaluated by thin layer chromatography (TLC) using a glass or plastic substrate (such as Baker-Flex 1 B2-F flexible sheet). TLC results are readily visually detectable under ultraviolet light or by the use of well-known vapors and various other staining techniques. Mass spectrometry was performed on one of two LCMS instruments: waters system (Alliance HT HPLC and Micromass ZQ mass spectrometer; column:

Eclipse XDB-C-18，2.1x50 mm ;溶劑系統：含5-95%(或 35-95%，或 65-95% 或 95-95%)乙腈之水（含 0.05% TFA);流速0.8 mL/m in ;分子量範圍500至1500;錐孔電壓20 V;管柱溫度 40°C )或 Hewlett Packard 系統（11〇〇系列 HPLC ;管柱：Eclipse XDB-C18，2.1x50 mm ;溶劑系統：含1-95% 乙腈之水（含0.05% TFA);流速0.4 mL/min;分子量範圍 150至850 ;錐孔電壓50 V ;管柱溫度30°C)。所有質量均報導為質子化母體離子之質量。 GCMS分析經Hewlet Packard儀（具有質量選擇性偵測器 5973之HP6890系列氣相層析儀；注射器體積：1 pL ;初始管柱溫度：50°C ;最終管柱溫度：250°C ;勻變時間：20 分鐘；氣體流速：1 mL/min ;管柱：5%苯基甲基石夕氧烧， #HP 190915-443型，尺寸：30.0 m><25 mx〇.25 m)進行。核磁共振（NMR)分析由 Varian 300 MHz NMR(Palo Alto, 159849.doc -40- a 201249786Eclipse XDB-C-18, 2.1x50 mm; solvent system: 5-95% (or 35-95%, or 65-95% or 95-95%) acetonitrile in water (0.05% TFA); flow rate 0.8 mL /m in ; molecular weight range 500 to 1500; cone voltage 20 V; column temperature 40 ° C) or Hewlett Packard system (11 〇〇 series HPLC; column: Eclipse XDB-C18, 2.1 x 50 mm; solvent system: containing 1-95% acetonitrile water (0.05% TFA); flow rate 0.4 mL/min; molecular weight range 150 to 850; cone voltage 50 V; column temperature 30 °C). All masses are reported as the mass of the protonated parent ion. GCMS analysis by Hewlet Packard instrument (HP6890 series gas chromatograph with mass selective detector 5973; syringe volume: 1 pL; initial column temperature: 50 ° C; final column temperature: 250 ° C; Time: 20 minutes; gas flow rate: 1 mL/min; column: 5% phenylmethyl aspartic acid, #HP 190915-443 type, size: 30.0 m><25 mx 〇.25 m). Nuclear magnetic resonance (NMR) analysis by Varian 300 MHz NMR (Palo Alto, 159849.doc -40-a 201249786

Calif.)進行。光譜參照物為TMS或該溶劑之已知化學位移。在高溫（例如75。〇下運作一些化合物樣品以促進樣品溶解度提高。一些本發明化合物之純度藉由元素分析（DesertCalif.). The spectral reference is TMS or the known chemical shift of the solvent. Run some compound samples at elevated temperatures (eg, 75 〇 to promote sample solubility). Some of the compounds of the invention are purified by elemental analysis (Desert)

Analytics, Tucson, Ariz.)評估〇經Laboratory Devices Mel-Temp裝置（Holliston，Mass.)測定溶點。使用 Flash 40 層析系統及 Kp-Si丨，60A(Bi〇tage，Analytics, Tucson, Ariz.) Evaluation 溶 The melting point was determined by the Laboratory Devices Mel-Temp device (Holliston, Mass.). Using Flash 40 chromatography system and Kp-Si丨, 60A (Bi〇tage,

Charlottesville，va.)或藉由使用矽膠（23〇至4〇〇目）填充材料之急驟管柱詹析’或藉由使用C-18逆相管柱之HPLC來進行製備型分離。用於Flash 40 Biotage系統及急驟管柱層析之典型溶劑為二氣甲烷、甲醇、乙酸乙酯、己烷、丙酮、經胺水溶液及三乙胺。用於逆相HPLC之典型溶劑為不同濃度之乙腈及水（含^。/。三氟乙酸）。 A.合成N-((S)-3-胺基4-(羥胺基）_3_甲基_][_側氧基丁 _2_ 基）-4-(((1,2·反）-2-(羥甲基）環丙基）丁 4,3-二炔基）苯甲醯胺（1-1)Charlottesville, va.) or preparative separation by HPLC using a silicone (23 Å to 4 )) filling material or by HPLC using a C-18 reverse phase column. Typical solvents for the Flash 40 Biotage system and flash column chromatography are di-methane, methanol, ethyl acetate, hexane, acetone, aqueous amines and triethylamine. Typical solvents for reverse phase HPLC are different concentrations of acetonitrile and water (containing trifluoroacetic acid). A. Synthesis of N-((S)-3-amino 4-(hydroxyamino)_3_methyl_][_sideoxybut-2-yl)-4-(((1,2·re))-2 -(hydroxymethyl)cyclopropyl)butyl 4,3-diynyl)benzamide (1-1)

2-乙炔基環丙炫f酸甲酯（2) 在Nz下將外消旋2-甲醯基環丙烷曱酸乙酯i(i〇 g，70.3 mmol)及貝斯特曼大平試劑（Bestmann Ohira reagent)(16.4 g，85 mmol)溶解於無水甲醇（loo爪1)中。接著緩慢逐份添 159849.doc -41 - 201249786 加碳酸卸（19.4 g，141 mmol)且攪拌溶液18小時。在20°C 下減壓移除溶劑，添加水（100 ml)且用二氣曱烷（2x200 ml) 萃取產物，經硫酸鈉乾燥，且緩慢濃縮，得到藉由NMR確認之外消旋2-乙炔基環丙烷甲酸甲酯（4.16 g)。 (2·乙炔基環丙基）甲醇（3) 接著在氮氣下將棚氫化裡（175 mg，8.06 mmol)緩慢添加至含外消旋2-乙炔基環丙烷曱酸甲酯（1 g，20.6 mmol)之無水THF(20 ml)中，且持續攪拌2小時。用幾滴乙酸淬滅反應混合物且移除溶劑。用乙酸乙酯（2x50 ml)萃取粗產物，經硫酸鈉乾燥且緩慢濃縮，得到呈黃色液體狀之外消旋（2-反-乙炔基環丙基）甲醇3(73 5 mg)且其以原樣用於下一步驟。2-Ethynylcyclopropane f-acid methyl ester (2) Racemic 2-methylmercaptocyclopropanoic acid ethyl i (i〇g, 70.3 mmol) and Bestmann Daping reagent (Bestmann Ohira) under Nz Reagent) (16.4 g, 85 mmol) was dissolved in anhydrous methanol (loos 1). Then slowly add 159849.doc -41 - 201249786 by carbonic acid (19.4 g, 141 mmol) and stir the solution for 18 hours. The solvent was removed under reduced pressure at 20 ° C, EtOAc (EtOAc)EtOAc. Methyl ethynylcyclopropanecarboxylate (4.16 g). (2. ethynylcyclopropyl)methanol (3) Next, the shed hydrogenation (175 mg, 8.06 mmol) was slowly added to the methyl group containing racemic 2-ethynylcyclopropanoate (1 g, 20.6) under nitrogen. Methyl acetate in THF (20 ml) was stirred for 2 h. The reaction mixture was quenched with a few drops of acetic acid and the solvent was removed. The crude product was extracted with EtOAc (EtOAc (EtOAc) (EtOAcjjjjjj Used as is for the next step.

第三丁氧羰基胺基）-2-(4-((2-(羥甲基）環丙基）丁-1,3-二炔基）苯甲醯胺基）-3-甲基丁酸甲酯（5) 在氮氣下將CuCl(42 mg，0.416 mmol)緩慢添加至授拌之外消旋（2-反-乙炔基環丙基）甲醇（400 mg，4.16 mmol)、2-(S)-(4-(溴乙炔基）苯曱醯胺基）-3-(第三丁氧羰基胺基）-3-甲基丁酸曱酯（1.9 g，4.16 mmol)於 THF(20 ml)、曱醇（10 ml)Third butoxycarbonylamino)-2-(4-((2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzimidamide)-3-methylbutyric acid Methyl ester (5) CuCl (42 mg, 0.416 mmol) was slowly added under nitrogen to racemic racemic (2-trans-ethynylcyclopropyl)methanol (400 mg, 4.16 mmol), 2-(S -(4-(Bromoethynyl)phenylhydrazino)-3-(t-butoxycarbonylamino)-3-methylbutanoate (1.9 g, 4.16 mmol) in THF (20 ml) , sterol (10 ml)

159849.doc -42- S 201249786 與丁胺（10 ml)之混合物中之溶液中，之後添加幾個羥胺鹽酸鹽晶體（30 mg)。持續攪拌4小時。減壓移除溶劑，添加水（100 ml)且用乙酸乙酯（2xl5〇 mi)萃取，經硫酸鈉乾燥，濃縮且經ISCO純化，得到432 mg (2-S)-3-(第三丁氧幾基胺基）-2-(4-((2-(羥曱基）環丙基）丁 a，3_二炔基）苯曱醯胺基）-3-曱基丁酸甲酯。 1-1 : 胺基-U(羥胺基）甲基-側氧基丁_2-基）· 4-(((U2反）-2-(羥甲基）環丙基）丁_13·二炔基）笨甲醢胺將（2-S)-3-(第三丁氧羰基胺基）_2_(4_((2_(羥曱基）環丙基）丁 -1，3-二炔基）苯曱醯胺基）_3_曱基丁酸曱酯5(〇 37〇 g，0.814 mmol)溶解於二氣甲烷（25 ml)中，用 TFA(2 ml)處理且攪拌20分鐘。減壓移除過量溶劑及TFA，得到脫除保護基之物質’再溶解於][PA(1〇 mL)中且用羥胺水溶液 (5〇%，2 mL，過量）處理且保存於冰箱中2天。減壓移除過量溶劑’且藉由逆相HPLC純化粗產物’得到42 mg呈三氟乙酸鹽形式之1_1。LC-MS (M+1) 370，化學式： c20h23n3o4，精確質量：369 17。iH NMR (DMS〇〇, TFA鹽：δ 0.9 (m，2H), 1.24 (s，3H)，1.29 (s，3H)，1.40(m, 1H)，3.24 (m，1H)，3.4 (m, 1H)，4.64 (d，1H)，4.69 (d，1H), 7.61 (d, 2H), 7.88 (d, 2H), 7.90 (br.d, 1NH), 8.65 (d, 1NH), 9.1 (br.S，OH)。 Β·合成N-((S)-3-胺基羥胺基）·3_甲基側氧基丁 _2_ 基）-4-(((lR，2R)-2-(羥甲基）環丙基^丁“，、二炔基）苯甲醯胺（1-2) 159849.doc 43· 201249786159849.doc -42- S In a solution of a mixture of 201249786 and butylamine (10 ml), several hydroxylamine hydrochloride crystals (30 mg) were added. Stirring was continued for 4 hours. The solvent was removed under reduced pressure. EtOAc (EtOAc) (EtOAcjjjjjjjj Methylaminoamino)-2-(4-((2-(hydroxy))cyclopropyl)buta,3-diynyl)phenylhydrazinyl)-3-mercaptobutyrate. 1-1 : Amino-U (hydroxylamino)methyl-o-oxybutan-2-yl)· 4-(((U2 trans)-2-(hydroxymethyl)cyclopropyl)butane _13·II Alkynyl)(2-S)-3-(t-butoxycarbonylamino)_2_(4_((2-(hydroxyphenyl)cyclopropyl)butane-1,3-diynyl) Phenylguanidino) _3_mercaptobutyrate 5 (〇37 〇g, 0.814 mmol) was dissolved in di-methane (25 ml), taken with TFA (2 ml) and stirred for 20 min. Excess solvent and TFA were removed under reduced pressure to give the deprotected material 're-dissolved in' [PA (1 mL) and treated with aqueous hydroxylamine (5%, 2 mL, excess) and stored in the refrigerator. day. The excess solvent was removed under reduced pressure and the crude product was purified by reverse phase HPLC to afford &lt LC-MS (M+1) 370, chemical formula: c20h23n3o4, exact mass: 369 17. iH NMR (DMS 〇〇, TFA salt: δ 0.9 (m, 2H), 1.24 (s, 3H), 1.29 (s, 3H), 1.40 (m, 1H), 3.24 (m, 1H), 3.4 (m, 1H), 4.64 (d, 1H), 4.69 (d, 1H), 7.61 (d, 2H), 7.88 (d, 2H), 7.90 (br.d, 1NH), 8.65 (d, 1NH), 9.1 (br .S, OH). 合成·Synthesis of N-((S)-3-aminohydroxylamino)·3_methyl-oxo-butan-2-yl)-4-(((lR,2R)-2-(((lR,2R)-2-) Hydroxymethyl)cyclopropyl^butyl ",,diynyl)benzamide (1-2) 159849.doc 43· 201249786

(E)-乙酸4-羥基丁-2·烯酯（2) 試劑 MW 當量 mol g，mL 化合物1 88.05 1 3 264 g NaH(60°/〇) 24.00 1 3 120.0 g CH3COCI 78.50 1 3 235.5 g THF 1.5 L 在-20°C 下向（Ε)-丁-2-烯-1，4-二醇 1(264 g，3.0 mol)於 1.5 L THF中之溶液中逐份添加氫化鈉（120 g，3.0 mol)。添加後，混合物在-20°C下持續攪拌30分鐘。接著逐滴添加乙醯氣（23 5.5 g，3 mol)，使混合物升溫至室溫且在室溫下再持續攪拌3小時。過濾混合物且用THF洗滌殘餘物。乾燥經合併之有機層且濃縮，得到粗物質2，藉由矽膠管柱 (PE:EA=5:1至2:1)純化，得到210 g呈無色油狀之2。產率：54%。4 NMR:CP-0005065-043 (CDC13, 400 ΜΗΖ) δ: 5.85 (m, 1H), 5.62 (m, 1H), 4.67 (t, J=6.2 Hz, 2H), 4.26 (t, J=6.0 Hz，2H)，2.10 (s，1H), 2.06 (s，3H)。 159849.doc -44- 201249786(E)-Hydroxy-4-hydroxybutan-2-enyl acetate (2) Reagent MW Equivalent mol g, mL Compound 1 88.05 1 3 264 g NaH (60°/〇) 24.00 1 3 120.0 g CH3COCI 78.50 1 3 235.5 g THF 1.5 L To a solution of (Ε)-but-2-ene-1,4-diol 1 (264 g, 3.0 mol) in 1.5 L of THF at -20 ° C, add sodium hydride (120 g, 3.0 mol). After the addition, the mixture was continuously stirred at -20 ° C for 30 minutes. Ethylene gas (23 5.5 g, 3 mol) was then added dropwise, and the mixture was allowed to warm to room temperature and stirring was continued for 3 hours at room temperature. The mixture was filtered and the residue was washed with THF. The combined organic layers were dried and concentrated to give EtOAc (EtOAc: EtOAc) Yield: 54%. 4 NMR: CP-0005065-043 (CDC13, 400 ΜΗΖ) δ: 5.85 (m, 1H), 5.62 (m, 1H), 4.67 (t, J=6.2 Hz, 2H), 4.26 (t, J=6.0 Hz , 2H), 2.10 (s, 1H), 2.06 (s, 3H). 159849.doc -44- 201249786

(E)-己酸4·侧氧基丁-2-烯酯（3) 試劑 MW 當量 mol g，mL 化合物2 130.14 1 1.5 195 g Mn〇2(活性） 86.94 10 15 1305 g DCM 3L 向二氧化猛（活性，1305 g，15 mol)於2.5 L二氣甲炫中之懸浮液中逐份添加（E)-乙酸4-羥基丁 -2-烯酯2(195 g)。混合物在室溫下持續攪拌48小時。過濾混合物且用二氯曱烷洗滌殘餘物。乾燥經合併之有機層且濃縮，得到粗物質 3，藉由矽膠管柱（ρε··ΕΑ=10··1至5··1)純化，得到130 g呈無色油狀之3。產率：64%» iHNMR:CP-0005065_044(CDCl3, 400 MHZ) δ: 10.01 (d, J=6.4 Hz, 1H), 6.52 (m, 1H), 6.10 (m，1H)，5.08 (m，2H)，2.10 (s，3H) 〇 (E)-乙酸4,4-二乙氧基丁-2-烯酯（4)(E)-hexanoic acid 4 · oxoxybut-2-enyl ester (3) Reagent MW equivalent mol g, mL Compound 2 130.14 1 1.5 195 g Mn〇2 (active) 86.94 10 15 1305 g DCM 3L Dioxide (E)- 4-hydroxybut-2-enyl acetate 2 (195 g) was added portionwise in a suspension of vigorous (active, 1305 g, 15 mol) in 2.5 L of dioxane. The mixture was stirred continuously for 48 hours at room temperature. The mixture was filtered and the residue was washed with dichloromethane. The combined organic layers were dried and concentrated to give a crude material (yield: 3). Yield: 64%» iHNMR: CP-0005065_044 (CDCl3, 400 MHZ) δ: 10.01 (d, J = 6.4 Hz, 1H), 6.52 (m, 1H), 6.10 (m, 1H), 5.08 (m, 2H) ), 2.10 (s, 3H) 〇 (E)-acetic acid 4,4-diethoxybut-2-enyl ester (4)

試劑 MW 當量 mol g，mL 化合物3 128.13 1 0.75 96 g 三乙氧曱烷 148.20 1.2 0.90 133.2 g NH4NO3 79.90 0.05 0.038 3.0 g EtOH 500 mL 向（E)-乙酸4-側氧基丁-2-烯酯3(96.0 g，〇·75 m〇1)及= 乙氧甲烧（133.2 g，0.9 mol)於500 ml乙醇中之溶液中添加硝酸銨（3.0 g，0.038 mol)，混合物在室溫下持續攪拌15小時。用800 ml EtOAc稀釋混合物且用飽和碳酸氫鈉洗滌。用EtOAc(300 mlx2)反萃取水層，乾燥經合併之有機層且濃縮得到140 g呈紅色油狀之粗物質4，其未經進一步純化 159849.doc • 45- 201249786 即用於下一步驟。 (4R，5R,e)-2-(3-乙逋氧基丙-I •烯基）-1，3-二氧雜環戏烷-4,5-二▼酸二異丙酯（5) 試劑 MW 當量 mol g ' mL 化合物4 202.25 1 0.3 60.6 g (2R，3R)-2,3-二羥基丁二酸二異丙酯 234.25 1.1 0.33 77.2 g PPTS 251.09 0.05 0.015 3.8 g 苯 500 ml 向（Ε)-乙酸4,4-二乙氧基丁-2-烯酯4(60.6 g 0.3 mol)及 (2R,3R)-2,3-二羥基丁二酸二異丙酯（77·2 g，0.33 mol)於 500 ml苯中之溶液中添加PPTS(3.8 g，15 mmol)，加熱混合物至90°C以蒸餾出乙醇，維持15小時。使混合物冷卻至室溫且真空濃縮。藉由矽膠用（PE:EA=50:1至30:1)純化，獲得38.5 g呈無色油狀之5。產率：37.3%。GCMS:CP-0005065-070-2 (85%.^) ° (4R，5R)-2-((lS，2R〉-2-(乙醯氧基 f 基）環丙基）-1，3-二氧藉環戊烷-4,5-二甲酸二異丙酯（6}Reagent MW Equivalent mol g, mL Compound 3 128.13 1 0.75 96 g Triethoxy decane 148.20 1.2 0.90 133.2 g NH4NO3 79.90 0.05 0.038 3.0 g EtOH 500 mL To (E)-acetic acid 4-sided oxybut-2-enyl ester 3 (96.0 g, 〇·75 m〇1) and = ethoxylated (133.2 g, 0.9 mol) in 500 ml of ethanol, ammonium nitrate (3.0 g, 0.038 mol) was added and the mixture was continued at room temperature. Stir for 15 hours. The mixture was diluted with EtOAc (EtOAc) (EtOAc) The aqueous layer was re-extracted with EtOAc (EtOAc (EtOAc) (EtOAc). (4R,5R,e)-2-(3-Ethyloxypropan-I • alkenyl)-1,3-dioxetane-4,5-di-dicarboxylic acid diisopropyl ester (5) Reagent MW equivalent mol g ' mL Compound 4 202.25 1 0.3 60.6 g (2R,3R)-2,3-Dihydroxysuccinic acid diisopropyl 234.25 1.1 0.33 77.2 g PPTS 251.09 0.05 0.015 3.8 g Benzene 500 ml to (Ε - 4,4-diethoxybut-2-enyl acetate 4 (60.6 g 0.3 mol) and (2R,3R)-2,3-dihydroxysuccinate diisopropyl ester (77·2 g, 0.33 mol) PPTS (3.8 g, 15 mmol) was added to a solution of 500 ml of benzene, and the mixture was heated to 90 ° C to distill off ethanol for 15 hours. The mixture was allowed to cool to room temperature and concentrated in vacuo. Purification by silica gel (PE: EA = 50:1 to 30:1) gave 38.5 g of 5 as a colorless oil. Yield: 37.3%. GCMS: CP-0005065-070-2 (85%.^) ° (4R,5R)-2-((lS,2R>-2-(ethyloxyl)cyclopropyl)-1,3- Dioxycyclopentane-4,5-dicarboxylic acid diisopropyl ester (6}

試劑 MW 當量 mol g , mL 化合物5 344.36 1 0.1 34.4 g ZnEt2(l M己烧溶液） 5 0.5 500 ml CH2I2 267.84 10 1 267.8 g 己烷 1.5 L 在氬氣下在-20°C下向（4R,5R，E)-2-(3-乙醯氧基丙-1-烯基）-1，3-二氧雜環戊烷-4,5-二曱酸二異丙酯5(34.4 g，0.1 mol)於己烷（1.5 L)中之溶液中逐份添加二乙基鋅（1 Μ己烷溶液，500 mL)。添加後，低於-20°C下在強烈攪拌下逐滴 159849.doc • 46 - 201249786 添加二碘乙烷。使混合物升溫至室溫且再持續攪拌8小時。藉由800 ml冷氣化銨水溶液淬滅反應混合物，接著用乙趟（800 ml><5)萃取。用硫代硫酸鈉水溶液、水鹽水洗滌經合併之有機層，接著乾燥且濃縮，得到粗物質6，藉由石夕膠管柱（PE:EA=30:1至1〇:1)純化得到16 g呈無色油狀之6。產率：44.7%。*H NMR:P05 (CDC13, 400 ΜΗΖ) δ: 5.13 (m, 2Η), 4.95 (d, J=5.6 Hz, 1H), 4.67 (d, J=3.6 Hz, 1H), 4.57 (d, J=4.0 Hz, 1H), 4.07 (m, 1H), 3.88 (m, 1H), 2.06 (s5 3H), 1.38 (s, 12H), 1.23 (m, 1H)S 0.83 (m, 1H), 0.66 (m，1H)。乙酸（(lR，2R)-2- f M基環丙基）f酯（7)Reagent MW equivalent mol g , mL Compound 5 344.36 1 0.1 34.4 g ZnEt2 (l M hexane solution) 5 0.5 500 ml CH2I2 267.84 10 1 267.8 g Hexane 1.5 L Under argon at -20 ° C (4R, 5R,E)-2-(3-Ethyloxyprop-1-enyl)-1,3-dioxolane-4,5-didecanoic acid diisopropyl ester 5 (34.4 g, 0.1 Methyl) Diethylzinc (1 hexane solution, 500 mL) was added portionwise to a solution of hexane (1.5 L). After the addition, drop below -20 ° C with vigorous stirring 159849.doc • 46 - 201249786 Add diiodoethane. The mixture was allowed to warm to room temperature and stirring was continued for an additional 8 hours. The reaction mixture was quenched by 800 ml of cold aqueous ammonium sulfate, and then extracted with ethyl acetate (800 ml < 5 <5>). The combined organic layers were washed with aqueous sodium thiosulfate solution and brine, then dried and concentrated to afford crude material 6 which was purified by EtOAc (EtOAc: EA=30:1 to 1:1:1) 6 in the form of a colorless oil. Yield: 44.7%. *H NMR: P05 (CDC13, 400 ΜΗΖ) δ: 5.13 (m, 2Η), 4.95 (d, J=5.6 Hz, 1H), 4.67 (d, J=3.6 Hz, 1H), 4.57 (d, J= 4.0 Hz, 1H), 4.07 (m, 1H), 3.88 (m, 1H), 2.06 (s5 3H), 1.38 (s, 12H), 1.23 (m, 1H)S 0.83 (m, 1H), 0.66 (m , 1H). Acetic acid ((lR, 2R)-2- f M-cyclopropyl)f ester (7)

試劑 MW 當量 mmol g j mL 化合物6 AcOHf80%l 358.38 1 40 14.3 g 140 mL 將（4R，5R)-2-((lS，2R)-2-(乙醯氧基曱基）環丙基）] 氧雜壤戊烧-4,5-·一曱酸二異丙醋6(14.3 g，40 mmol)於140 ml 80°/〇乙酸中之混合物加熱至8〇它’且在此溫度下持續授拌2小時。當TLC顯示剩餘少量6時，將混合物逐滴添加至 300 ml飽和碳酸氫鈉中’接著用二氣甲烷（2〇〇 mix3)萃取。用水、鹽水洗滌經合併之有機層，乾燥且濃縮，得到粗物質7 ’藉由矽膠管柱（pe:EA=10:1至5:1)純化得到3.5 g 呈無色油狀之 7。產率：62%。4 NMR:CP-0005065-072 (CDC13, 400 MHZ) δ: 9.15 (s，1H)，4.11 (m，1H)，3.91 (m， 1H)，2.08 (s，3H)，2.10(S，3H)，1.88 (m，2H)，1.39 (m, iH)， 159849.doc •47- 201249786 1.12 (m，1H)。乙酸（(lR，2R)-2-(2,2-二溴乙烯基）環丙基）甲酯（8)Reagent MW equivalent mmol gj mL Compound 6 AcOHf 80% l 358.38 1 40 14.3 g 140 mL Will (4R,5R)-2-((lS,2R)-2-(ethyloxycarbonyl)cyclopropyl)oxy Mixture of miscellaneous pentylene-4,5--monodecanoic acid diisopropyl vinegar 6 (14.3 g, 40 mmol) in 140 ml of 80 ° / hydrazine acetic acid to 8 〇 it and continue to mix at this temperature 2 hours. When TLC showed a small amount of 6 remaining, the mixture was added dropwise to 300 ml of saturated sodium hydrogencarbonate' and then extracted with dioxane (2 〇〇 mix 3). The combined organic layers were washed with water and brine, dried and evaporated tolulululululululululululululululululululululu Yield: 62%. 4 NMR: CP-0005065-072 (CDC13, 400 MHZ) δ: 9.15 (s, 1H), 4.11 (m, 1H), 3.91 (m, 1H), 2.08 (s, 3H), 2.10 (S, 3H) , 1.88 (m, 2H), 1.39 (m, iH), 159849.doc • 47- 201249786 1.12 (m, 1H). Acetic acid ((lR, 2R)-2-(2,2-dibromovinyl)cyclopropyl)methyl ester (8)

試劑 MW 當量 mmol g，mL 化合物7 142.15 1 21 3.00 g CBr4 331.63 2 42 13.9g PPh3 262.29 4 84 22.0 g DCM 120 mL 在-20°C下在氬氣下向四溴化碳（13.9 g，42 mmol)於30 mL二氣曱烷中之溶液中逐滴添加三苯膦（22.0 g，84 mmol) 於50 mL二氯甲烷中之溶液。混合物在此溫度下持續攪拌半小時，接著冷卻至-78°C。逐滴添加乙酸（（lR,2R)-2-甲醯基環丙基）甲醋7(3.00 g，21 mmol)於40 ml二氣曱烧中之溶液，再維持於該溫度下半小時。經30分鐘使混合物升溫至室溫。移除溶劑且藉由矽膠管柱（PE:EA=10(hl至50:1)純化，得到4.3 g呈無色油狀之8。產率：69%。4 NMR:CP-0005065-075(CDCl3, 400 MHZ) δ: 5.85 (d, J=8.8Hz, 1H), 3.98 (m, 2H), 2.09 (m, 3H), 1.61 (m, 1H), 1.35 (m,lH), 0.88 (m，2H)。 4-(((lR，2R)-2-(乙醯氡基甲基）環丙基）丁-1，3·二炔基）苯甲酸甲酯（10)Reagent MW equivalent mmol g, mL Compound 7 142.15 1 21 3.00 g CBr4 331.63 2 42 13.9g PPh3 262.29 4 84 22.0 g DCM 120 mL At -20 ° C under argon to carbon tetrabromide (13.9 g, 42 mmol A solution of triphenylphosphine (22.0 g, 84 mmol) in 50 mL of dichloromethane was added dropwise to a solution of 30 mL of dioxane. The mixture was stirred at this temperature for a half hour and then cooled to -78 °C. A solution of acetic acid ((lR, 2R)-2-methylindenylcyclopropyl)methine 7 (3.00 g, 21 mmol) in 40 ml of dioxane was added dropwise, and maintained at this temperature for half an hour. The mixture was allowed to warm to room temperature over 30 minutes. The solvent was removed and purified by a silica gel column (PE: EA = 10 (hl to 50:1) to afford 4.3 g as colorless oil. Yield: 69%. 4 NMR: CP-0005065-075 (CDCl3) , 400 MHZ) δ: 5.85 (d, J=8.8Hz, 1H), 3.98 (m, 2H), 2.09 (m, 3H), 1.61 (m, 1H), 1.35 (m, lH), 0.88 (m, 2H) 4-(((lR,2R)-2-(ethyl)methyl)cyclopropyl)butane-1,3.diynyl)benzoic acid methyl ester (10)

試劑 MW 當量 mmol g，mL 化合物8 297.97 1 14.5 4.3 g 化合物9 160.17 1.1 16.0 2.56 g Pd2dba2 575.22 0.01 0.15 86.3 mg TMPP 352.24 0.04 0.58 204.2 mg NEt3 101.19 3 43.5 4.35 g DMF 100 mL 159849.doc •48· 201249786 在氬氣下乙酸（（lR，2R)-2-(2,2-二溴乙烯基）環丙基）曱酯 8(4.3 g，14.5 mmol)、Pd2<lba2(86.3 mg，0.15 mmol)、三 (4-甲基苯基）膦（204 mg，0.58 mmol)、三乙胺（4·35 g， 43.5 mmol)於DMF 100 mL中之溶液用4_乙炔基苯曱酸曱酯 9(2.64 g，11 mmol)處理。混合物在室溫下持續授拌5小時。當TLC顯示剩餘少量化合物8時’用Et0Ac(300 mL)稀釋反應物且用水（3 X 100 mL)洗滌’乾燥有機層且濃縮’得到粗物質10，藉由矽膠管柱層析（PE:EA=50:1至30:1)純化，得到2.0 g呈黃色固體狀之10。產率：46.5°/〇， LCMS:CP-0005065-085-2 (ESI) m/z=297 (M+1)純度： 4-(((lR，2R>-2-(羥甲基> 環丙基）丁-1，3·二炔基）苯甲酸（11) 92.4% (214 nm) °Reagent MW equivalent mmol g, mL Compound 8 297.97 1 14.5 4.3 g Compound 9 160.17 1.1 16.0 2.56 g Pd2dba2 575.22 0.01 0.15 86.3 mg TMPP 352.24 0.04 0.58 204.2 mg NEt3 101.19 3 43.5 4.35 g DMF 100 mL 159849.doc •48· 201249786 Acetic acid ((lR,2R)-2-(2,2-dibromovinyl)cyclopropyl)decyl 8 (4.3 g, 14.5 mmol), Pd2 <lba2 (86.3 mg, 0.15 mmol), three under argon a solution of (4-methylphenyl)phosphine (204 mg, 0.58 mmol), triethylamine (4·35 g, 43.5 mmol) in DMF 100 mL with ethyl 4-ethynylbenzoate 9 (2.64 g) , 11 mmol) treatment. The mixture was continuously mixed for 5 hours at room temperature. When TLC showed a small amount of compound 8 remaining, the reaction was diluted with Et0Ac (300 mL) and washed with water (3×100 mL) and then dried and concentrated to afford crude material 10 by column chromatography (PE: EA) Purification from =50:1 to 30:1) gave 2.0 g as a yellow solid. Yield: 46.5°/〇, LCMS: CP-0005065-085-2 (ESI) m/z=297 (M+1) Purity: 4-(((lR,2R>-2-(hydroxymethyl)> Cyclopropyl) butyl-1,3·diynyl)benzoic acid (11) 92.4% (214 nm) °

試劑 MW 當量 mmol g 5 mL 化合物10 296.32 1 6.5 1.92 g NaOH THF 40.10 10 65 2.60 g 40 mL 將4-(((lR,2R)_2-(乙醯氧基甲基）環丙基）T_i，3_二炔基）苯甲酸甲酯10(1.92呂，6.5 111111〇1)溶解於丁11?(4〇1111^)中，接著添加氫氧化鈉（2.60 g ’ 65 mmol)於10 "^水中之溶液。混合物在室溫下持續攪拌8小時。當LCMS顯示剩餘少量化合物10時，減壓移除溶劑’用水（5〇 mL)稀釋殘餘物’將pH值調節至4.0 ’用乙酸乙醋（4x50 mL)萃取，乾燥有機層且濃縮，得到1.4 g呈黃色固體狀之粗物質11，其未經進一步純化即用於下一步驟。LCMS:CP-0005065-088-3 (ESI) m/z=241 (M+1)純度：89°/〇 (214 nm)。產率：89〇/〇。 159849.doc -49 - 201249786 (S)-2-(4-(((lR，2R)-2-(乙醯氧基 fS)環丙基）丁-1，3-二诀基）苯甲醯胺基(第三丁氧羰基胺基hk甲基丁酸甲輯 (12)Reagent MW equivalent mmol g 5 mL Compound 10 296.32 1 6.5 1.92 g NaOH THF 40.10 10 65 2.60 g 40 mL 4-(((lR,2R)_2-(ethyloxymethyl)cyclopropyl)T_i,3 _Diynyl) methyl benzoate 10 (1.92 LV, 6.5 111111 〇 1) was dissolved in butyl 11? (4〇1111^), followed by sodium hydroxide (2.60 g '65 mmol) in 10 " Solution. The mixture was kept stirring at room temperature for 8 hours. When the LCMS showed a small amount of compound 10 remaining, the solvent was removed under reduced pressure, and the residue was diluted with water (5 〇mL). The pH was adjusted to 4.0 ′, extracted with ethyl acetate (4×50 mL), the organic layer was dried and concentrated to give 1.4 The crude material 11 was obtained as a yellow solid, which was used in the next step without further purification. LCMS: CP-0005065-088-3 (ESI) m/z = 241 (M +1). Yield: 89 〇 / 〇. 159849.doc -49 - 201249786 (S)-2-(4-((lR,2R)-2-(ethyloxylS)cyclopropyl)butan-1,3-didecyl)benzamide Amino (third butoxycarbonylamino) hk methylbutyric acid A series (12)

試劑 MW 當量 mmol g，mL 化合物11 240.25 1 5.0 1.20 g Boc-二-Me-DAP 246.30 1.2 6.0 1.48 g HATU 390.12 1.2 6.0 2.34 g DIPEA 129.24 4 20 3.58 g DMF 50 mL 4-(((lR，2R)-2-(羥甲基）環丙基）丁 -1，3-二炔基）苯甲酸 11(1.20 g，5.0 mmol)、HATU(2.34 g，6 mmol)於 50 mL DMF中之溶液中用（S)-2-胺基-3-(第三丁氧羰基胺基）-3-甲基丁酸曱酯（1.48 g，6.0 mmol)及 DIPEA(3.58 g，20 mmol) 處理。混合物在室溫下持續攪拌5小時。當LCMS顯示剩餘少量化合物11時，用EtOAc(100 mL)稀釋反應物，用5%氯化鋰（3x50 mL)洗滌，乾燥有機層且濃縮，得到呈黃色油狀之12。藉由矽膠管柱層析（PE:EA=2:1)純化，得到2.0 g 呈無色油狀之 12，產率：70%，LCMS:CP-0005065-091-3 (ESI) m/z=469 (M+1)，純度：95% (214 nm)。胺基-2-(4-(((lR，2R)-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺基）-3-甲基丁酸甲酯鹽酸鹽（U)Reagent MW equivalent mmol g, mL Compound 11 240.25 1 5.0 1.20 g Boc-di-Me-DAP 246.30 1.2 6.0 1.48 g HATU 390.12 1.2 6.0 2.34 g DIPEA 129.24 4 20 3.58 g DMF 50 mL 4-(((lR,2R) 2-(Hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzoic acid 11 (1.20 g, 5.0 mmol), HATU (2.34 g, 6 mmol) in 50 mL of DMF Treatment with (S)-2-amino-3-(t-butoxycarbonylamino)-3-methylbutanoate (1.48 g, 6.0 mmol) and DIPEA (3.58 g, 20 mmol). The mixture was kept stirring at room temperature for 5 hours. The residue was diluted with EtOAc (EtOAc) (EtOAc)EtOAc. Purification by hydrazine gel column chromatography (PE: EA = 2:1) afforded 2.0 g as a colorless oil. Yield: 70%, LCMS: CP-0005065-091-3 (ESI) m/z = 469 (M+1), purity: 95% (214 nm). Amino-2-(4-(((lR,2R)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzimidamide)-3-methylbutyric acid Methyl ester hydrochloride (U)

試劑 MW 當量 mmol g，mL 化合物12 468.23 1 4.0 1.87 g HCl(g) 36.5 CH3OH 50 mL 159849.doc -50- 201249786 將（8)-2-(4-(((111，211)-2-(乙酿氧基曱基）環丙基）丁_1，3_二炔基）苯甲醯胺基）-3-(第三丁氧羰基胺基）-3-曱基丁酸曱酯 12(1.87 g，4_0 mmol)溶解於曱醇（50 mL)中，用乾燥 HClg 處理10分鐘。當LCMS顯示剩餘少量化合物12時，停止HC1 流。減壓移除溶劑，且得到1.45 g呈黃色固體狀之13。產率：91%，LCMS: CP-0005065-096-4-LCMSA019 (ESI) m/z= 369 (M+l) ’ 純度：98%(214nm)。1HNMR:CP-0005065- 096-4(DMSO-400 MHz) δ:9.04 (d, J=6.8 Hz, 1H), 8.36 (s, 3H), 7.98 (d, J=6.4 Hz, 2H), 7.64 (d, J=6.8 Hz, 2H), 4.89 (d, J=6.8 Hz, 1H), 3.73 (s, 3H), 3.44 (m, 1H), 3.22 (m, 1H)，1.48 (m, 2H)，1.40 (s, 6H)，0.94 (m, 2H)。Reagent MW equivalent mmol g, mL Compound 12 468.23 1 4.0 1.87 g HCl (g) 36.5 CH3OH 50 mL 159849.doc -50- 201249786 (8)-2-(4-((111,211)-2-( Ethyloxycarbonyl)cyclopropyl)butyr-7,3-diynyl)benzamideamino)-3-(t-butoxycarbonylamino)-3-mercaptobutyrate decyl 12 1.87 g, 4_0 mmol) was dissolved in decyl alcohol (50 mL) and treated with dry HClg for 10 min. The HC1 stream was stopped when LCMS showed a small amount of compound 12 remaining. The solvent was removed under reduced pressure and 1.45 g was obtained as a yellow solid. Yield: 91%, LCMS: CP-0005065-096-4-LCMSA 019 (ESI) m/z = 369 (M+l) s purity: 98% (214nm). 1H NMR: CP-0005065- 096-4 (DMSO-400 MHz) δ: 9.04 (d, J = 6.8 Hz, 1H), 8.36 (s, 3H), 7.98 (d, J = 6.4 Hz, 2H), 7.64 ( d, J = 6.8 Hz, 2H), 4.89 (d, J = 6.8 Hz, 1H), 3.73 (s, 3H), 3.44 (m, 1H), 3.22 (m, 1H), 1.48 (m, 2H), 1.40 (s, 6H), 0.94 (m, 2H).

,nh3 (S)-3-胺基-2-(4-(((lR，2R)-2-(羥曱基）環丙基）丁·1>3-二快基）苯甲醯胺基）-3-甲基丁酸甲g旨鹽酸鹽13(1.25 g，3.1 mmol)用異丙醇（1〇 mL)及50%經胺水溶液（4.1 mL，61.7 mmol ’ 20當量）處理直至LCMS顯示處理顯著完成為止。藉由逆相HPLC(梯度為含0至30%乙腈之水（各自含有 0· 1 % TFA) ’歷經120分鐘）純化粗物質，且彙集所需溶離份且凍乾，得到呈三氟乙酸鹽形式之N-((s)-3-胺基-1-(羥胺基）-3-曱基-1-側氧基丁-2-基）-4-(((lR，2R)-2-(羥甲基）環 159849.doc -51 - 201249786 丙基）丁-1，3-二炔基）苯甲醯胺（白色固體，647 mg，1.3 mmol，43%)。質譜資料：預期值（M+1) : 370.4，觀測值：370.2。質子NMR (400 MHz，dmso-c?6): 11.20 (s，1H), 9.43 (br s, 1H), 8.55 (d, 1H, J=9.6 Hz), 8.00 (br s, 3H), 7.89 (dd, 2H, J=1.8, 6.6 Hz), 7.60 (dd, 2H, J=2.0, 6.8 Hz), 8.66 (d, 1H, J=9.2 Hz), 3.39 (dd, 1H, J=4.8, 11.6 Hz), 3.22 (dd, 1H, J=5.8, 11.4 Hz), 1.39-1.46 (m, 2H), 1.30 (s, 3H), 1.25 (s，3H), 0.84-0.93 (m，2H)。 C.合成N-((S)-3-胺基-1-(羥胺基）-3_甲基-1-側氧基丁-2-基）-4-(((lS，2S)-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺 (1-3) _乙@1^—> Mn〇a . 二乙氧基·甲 $, nh3 (S)-3-amino-2-(4-((lR,2R)-2-(hydroxyindenyl)cyclopropyl)butan-1>3-diyl)benzamide -3-methylbutyric acid methylglycol hydrochloride 13 (1.25 g, 3.1 mmol) was treated with isopropanol (1 mL) and 50% aqueous amine (4.1 mL, 61.7 mmol < 20 eq) until LCMS The display processing is significantly completed. The crude material was purified by reverse phase HPLC (gradient for water containing 0 to 30% acetonitrile (each containing 0.1% TFA) <RTI ID=0.0> Form of N-((s)-3-amino-1-(hydroxyamino)-3-indolyl-1-yloxybutan-2-yl)-4-(((lR,2R)-2- (Hydroxymethyl) ring 159849.doc -51 - 201249786 propyl)butyl-1,3-diynyl)benzamide (white solid, 647 mg, 1.3 mmol, 43%). Mass spectral data: expected (M+1): 370.4, observed: 370.2. Proton NMR (400 MHz, dmso-c?6): 11.20 (s, 1H), 9.43 (br s, 1H), 8.55 (d, 1H, J = 9.6 Hz), 8.00 (br s, 3H), 7.89 ( Dd, 2H, J=1.8, 6.6 Hz), 7.60 (dd, 2H, J=2.0, 6.8 Hz), 8.66 (d, 1H, J=9.2 Hz), 3.39 (dd, 1H, J=4.8, 11.6 Hz ), 3.22 (dd, 1H, J=5.8, 11.4 Hz), 1.39-1.46 (m, 2H), 1.30 (s, 3H), 1.25 (s, 3H), 0.84-0.93 (m, 2H). C. Synthesis of N-((S)-3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)-4-(((lS,2S)-2) -(Hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (1-3) _B @1^—> Mn〇a . Diethoxy·A $

NflH/THF ’ 〇 w ’ 〇 w NH^NOjiEtOH 0 丫 *1 2 3 4 0、^ (-)-(S，S)-酒石酸丨 PrO^C iPr〇2〇 =異丙& - 一厂？ CH,l,.EUn irf〇c. Λ9、 70%气 Λ _ ^ PPT& 苯.90〇C,7 小時 iPr〇2C”.（〇人〆~£,¾ 2 f S 6 O 0 7 0NflH/THF ’ 〇 w 〇 NH w NH^NOjiEtOH 0 丫 *1 2 3 4 0,^ (-)-(S,S)-tartaric acid 丨 PrO^C iPr〇2〇 =isopropyl & - One plant? CH,l,.EUn irf〇c. Λ9, 70% gas Λ _ ^ PPT& benzene.90〇C, 7 hours iPr〇2C”.(〇人〆~£,3⁄4 2 f S 6 O 0 7 0

UOH PPh^CBrt 人 r BrT"O~^_ …UOH PPh^CBrt people r BrT"O~^_ ...

Ο e Pd(PPhi)aCI2,Cul, DIPEA P 二-Me-DAP \H HATU, OIPEA* C>*" =Ο e Pd(PPhi)aCI2,Cul, DIPEA P II-Me-DAP \H HATU, OIPEA* C>*" =

NHBoc HC1NHBoc HC1

MeOH NHpHCI 'COjMe 次、 (4S，SS，E)-2-(3•乙蘸氧基丙-1-烯基）-1，3-二氧雜環戊烷-4,5-二甲酸二異丙酯（S) 試劑 MW 當量 mol g，mL 化合物4 202.25 1 0.37 75 g (-)-(S，S)-酒石酸二異丙酯 234.25 1 0.37 86.8 g PPTS 251.09 0.05 0.0185 4.66 g 笨 800 ml 159849.doc -52· 201249786 向（E)-乙酸4,4·二乙氧基丁烯酯4(75 g，0.37 mol)及 (_)-(S，S)-酒石酸一異丙醋（86.8 g，0.3 mol)於 800 ml苯中之溶液中添加PPTS(4.66 g，0.0185 mol)，加熱混合物至 9 C以蒸餾出乙醇，維持15小時。使混合物冷卻至室溫且真空濃縮。蒸餾純化得到呈無色油狀之化合物5(5〇〇 g， 39%) 〇 !H NMR: (CDC13j 400 MHz) 5:6.04-6.01 (m, 1H), 5.81-5.80 (m, 1H), 5.57 (d, J=8 Hz, 1H), 5.07-5.01 (m, 2H), 4.65 (d, J=4 Hz, 1H), 4.57 (d, J=4 Hz, 1H), 4.54-4.53 (m, 2H)，1·99 (s，3H), 1.23-1.19 (m, 12H)。 (4S，5S)-2-((lR，2S)-2·(乙醯氧基甲基）環丙基）-1>3•二氧雜環戊烷-4,5-二甲酸二異丙酯（6>MeOH NHpHCI 'COjMe, (4S,SS,E)-2-(3•Ethoxypropan-1-enyl)-1,3-dioxol-4,5-dicarboxylic acid diiso Propyl ester (S) reagent MW equivalent mol g, mL Compound 4 202.25 1 0.37 75 g (-)-(S,S)-diisopropyl tartrate 234.25 1 0.37 86.8 g PPTS 251.09 0.05 0.0185 4.66 g Stupid 800 ml 159849. Doc -52· 201249786 to (E)-acetic acid 4,4·diethoxybutene 4 (75 g, 0.37 mol) and (_)-(S,S)-tartaric acid-isopropyl vinegar (86.8 g, 0.3 mol) PPTS (4.66 g, 0.0185 mol) was added to a solution of 800 ml of benzene, and the mixture was heated to 9 C to distill off ethanol for 15 hours. The mixture was allowed to cool to room temperature and concentrated in vacuo. Purification by distillation afforded compound 5 (5 〇〇g, 39%) as a colorless oil. NMR: (CDC13j 400 MHz) 5:6.04-6.01 (m, 1H), 5.81-5.80 (m, 1H), 5.57 (d, J=8 Hz, 1H), 5.07-5.01 (m, 2H), 4.65 (d, J=4 Hz, 1H), 4.57 (d, J=4 Hz, 1H), 4.54-4.53 (m, 2H), 1·99 (s, 3H), 1.23-1.19 (m, 12H). (4S,5S)-2-((lR,2S)-2·(ethyloxymethyl)cyclopropyl)-1>3•dioxalan-4,5-dicarboxylic acid diisopropyl Ester (6>

試劑 MW 當量 mol g，mL 化合物5 344.36 1 0.12 40 g ZnEtyi Μ己烷溶液） 10 1.20 1.16L ch2i2 267.84 20 2.40 623 g 己烷 1.0 L 在氬氣下在-20°C下向（4S，5S，E)-2-(3-乙醯氧基丙-1-烯基）-1，3-二氧雜環戊烷_4,5_二甲酸二異丙酯5(4〇 g，〇12 mol)於己烷（1 .〇 l)中之溶液中逐份添加二乙基辞（丨μ己烷溶液’ 1.16 L)。添加後’低於_2〇ι下在強烈攪拌下逐滴添加二碘甲烷（623 g，2.40 mol)。使混合物升溫至室温且再持續攪拌8小時。藉由8〇〇 ml冷氣化銨水溶液淬滅反應混合物，接著用***（800 mlx5)萃取。用硫代硫酸鈉水溶液、水、鹽水洗滌經合併之有機層，接著乾燥且濃縮，得到粗化合物6，藉由矽膠管柱用含乙酸乙酯之石油醚（3%至 159849.doc •53· 201249786 10% v/v)純化得到呈無色油狀之化合物6(20.0 g，50°/〇)。 'Η NMR: (CDC13, 400 MHz) δ: 5.15-5.08 (m, 2H), 4.95 (d, */=5.6 Hz, 1H), 4.67 (d, /=3.6 Ηζ,ΙΗ), 4.57 (d, 7=4.0 Ηζ,ΙΗ), 4.07-4.04 (m,lH), 3.91-3.86 2.06(s,3H), 1.40-1.37 (m,lH), 1.31-1.28 (m,12H), 1.24-1.22 (m,lH), 0.85-0.82 (m，lH), 0.68-0.63 (m，lH)。己酸（(lS，2S)-2·甲醯基環丙基）甲酯（7)Reagent MW equivalent mol g, mL Compound 5 344.36 1 0.12 40 g ZnEtyi Μ hexane solution) 10 1.20 1.16L ch2i2 267.84 20 2.40 623 g Hexane 1.0 L Under argon at -20 ° C (4S, 5S, E)-2-(3-Ethyloxyprop-1-enyl)-1,3-dioxolane-4,5-dicarboxylic acid diisopropyl ester 5(4〇g, 〇12 mol To a solution of hexane (1. 〇l), diethyl ether (丨μhexane solution ' 1.16 L) was added portionwise. After the addition, diiodomethane (623 g, 2.40 mol) was added dropwise with vigorous stirring under _2 〇. The mixture was allowed to warm to room temperature and stirring was continued for another 8 hours. The reaction mixture was quenched with 8 mL of cold cold aqueous ammonium chloride and then extracted with diethyl ether (800 ml. The combined organic layers were washed with aqueous sodium thiosulphate, water, brine, and then dried and concentrated to give crude compound 6 EtOAc EtOAc EtOAc The compound 6 (20.0 g, 50 ° / 〇) was obtained as a colorless oil. 'Η NMR: (CDC13, 400 MHz) δ: 5.15-5.08 (m, 2H), 4.95 (d, */=5.6 Hz, 1H), 4.67 (d, /=3.6 Ηζ, ΙΗ), 4.57 (d, 7=4.0 Ηζ,ΙΗ), 4.07-4.04 (m,lH), 3.91-3.86 2.06(s,3H), 1.40-1.37 (m,lH), 1.31-1.28 (m,12H), 1.24-1.22 (m , lH), 0.85-0.82 (m, lH), 0.68-0.63 (m, lH). Caproic acid ((lS, 2S)-2.-mercaptocyclopropyl)methyl ester (7)

試劑 MW 當量 mmol g * mL 化合物6 AcOH(80%) 358.38 1 55.80 20.0 g 200 mL 加熱（4S，5S)-2-((lR，2S)-2-(乙醯氧基曱基）環丙基）4,3-二氧雜環戊烷-4,5-二曱酸二異丙酯6(2〇〇 g，55.8 mmol)於 200 ml 80°/。乙酸中之混合物至8〇°c，且在此溫度下持續攪拌2小時。當TLC顯示剩餘少量6時，用水（15〇 mL)稀釋混合物’用乙酸乙酯（2〇0 mlx3)萃取。用飽和碳酸氫鈉、水、鹽水洗滌經合併之有機層，接著乾燥且濃縮，得到粗化合物7，藉由矽膠管柱用含乙酸乙酯之石油醚（8%至2〇% v/v)純化得到呈無色油狀之化合物7(3 〇经，38%)。 NMR: (CDCI3, 400 MHz) δ: 9.15 (d, 7=4.8, 1H), 4.14-4.09 (m，1H)，3.95-3.90 (m，1H)，2.Ο8 (s, 3H)，1.92-1.88 (m， 2H)，1.38-1.36 (m，1H)，1.12-1.09 (m，m)。乙酸((lS，2S)-2-(2,2-:漠乙烯基)環丙基）甲賴⑻ 159849.doc 201249786Reagent MW equivalent mmol g * mL Compound 6 AcOH (80%) 358.38 1 55.80 20.0 g 200 mL Heated (4S,5S)-2-((lR,2S)-2-(ethyloxyfluorenyl)cyclopropyl 4,3-dioxolane-4,5-dioxalate diisopropyl 6 (2 〇〇g, 55.8 mmol) in 200 ml 80 ° /. The mixture in acetic acid was brought to 8 ° C and stirring was continued at this temperature for 2 hours. When TLC showed a small amount of 6 remaining, the mixture was diluted with water (15 〇 mL) and extracted with ethyl acetate (2 〇 0 ml x 3). The combined organic layers were washed with saturated sodium bicarbonate, water and brine, then dried and concentrated to afford crude compound 7 with ethyl ether petroleum ether (8% to 2% v/v) Purification afforded compound 7 (3 EtOAc, 38%). NMR: (CDCI3, 400 MHz) δ: 9.15 (d, 7=4.8, 1H), 4.14-4.09 (m, 1H), 3.95-3.90 (m, 1H), 2. Ο8 (s, 3H), 1.92 1.88 (m, 2H), 1.38-1.36 (m, 1H), 1.12-1.09 (m, m). Acetic acid ((lS, 2S)-2-(2,2-: desert vinyl) cyclopropyl) methyl lysine (8) 159849.doc 201249786

試劑 MW 當量 mmol g，mL 化合物7 142.15 1 21.00 3.00 g CBr4 331.63 2 42.00 13.90 g PPh3 262.29 4 84.00 22.00 g DCM 120 mL 在-20°C下在氬氣下向四溴化碳（13·9 g，42.0 mmol)於二氣甲烷（30 mL)中之溶液中逐滴添加三苯膦（22.0 g，84.0 mmol)於5 0 mL二氯甲烧中之溶液。混合物在此溫度下持續攪拌半小時，接著冷卻至_78°C。逐滴添加乙酸（（lS，2S)-2-曱醯基環丙基）曱酯7(3.0 g，21.0 mmol)於40 mL二氯甲烷中之溶液，再維持於該溫度下半小時。經30分鐘使混合物升溫至室溫。真空移除溶劑，且藉由矽膠管柱用石油醚純化殘餘物，得到呈無色油狀之化合物8(3.20 g，51%)。藉由GC-MS確認。 4-(((lS，2S)-2-(乙醯氧基甲基）環丙基）丁-1,3-二炔基）苯甲酸甲酯（1〇>Reagent MW equivalent mmol g, mL Compound 7 142.15 1 21.00 3.00 g CBr4 331.63 2 42.00 13.90 g PPh3 262.29 4 84.00 22.00 g DCM 120 mL At -20 ° C under argon to carbon tetrabromide (13·9 g, 42.0 mmol) A solution of triphenylphosphine (22.0 g, 84.0 mmol) in 50 mL of methylene chloride was added dropwise to a solution in di-methane (30 mL). The mixture was stirred at this temperature for half an hour and then cooled to -78 °C. A solution of acetic acid ((lS, 2S)-2-mercaptocyclopropyl) decyl ester 7 (3.0 g, 21.0 mmol) in 40 mL of dichloromethane was added dropwise and maintained at this temperature for half an hour. The mixture was allowed to warm to room temperature over 30 minutes. The solvent was removed in vacuo and the residue was purified eluting eluting eluting eluting eluting Confirmed by GC-MS. 4-(((lS,2S)-2-(ethyloxymethyl)cyclopropyl)butan-1,3-diynyl)benzoic acid methyl ester (1〇>

試劑 MW 當量 mmol g 5 mL 化合物8 .297.97 1 10.85 3.2 g 化合物9 160.17 1.1 15.20 2.43 g Pd2dba2 575.22 0.01 0.11 62.4 mg TMPP 352.24 0.04 0.43 153 mg NEt3 101.19 3 14.25 1.44 g DMF 50 mL 在氬氣下乙酸（（18,23)-2-(2,2-二溴乙烯基）環丙基）曱酯 8(3.2 g，10.85 mmol)、Pd2<lba2(62.4 mg，0.11 mmol)、三 (4-甲基苯基）膦（153 mg，0.43 mmol)、三乙胺（1.44 g， 159849.doc -55- 201249786 14.25 mmol)於DMF(50 mL)中之溶液用4-乙炔基苯曱酸甲酯9(2.43 g，15.2 mmol)處理。該混合物在室溫下持續攪拌隔夜。用EtOAc(300 mL)稀釋反應物，用水（3x100 mL)洗滌，乾燥經合併之有機層且濃縮，得到粗化合物10，藉由矽膠管柱層析用含乙酸乙酯之石油醚（3%至10% v/v)純化，得到呈黃色固體狀之化合物10(1.4 g，46.5%)。LC-MS:297 [M+H]+。 4-(((lS，2S)-2-(羥甲基）環丙基）丁-1,3-二炔基）苯甲酸（11)Reagent MW equivalent mmol g 5 mL Compound 8.297.97 1 10.85 3.2 g Compound 9 160.17 1.1 15.20 2.43 g Pd2dba2 575.22 0.01 0.11 62.4 mg TMPP 352.24 0.04 0.43 153 mg NEt3 101.19 3 14.25 1.44 g DMF 50 mL Acetic acid under argon ( 18,23)-2-(2,2-dibromovinyl)cyclopropyl)decyl ester 8 (3.2 g, 10.85 mmol), Pd2 <lba2 (62.4 mg, 0.11 mmol), tris(4-methylbenzene) a solution of phosphine (153 mg, 0.43 mmol), triethylamine (1.44 g, 159849.doc -55 - 201249786 14.25 mmol) in DMF (50 mL) with methyl 4-ethynylbenzoate 9 (2.43 g, 15.2 mmol). The mixture was continuously stirred at room temperature overnight. The reaction was diluted with EtOAc (EtOAc EtOAc (EtOAc)EtOAc. Purification of 10% v/v) gave compound 10 (1.4 g, 46.5%). LC-MS: 297 [M+H]+. 4-(((lS,2S)-2-(Hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzoic acid (11)

試劑 MW 當量 mmol g，mL 化合物10 296.32 1 4.72 1.4 g NaOH 40.10 4 18.90 756 mg THF 30 mL 向4-(((lS，2S)-2-(乙醢氧基曱基）環丙基）丁-1，3-二炔基）苯曱酸曱酯10(1.4 g)於THF(30 mL)中之溶液中添加氫氧化納（75 mg，1 8.9 mmol)於水（10 mL)中之溶液。該混合物在室溫下持續攪拌隔夜。此後，減壓移除溶劑，用水（50 ml) 稀釋殘餘物，用1 M HC1將pH值調節至4·0，接著用 EtOAc(4x50 mL)萃取，乾燥有機層且濃縮，得到呈黃色固體狀之化合物11(1.05 g，93%)，其未經進一步純化即用於下一步驟。LC-MS:241 [M+H]+。 (S)-3-(第三丁氧羰基胺基）-2-(4-(((lS，2S)-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺基)-3·甲基丁酸甲酯（12) 159849.doc •56· a 201249786Reagent MW equivalent mmol g, mL Compound 10 296.32 1 4.72 1.4 g NaOH 40.10 4 18.90 756 mg THF 30 mL to 4-(((lS,2S)-2-(ethyloxycarbonyl)cyclopropyl)- A solution of sodium hydride (75 mg, 1 8.9 mmol) in water (10 mL) was added to a solution of 1,3-diynyl benzoic acid decyl ester 10 (1.4 g) in THF (30 mL). The mixture was continuously stirred overnight at room temperature. After the solvent was removed, the residue was evaporated,jjjjjjjjjjjjjjjjjj Compound 11 (1.05 g, 93%) was used in the next step without further purification. LC-MS: 241 [M+H]+. (S)-3-(Tertidinoxycarbonylamino)-2-(4-(((lS,2S)-2-(hydroxymethyl)cyclopropyl)butane-1,3-diynyl) Methylbenzylamino)-3·methylbutyrate (12) 159849.doc •56· a 201249786

試劑 MW 當量 mmol g，mL 化合物11 240.25 1 4.38 1.05 g Boc·二-Me_DAP 246.30 1 4.38 1.076 g HATU 390.12 1.1 4.81 1.83 g DIPEA 129.24 3 13.10 1.69 g DMF 25 mL 向4-(((lS，2S)-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲酸 11(1.05 g，4.38 mmol)、HATU(1.83 g，4.81 mmol)於 DMF(25 mL)中之溶液中添加（S)-2-胺基-3-(第三丁氧羰基胺基）-3-甲基丁酸甲酯（1.076 g，4.38 mmoL)及 DIPEA( 1.69 g，13 · 1 mmoL)。混合物在室溫下持續攪拌5小時。此後，用水（20 mL)稀釋反應物，接著用乙酸乙酯（60 mL><3)萃取。用水及鹽水洗滌經合併之有機層，接著乾燥且濃縮，得到呈黃色油狀之化合物12(1.35 g，65%)。LC-MS:469 [M+H]+。 (S)-3-胺基-2-(4-(((lS，2S)-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺基）-3-甲基丁酸甲酯堕酸鹽（13)Reagent MW Equivalent mmol g, mL Compound 11 240.25 1 4.38 1.05 g Boc·Di-Me_DAP 246.30 1 4.38 1.076 g HATU 390.12 1.1 4.81 1.83 g DIPEA 129.24 3 13.10 1.69 g DMF 25 mL to 4-(((lS,2S)- 2-(Hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzoic acid 11 (1.05 g, 4.38 mmol), HATU (1.83 g, 4.81 mmol) in DMF (25 mL) Methyl (S)-2-amino-3-(t-butoxycarbonylamino)-3-methylbutanoate (1.076 g, 4.38 mmoL) and DIPEA ( 1.69 g, 13 · 1 mmoL) were added. The mixture was kept stirring at room temperature for 5 hours. Thereafter, the reaction was diluted with water (20 mL) and then extracted with ethyl acetate (60 mL ><3>). The combined organic layers were washed with EtOAcq. LC-MS: 469 [M+H]+. (S)-3-Amino-2-(4-(((S(2S))2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide)- Methyl 3-methylbutanoate citrate (13)

試劑 MW 當量 mmol g，mL 化合物12 468.23 1 2.78 1.3 g HCl(g) CH3OH 20 mL 將（S)-3-(第三丁氧羰基胺基）-2-(4-(((lS，2S)-2-(羥甲基）環丙基）丁-1,3-二炔基）苯曱醯胺基）-3-曱基丁酸曱酯12(1.3 g，2.78 mmol)於曱醇（20 mL)中之溶液連接至HC1裝置。接著在室溫下攪拌反應物直至TLC指示起始物質完全消耗。之後，減壓濃縮溶液得到呈黃色固體狀之13(1.1 g， 159849.doc -57- 201249786 98%)。LC-MS:369 [M+H]+。W-NMR: (DMSO人 400 MHz) 5:9.04 (d, 7=6.8 Hz, 1H), 8.38 (s, 3H), 7.98 (d, 7=6.8 Hz, 2H), 7.64 (d, 7=6.4 Hz, 2H), 4.88 (d, J=6.8 Hz, 1H), 3.72 (s, 3H), 3.45-3.42 (m, 1H), 3.28-3.24 (m, 1H), 1.49-1.46 (m, 2H), 1.40 (s, 6H), 0.95-0.90 (m，2H)。Reagent MW equivalent mmol g, mL Compound 12 468.23 1 2.78 1.3 g HCl (g) CH3OH 20 mL (S)-3-(Tertidinoxycarbonylamino)-2-(4-(((lS,2S)) 2-(Hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzoguanido)-3-indolyl decanoate 12 (1.3 g, 2.78 mmol) in decyl alcohol (20 The solution in mL) is connected to the HC1 device. The reaction was then stirred at room temperature until TLC indicated complete consumption of starting material. After that, the solution was concentrated under reduced pressure to give 13 (1.1 g, 159849.doc - 57 - 201249786 98%) as a yellow solid. LC-MS: 369 [M+H]+. W-NMR: (DMSO 400 MHz) 5: 9.04 (d, 7 = 6.8 Hz, 1H), 8.38 (s, 3H), 7.98 (d, 7 = 6.8 Hz, 2H), 7.64 (d, 7 = 6.4 Hz, 2H), 4.88 (d, J=6.8 Hz, 1H), 3.72 (s, 3H), 3.45-3.42 (m, 1H), 3.28-3.24 (m, 1H), 1.49-1.46 (m, 2H) , 1.40 (s, 6H), 0.95-0.90 (m, 2H).

(S)-3-胺基-2-(4-(((lS，2S)-2-(羥甲基）環丙基）丁·二炔基）苯曱醯胺基）-3 -曱基丁酸曱酯鹽酸鹽13(1.15 g，2.8 mmol)用異丙醇（10 mL)及50%羥胺水溶液（3.8 mL，56.8 mmol ’ 20當量）處理直至LCMS顯示處理顯著完成為止。藉由逆相HPLC(梯度為含〇%至30%乙腈之水（各自含有 0.1% TFA)，歷經120分鐘）純化粗物質，且彙集所需溶離份且凍乾，得到呈三氟乙酸鹽形式之N-((S)-3-胺基-1-(羥胺基）-3-甲基-1·側氧基丁 _2-基）-4-(((lS，2S)-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺（白色固體，446 mg，0.92 mmol，33%)。質譜資料：預期值（M+1) : 370.4，觀測值：370.2 » 質子NMR (400 MHz，dmso-A): 11.20 (s，1H)， 9.22 (br s，1H)，8.55 (d，1H，J=9.6Hz)，7.99 (br s，3H)，7.89 (dd, 2H, J-2.0, 6.8Hz), 7.61 (dd, 2H, J=1.6, 6.8Hz), 4.66 (d, 1H, J=9.6Hz), 3.40 (dd, 1H, J=5.0, 11.4Hz), 3.22 (dd, 159849.doc -58- 201249786 1H, J=5.8, 11.8Hz), 1.38-1.46 (m, 2H), 1.30 (s, 3H), 1.25 (s, 1H), 0.84-0.93 (m, 2H) 〇 D.合成N-((S)-3·胺基-1-(羥胺基）_3_甲基側氧基丁 _2_ 基）-4’-(((l，2-反）-2-(羥甲基）環丙基）乙快基）聯苯_4-甲酿胺 (1-5)(S)-3-Amino-2-(4-(((lS,2S)-2-(hydroxymethyl)cyclopropyl)butan-diynyl)phenyl)amino)-3-fluorenyl Butyl butyrate hydrochloride 13 (1.15 g, 2.8 mmol) was treated with isopropanol (10 mL) and 50% aqueous hydroxylamine (3.8 mL, 56.8 <RTIgt; The crude material was purified by reverse phase HPLC (gradient for 〇% to 30% acetonitrile in water (each containing 0.1% TFA) over 120 min) and the desired fractions were combined and lyophilized to give the trifluoroacetate salt. N-((S)-3-Amino-1-(hydroxyamino)-3-methyl-1. oxaoxybutan-2-yl)-4-(((lS,2S)-2-(((lS,2S)-2-) Hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (white solid, 446 mg, 0.92 mmol, 33%). Mass spectral data: expected value (M+1): 370.4, observed: 370.2 » NMR (400 MHz, dmso-A): 11.20 (s, 1H), 9.22 (br s, 1H), 8.55 (d, 1H, J=9.6Hz), 7.99 (br s,3H), 7.89 (dd, 2H, J-2.0, 6.8Hz), 7.61 (dd, 2H, J=1.6, 6.8Hz), 4.66 (d, 1H, J= 9.6 Hz), 3.40 (dd, 1H, J=5.0, 11.4 Hz), 3.22 (dd, 159849.doc -58- 201249786 1H, J=5.8, 11.8Hz), 1.38-1.46 (m, 2H), 1.30 ( s, 3H), 1.25 (s, 1H), 0.84-0.93 (m, 2H) 〇D. Synthesis of N-((S)-3·amino-1-(hydroxylamino)_3_methyl-oxylated _2_yl)-4'-(((l,2-trans)-2-(hydroxymethyl)cyclopropyl)ethyl hexyl)biphenyl_4-cartoamine (1-5)

向圓底燒瓶中添加4'-溴聯苯-4-曱酸甲酯（2.〇 g，6.87 mmol，1.0當量），之後添加外消旋（2-反-乙炔基環丙基）甲醇（0.991 g ’ 10.30 mmol ’ 1.5 當量）溶解於 THF(20 mL)中之溶液。向此混合物中添加雙三苯膦二氣化鈀（241 mg， 0.343 mmol，0.05 當量）、碘化銅⑴（131 mg，〇 687 mmol ’ 0.1當量），接著添加三乙胺（6 87 mL，49.3 mmol， 7_18當量）。在75°C下攪拌反應物約2小時。使反應物冷卻至室溫，濾出固體，用THF沖洗。將溶液置於冷凍箱中5 天。濃縮直至乾燥。添加DCM且形成沈澱。過濾沈澱，用, 極少量DCM沖洗。TLC顯示沈澱主要含有產物（約丨5 g固體）°用含10¾ EtOAc之己院（5 mL)濕磨，吸移出溶劑， TLC顯示固體含有產物及液體移除起始物質·溴化物及一些基本雜質。再添加含10% EtOAc之己烷（5 mL)，且擾拌3〇分鐘’過濾’用含10% EtOAc之己烷（5 mL)沖洗，且乾燥固體。得到900 mg粗4'-(((1，2-反）-2-(羥甲基）環丙基）乙炔 159849.doc -59- 201249786 基）聯苯-4-曱酸甲酯。1H nmr (DMSO-Α): δ 0-82-0.85 (m， 2Η)，1.35-1.43 (m，2Η)，3.23-3.29 (m，1Η), 3.40-3.42 (m， 1H)，3.84 (s，3H)，4.65-4.67 (t，1H)，7.43-7.45 (d，2H)， 7.67-7.70 (d，2H)，7·8〇·7·82 (d，2H)，7.99-8.01 (d，2H)。Add 4'-bromobiphenyl-4-decanoate methyl ester (2. 〇g, 6.87 mmol, 1.0 eq.) to a round bottom flask, followed by the addition of racemic (2-trans-ethynylcyclopropyl)methanol ( 0.991 g '10.30 mmol '1.5 equivalents) of a solution dissolved in THF (20 mL). To this mixture was added bistriphenylphosphine dipalladium (241 mg, 0.343 mmol, 0.05 eq.), copper iodide (1) (131 mg, 〇 687 mmol '0.1 eq.) followed by triethylamine (6 87 mL, 49.3 mmol, 7_18 equivalents). The reaction was stirred at 75 ° C for about 2 hours. The reaction was cooled to room temperature and the solid was filtered and washed with EtOAc. The solution was placed in a freezer for 5 days. Concentrate until dry. DCM was added and a precipitate formed. The precipitate was filtered and rinsed with a very small amount of DCM. TLC showed that the precipitate contained mainly product (approximately 5 g of solid). Wet was triturated with a 10⁄4 EtOAc solution (5 mL), and the solvent was removed by suction. TLC showed solids containing product and liquid removed starting material bromide and some basic Impurities. A further 10% EtOAc in hexanes (5 mL) was evaporated and evaporated. This gave 900 mg of crude 4'-((1,2-trans)-2-(hydroxymethyl)cyclopropyl)acetylene 159849.doc -59- 201249786 base)biphenyl-4-decanoate. 1H nmr (DMSO-Α): δ 0-82-0.85 (m, 2Η), 1.35-1.43 (m, 2Η), 3.23-3.29 (m, 1Η), 3.40-3.42 (m, 1H), 3.84 (s ,3H),4.65-4.67 (t,1H),7.43-7.45 (d,2H), 7.67-7.70 (d,2H),7·8〇·7·82 (d,2H),7.99-8.01 (d , 2H).

將4’-(((1，2-反）-2-(羥甲基）環丙基）乙炔基）聯笨_4_曱酸曱酯（900 mg ’ 2.94 mmol，l.o 當量）溶解於甲醇（5 mL)、 DMF(2 mL)及THF(5 mL)中。在室溫下添加 1·〇 μ NaOH (4.41 mL，4.407 mmol，1.5當量）。攪拌反應物4天。濃縮反應物，以移除MeOH及THF，用6 N HC1(約5 mL)酸化pH 值至約3。用EtOAc(3x50 mL)萃取，合併有機層，用飽和 NaCl洗滌，乾燥（MgS〇4) ’過遽，濃縮。得到890 mg 4，-(((1，2-反）-2-(羥甲基）環丙基）乙炔基）聯笨-4-曱酸。 NMR (DMSO-J5): δ 0.79-0.88 (m, 2H), 1.34-1.44 (m, 2H), 3.22-3.26 (m, 1H), 3.40-3.44 (m, 1H), 7.42-7.44 (d, 2H), 7.66-7.68 (d, 2H), 7.76-7.78 (d, 2H), 7.97-7.99 (d, 2H), 8.10 (s，1H)。Dissolving 4'-(((1,2-trans)-2-(hydroxymethyl)cyclopropyl)ethynyl) hydrazin-4-yl decanoate (900 mg ' 2.94 mmol, lo equivalent) in methanol (5 mL), DMF (2 mL) and THF (5 mL). 1·〇 μ NaOH (4.41 mL, 4.407 mmol, 1.5 eq.) was added at room temperature. The reaction was stirred for 4 days. The reaction was concentrated to remove MeOH and THF and acidified to pH 3 with 6 N EtOAc (~ 5 mL). Extracted with EtOAc (3×50 mL),EtOAc. 890 mg of 4,-((1,2-trans)-2-(hydroxymethyl)cyclopropyl)ethynyl)pyr-4-pyruic acid are obtained. NMR (DMSO-J5): δ 0.79-0.88 (m, 2H), 1.34-1.44 (m, 2H), 3.22-3.26 (m, 1H), 3.40-3.44 (m, 1H), 7.42-7.44 (d, 2H), 7.66-7.68 (d, 2H), 7.76-7.78 (d, 2H), 7.97-7.99 (d, 2H), 8.10 (s, 1H).

將（S)-2-胺基-3-(第三丁氧羰基胺基）-3-甲基丁酸甲酯 159849.doc •60- 201249786 (150 mg，0.609 mmo卜 1，〇當量）及4__(((12 反）2 (羥甲基）環丙基）乙炔基）聯苯_4-曱酸（178 mg，〇·6〇9 mm〇i，i.o當里）浴解於DMF(2 mL)中。向此混合物中添加〇1?£八（0.266 mL，1.523 mmol，2.5 當量），接著添加 HATU(278 mg， 0.731 mm〇l，1.2當量）。在室溫下攪拌此混合物約48小時。使反應物於1 Μ檸檬酸與乙酸乙酯之間分配。用半飽和氣化鈉、飽和碳酸氫鈉，接著用飽和氣化鈉洗滌有機物’經硫酸鎮乾燥且蒸發至乾燥。得到370 mg粗（s)-3-(第三丁氧羰基胺基）-2-(4’-(((18,28)-2-(羥曱基）環丙基）乙炔基）聯苯-4-基曱醯胺基）-3 -甲基丁酸甲g旨。[CMS M+1預期值=521.3，觀測值=521.3。Methyl (S)-2-amino-3-(t-butoxycarbonylamino)-3-methylbutanoate 159849.doc •60- 201249786 (150 mg, 0.609 mmo, 1, 〇 equivalent) and 4__(((12-)) 2 (hydroxymethyl)cyclopropyl)ethynyl)biphenyl_4-decanoic acid (178 mg, 〇·6〇9 mm〇i, io zhongli) bath solution in DMF (2 In mL). To this mixture was added 〇1?£8 (0.266 mL, 1.523 mmol, 2.5 eq.) followed by HATU (278 mg, 0.731 mm ,l, 1.2 eq.). The mixture was stirred at room temperature for about 48 hours. The reaction was partitioned between 1 EtOAc and EtOAc. The organics were washed with sulfuric acid sodium and saturated sodium bicarbonate followed by saturated sodium sulfate. 370 mg of crude (s)-3-(t-butoxycarbonylamino)-2-(4'-((18,28)-2-(hydroxyl)cyclopropyl)ethynyl)biphenyl 4-Glysylamino)-3-methylbutyric acid. [CMS M+1 expected value = 521.3, observed value = 521.3.

將（S)-3-(第三丁氧羰基胺基）-2-(4·-(((1,2-反）-2-(羥曱基）環丙基）乙快基）聯苯-4-基曱酿胺基）-3 -甲基丁酸甲醋（粗物質）溶解於甲醇（1 mL)中。在室溫下添加含4.0 M HC1之二 °惡烧（1.2 83 mL，5.13 mmol，8.43當量）。在90分鐘之後反應完成（HPLC)。在0°C下於旋轉蒸發器上濃縮反應物。向此添加IPA(1 mL)，接著添加經胺溶液（0.804 mL，12.18 mmol，20當量）。將燒瓶置於4°C下約120小時。反應物濃縮（使反應物保持在下）成膠黏物質。向此添加水（3 mL) 及ACN(0.5 mL)。在0°C下使用TFA(3 mL)酸化。再添加水 (1 mL)及 ACN(1 mL)。藉由 RP HPLC(1” 管柱，25 mL/min，含0.1% TFA之水/ACN，在10% B下平衡）純化。 159849.doc -61 - 201249786 使用注射過濾器（2x6.5 mL)加載於1"管柱（i〇 mL/min，5% B)上。經1分鐘勻變至25 mL/min。10% B歷時15分鐘，接著經80分鐘1〇%至70% B，產物在41分鐘至48分鐘溶離合併所需溶離份，冷凍且置於冷凍乾燥器（以〇)上。得到 105 mg N-((S)·3·胺基-1-(羥胺基）·3_甲基側氧基丁 j 基）-4，_(((1，2-反）-2-(經甲基）環丙基）乙炔基）聯笨+甲酿胺 (1-5)，叮八。1^肘8馗+1預期值=422.2’觀測值=422 2。 *H NMR (OMSO-d6)： δ 0.8〇-〇<88 (m, 2H), 1.31 (s, 3H), 1.36-1.44 (m, 2H), 3.21-3.293.45 (m，1H)，4.65-4.71 (m，2H), 7.44-7.46 7.71 (d，2H)，7.79-7.81 (d，2H)，7.97-7.998.48 (d，1H)，9.22 (s，1H)，U.21 (br，1H)。 1,27 (s, 3H), (m，lH)，3.40· (d，2H)，7.68-(d，2H)，8.46- E.合成N-((S)-M羥胺基）_3·甲基_3_(甲胺基）]側氧基丁-2-基）-4-(((l，2-反）-2-(羥甲基）環丙基）丁 ·13二炔基）苯甲醯胺（1-6)及N-((S)-l-(羥胺基）_3·(2_羥基乙胺基）_夂甲基小側氧基丁-2-基）-4-(((1,2-反）_2_(羥甲基）環丙基炔基）苯甲醯胺（1-7) ’ -一(S)-3-(Tertidinoxycarbonylamino)-2-(4·-((1,2-trans)-2-(hydroxyindenyl)cyclopropyl)ethyl)biphenyl The -4-yl-branched amino)-3-methylbutyric acid methyl vinegar (crude) was dissolved in methanol (1 mL). A dimethyl alcohol (2.083 mL, 5.13 mmol, 8.43 eq.) containing 4.0 M HCl was added at room temperature. The reaction was completed after 90 minutes (HPLC). The reaction was concentrated on a rotary evaporator at 0 °C. IPA (1 mL) was added, followed by an amine solution (0.804 mL, 12.18 mmol, 20 eq.). The flask was placed at 4 ° C for about 120 hours. The reactants are concentrated (to keep the reactants under) into a sticky substance. Water (3 mL) and ACN (0.5 mL) were added thereto. Acidified using TFA (3 mL) at 0 °C. Additional water (1 mL) and ACN (1 mL) were added. Purified by RP HPLC (1" column, 25 mL/min, water containing 0.1% TFA/ACN, equilibrated at 10% B. 159849.doc -61 - 201249786 using a syringe filter (2x6.5 mL) Load on 1"column (i〇mL/min, 5% B). Change to 25 mL/min over 1 minute. 10% B for 15 minutes, followed by 80% 1% to 70% B, product The desired fractions were combined by dissolving in 41 minutes to 48 minutes, frozen and placed in a freeze dryer (〇) to give 105 mg of N-((S)·3·amino-1-(hydroxylamine)·3_ Methyl-terminated keto)-4,-((1,2-trans)-2-(methyl)cyclopropyl)ethynyl)biphenyl+cartoamine (1-5),叮VIII.1 elbow 8馗+1 expected value=422.2' observation value=422 2. *H NMR (OMSO-d6): δ 0.8〇-〇<88 (m, 2H), 1.31 (s, 3H), 1.36-1.44 (m, 2H), 3.21-3.293.45 (m,1H), 4.65-4.71 (m,2H), 7.44-7.46 7.71 (d,2H),7.79-7.81 (d,2H),7.97- 7.998.48 (d,1H), 9.22 (s,1H), U.21 (br,1H). 1,27 (s, 3H), (m,lH), 3.40· (d,2H), 7.68- (d, 2H), 8.46- E. Synthesis of N-((S)-Mhydroxylamino)_3·methyl_3_(methylamino)] oxobutan-2-yl)-4-(((l 2-trans)-2-(hydroxymethyl)cyclopropyl)butyl·13 diynyl)benzamide (1-6) and N-((S)-l-(hydroxylamino)_3·(2 _Hydroxyethylamino)-夂methyl-small oxybutan-2-yl)-4-(((1,2-trans)_2-(hydroxymethyl)cyclopropylalkynyl)benzamide (1 -7) ' - one

159849.doc 62· 201249786 於250 mL圓底燒瓶中添加含N-((S)-3-胺基-1-(羥胺基）-3-甲基-1·•侧氧基丁-2-基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁-1,3-二炔基）苯甲醯胺（1-1)(1.2 g ’ 3.25 mmol)、三乙胺（2 mL，14.35 mmol)及甲搭（0.2 g，6.66 mmol)之 DMF(20 ml)，得到黃色溶液。攪拌反應混合物3小時。用正丁胺（2 ml)淬滅過量甲醒·。在室溫下在〇°C下添加氰基棚氫化納 (570 mg，14.3 mmol)及乙酸（4 ml)。藉由LCMS檢查反應混合物。移除溶劑之後，藉由HPLC純化產物，得到440 mg N-((S)-l-(羥胺基）-3-曱基-3-(曱胺基）-1-侧氧基丁-2-基）-4-(((1,2-反）-2-(羥曱基）環丙基）丁-1，3-二炔基）苯甲醯胺 1-6。LC-MS (M+1) 384 ;化學式：C2丨H25N304，精確質量：383.18。NMR (DMSO-A)，TFA鹽：δ 0.88 (m，2H)， 1.30 (s，3H)，1.35 (s，3H)，1.44 (m，1H)，2.51 (s，H)，3.39 (m，1H)，3.41 (m，1H)，4.7( br.d，1H)，4.84 (d，1H)，7.58 (d， 2H), 7.91 (d, 2H), 8.51 (br.m, 1NH) 8.64 (d, 1NH), 9.23 (s, NH)，11.17 (s，OH)。於250 mL圓底燒瓶中添加含N-((S)-3-胺基-1-(羥胺基）-3-曱基-1-側氧基丁-2-基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺（1-1)(1 g，2.71 mmol)、三乙胺（0.377 mL，2.71 mmol)及乙越（0.119 g，2.71 mmol)之 DMF(20 ml)，得到黃色溶液。揽拌反應混合物3小時。在室溫下添加氰基棚氫化鈉（570 mg，14.3 mmol)及乙酸（2 ml)。藉由 LCMS檢查反應混合物。濃縮反應混合物且藉由jjPLC純化產物’得到642 mg N-((S)-l-(經胺基）-3-(2-經基乙胺基）-3- 159849.doc •63- 201249786 甲基-1-侧氧基丁-2-基）-4-(((1,2-反）-2-(羥曱基）環丙基）丁-1，3-二炔基）苯曱醯胺1-7。LC-MS (M+1) 398;化學式： C22H27N304，精確質量：397.20 » 4 NMR (DMSO-A)， TFA鹽：δ 0.85 (m，2H)，1.14(t，3H)，1.29 (s，3H)，1.39 (s， 3H), 1.44 (m, 1H), 2.96(br.m, 2H), 3.24 (m, 2H), 3.39 (m, 2H), 4.64(br.d, 1H), 4.85 (d, 1H), 7.60 (d, 2H), 7.92 (d, 2H), 8.59 (br.m, 1NH), 8.45(brt, 1NH), 9.23 (s, NH), 11.15 (s，OH) » F.合成N-((S)-l-(羥胺基）-3-(2-羥基乙胺基）-3-甲基-1-側氧基丁 -2-基）-4-(((1,2-反）-2-(經甲基）環丙基）丁 _ι，3-二炔基）苯甲醯胺（1-8)159849.doc 62· 201249786 Adding N-((S)-3-amino-1-(hydroxylamino)-3-methyl-1·•oxybutan-2-yl group to a 250 mL round bottom flask )-4-(((1,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (1-1) (1.2 g ' 3.25 mmol) Triethylamine (2 mL, 14.35 mmol) and M.p. (0.2 g, 6.66 mmol) of DMF (20 ml). The reaction mixture was stirred for 3 hours. Excessive awakening was quenched with n-butylamine (2 ml). Cyanide hydride (570 mg, 14.3 mmol) and acetic acid (4 ml) were added at room temperature under 〇 °C. The reaction mixture was checked by LCMS. After removing the solvent, the product was purified by HPLC to give 440 mg of N-((S)-l-(hydroxylamino)-3-indolyl-3-(decylamino)-1-oxetet-2- 4-((1,2-trans)-2-(hydroxyindenyl)cyclopropyl)butan-1,3-diynyl)benzamide 1-6. LC-MS (M+1) 384; Chemical formula: C2 丨H25N304, exact mass: 383.18. NMR (DMSO-A), TFA salt: δ 0.88 (m, 2H), 1.30 (s, 3H), 1.35 (s, 3H), 1.44 (m, 1H), 2.51 (s, H), 3.39 (m, 1H), 3.41 (m, 1H), 4.7 (br.d, 1H), 4.84 (d, 1H), 7.58 (d, 2H), 7.91 (d, 2H), 8.51 (br.m, 1NH) 8.64 ( d, 1NH), 9.23 (s, NH), 11.17 (s, OH). Adding N-((S)-3-amino-1-(hydroxyamino)-3-indol-1-yloxybutan-2-yl)-4-((() in a 250 mL round bottom flask 1,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (1-1) (1 g, 2.71 mmol), triethylamine (0.377 M, 2.71 mmol) and EtOAc (0.119 g, 2. The reaction mixture was stirred for 3 hours. Sodium cyanohydride (570 mg, 14.3 mmol) and acetic acid (2 ml) were added at room temperature. The reaction mixture was checked by LCMS. The reaction mixture was concentrated and the product was purified by jjPLC to give 642 mg of N-((S)-l-(amino)-3-(2-ethylethylamino)-3- 159849.doc •63- 201249786 Keto-1-oxobutan-2-yl)-4-(((1,2-trans)-2-(hydroxyindenyl)cyclopropyl)butan-1,3-diynyl)phenylhydrazine Amine 1-7. LC-MS (M+1) 398; Chemical formula: C22H27N304, exact mass: 397.20 » 4 NMR (DMSO-A), TFA salt: δ 0.85 (m, 2H), 1.14 (t, 3H), 1.29 (s, 3H) ), 1.39 (s, 3H), 1.44 (m, 1H), 2.96 (br.m, 2H), 3.24 (m, 2H), 3.39 (m, 2H), 4.64 (br.d, 1H), 4.85 ( d, 1H), 7.60 (d, 2H), 7.92 (d, 2H), 8.59 (br.m, 1NH), 8.45 (brt, 1NH), 9.23 (s, NH), 11.15 (s,OH) » F Synthesis of N-((S)-l-(hydroxylamino)-3-(2-hydroxyethylamino)-3-methyl-1-oxobutan-2-yl)-4-(((1) ,2-trans)-2-(methyl)cyclopropyl)butyrate,3-diynyl)benzamide (1-8)

於250 mL圓底燒瓶（t=g)中添加含N-((S)-3-胺基-1-(經胺基）-3 -甲基-1-側氧基丁-2-基）-4-(((1,2-反）-2-(經甲基）環丙基）丁-1，3-二炔基）苯甲醯胺（1 g，2_71 mmol)及三乙胺 (0.3 77 ml ’ 2.71 mmol)之DMF(20 ml)，得到黃色溶液。添加2-(第二丁基一甲基梦烧基氧基）乙越（0.494 g，2 83 mmol)。攪拌反應混合物1 8小時，濃縮至乾燥。將殘餘物 159849.doc -64 - 201249786 溶解於THF(15 ml)中且在室溫下添加乙酸（2 ml)及氰基硼氫化鈉（600 mg，15 mmol)。藉由lcMS檢查反應混合物。在室溫下攪拌2小時之後，添加TFA(5 ml)且再攪拌3小時。由LCMS確認反應完成，且濃縮反應物，且經HpLc純化產物，得到370 mg N-((S)-l-(羥胺基）_3·(2·羥基乙胺基 3-曱基-1-側氧基丁-2-基）-4-((( 1,2-反）_2_(羥甲基）環丙基）丁-1，3 -— 炔基）苯甲醯胺 1-8。LC-MS (M+1) 414 ;化學式：C22H27N305，精確質量：413.20 ; 4 NMR (DMSO-⑹，TFA鹽：δ 0.88 (m, 2H)，1.30 (s, 3H)，1.35 (s，3H)，1.44 (m, 1H), 3.22 (m, 2H), 3.39 (m, 2H), 4.64(br.d, 1H), 4.90 (d, 1H), 5.25(br. S, 1H), 7.60 (d, 2H), 7.91 (d, 2H), 8.59 (br.m，1NH)，8.71 (d，1NH)，9.26 (s，NH)，11.19 (s, OH)。 G.合成N-((S)-3-(二甲胺基）-1-(羥胺基）_3_甲基•侧氧基丁 _2·基）-4-(((1，2_反）·2·(羥甲基）環丙基）丁_13_二炔基）苯甲醯胺（1-9)Add N-((S)-3-amino-1-(amino)-3-methyl-1-oxobutan-2-yl) to a 250 mL round bottom flask (t=g) 4-((1,2-trans)-2-(methyl)cyclopropyl)butan-1,3-diynyl)benzamide (1 g, 2_71 mmol) and triethylamine ( 0.377 ml ' 2.71 mmol) of DMF (20 ml) gave a yellow solution. 2-(Second-butyl-methylmethyloxyalkyloxy) acetophenone (0.494 g, 2 83 mmol) was added. The reaction mixture was stirred for 18 h and concentrated to dryness. The residue 159849.doc -64 - 201249786 was dissolved in THF (15 ml) and acetic acid (2 ml) and sodium cyanoborohydride (600 mg, 15 mmol) were added at room temperature. The reaction mixture was checked by lcMS. After stirring at room temperature for 2 hours, TFA (5 ml) was added and stirred for additional 3 hours. The reaction was confirmed to be complete by LCMS, and the reaction was concentrated, and the product was purified by HpLc to give 370 mg of N-((S)-l-(hydroxyamino)-3·(2·hydroxyethylamino 3-mercapto-1-one Oxybutan-2-yl)-4-(((1,2-trans)_2-(hydroxymethyl)cyclopropyl)butan-1,3--ynyl)benzamide 1-8. LC- MS (M+1) 414; Chemical formula: C22H27N305, exact mass: 413.20; 4 NMR (DMSO-(6), TFA salt: δ 0.88 (m, 2H), 1.30 (s, 3H), 1.35 (s, 3H), 1.44 (m, 1H), 3.22 (m, 2H), 3.39 (m, 2H), 4.64 (br.d, 1H), 4.90 (d, 1H), 5.25 (br. S, 1H), 7.60 (d, 2H ), 7.91 (d, 2H), 8.59 (br.m, 1NH), 8.71 (d, 1NH), 9.26 (s, NH), 11.19 (s, OH). G. Synthesis of N-((S)-3 -(dimethylamino)-1-(hydroxylamino)_3_methyl•peroxybutan-2-yl)-4-(((1,2_trans)·2·(hydroxymethyl)cyclopropane )) _13_diynyl) benzamide (1-9)

將N-((S)-3-胺基-1-(羥胺基）-3-甲基-1-側氧基丁 _2_基）_4_ ((2-(羥曱基）環丙基）丁 _ι，3·二炔基）笨甲醯胺（3〇〇 mg， 0.81 mmol)溶解於見沁二甲基甲醯胺（1〇 mL)中且在室溫下用三聚甲醛（732 mg ’ 8.1 mmol ’ 10當量）及二異丙基乙胺（0.56 mL，3.3 mmol，4當量）處理16小時。引入三氟 159849.doc -65· 201249786 乙酸（1.3 mL，16.2 mmol，2〇當量）及氰基硼氩化鈉（1〇1 mg，1.6 mmol，2 當量）。藉由逆相HPLC純化粗物質，且彙集所需溶離份且凍乾，得到呈三氟乙酸鹽形式之N_((s)_3_(二曱胺基）_卜（羥胺基）-3_曱基-1-側氧基丁-2-基）-4-(((1，2-反）-2-(羥甲基）環丙基）丁 -1，3-二炔基）苯曱醯胺（白色固體，29 mg，5.3 μηιοί，0.7%)。質譜資料：預期值（M+1) · 398 5，觀測值：398.2。質子NMR (400 MHz，dms〇〇: ll.16 (s，1H)， 9.25 (br s, 1H), 9.05 (br s, 1H), 8.68 (d3 1H, J=l〇.〇 Hz), 7.94 (dd, 2H, 1=1.6, 8.0 HZ), 7.59 (dd, 2H, 1=1.8, 8.2 Hz), 5.06 (d，1H，J=9.6 Hz)，3.4 (峰經水模糊），3 21 (dd，1H， 1=5.2, 11.2 Hz), 2.75 (d, 3H, J=5.6 Hz), 2.73 (d, 3H, J=5.2N-((S)-3-Amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)_4_((2-(hydroxyindenyl)cyclopropyl)丁_ι,3·diynyl) benzoic acid (3 〇〇 mg, 0.81 mmol) was dissolved in dimethyl dimethyl carbamide (1 〇 mL) and treated with trioxane at room temperature (732 Mp '8.1 mmol '10 equivalents) and diisopropylethylamine (0.56 mL, 3.3 mmol, 4 eq.) were treated for 16 hours. Introduction of trifluoro 159849.doc -65· 201249786 acetic acid (1.3 mL, 16.2 mmol, 2 〇 equivalent) and sodium cyanoborohydride (1 〇 1 mg, 1.6 mmol, 2 eq.). The crude material was purified by reverse phase HPLC, and the desired fractions were combined and lyophilized to give N-((s)_3_(didecylamino)-b (hydroxylamino)-3-fluorenyl in the form of trifluoroacetate. -1-Sideoxybutan-2-yl)-4-(((1,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (white solid, 29 mg, 5.3 μηιοί, 0.7%). Mass spectral data: expected value (M+1) · 398 5, observed: 398.2. Proton NMR (400 MHz, dms 〇〇: ll.16 (s, 1H), 9.25 (br s, 1H), 9.05 (br s, 1H), 8.68 (d3 1H, J=l〇.〇Hz), 7.94 (dd, 2H, 1=1.6, 8.0 HZ), 7.59 (dd, 2H, 1=1.8, 8.2 Hz), 5.06 (d, 1H, J=9.6 Hz), 3.4 (peak water blurred), 3 21 ( Dd,1H, 1=5.2, 11.2 Hz), 2.75 (d, 3H, J=5.6 Hz), 2.73 (d, 3H, J=5.2

Hz), 1.48 (s, 3H), 1.39-1.45 (m, 2H), 1.28 (s, 3H), 0.85-0.91 (m, 2H)。 H.合成N-((S)-3-羥基羥胺基广3_甲基q側氧基丁 2_ 基）-4-(((1，2-反）_2·(經甲基）環丙基）丁-1，3-二炔基）苯甲醯胺（1-10)Hz), 1.48 (s, 3H), 1.39-1.45 (m, 2H), 1.28 (s, 3H), 0.85-0.91 (m, 2H). H. Synthesis of N-((S)-3-hydroxyhydroxylamino)3(methylq-oxyl-2-yl)-4-(((1,2-trans)_2·(methyl)cyclopropyl )-1,3-1,3-diynyl)benzamide (1-10)

在室溫下向攪拌之4-(((1，2·反）-2-(羥甲基）環丙基）丁_ 1,3-二炔基）苯甲酸（75〇 mg ’ 3 12 、HATU(570 mg， 159849.doc ·66· 201249786 1·5 mmol)及 DIEA(3.4 ml，過量）於DMF(30 ml)中之溶液中添加（S)-2-胺基-3-羥基-3-曱基丁酸曱酯鹽酸鹽（402 mg， 1.5 mmol)。在環境溫度下攪拌反應混合物1小時，之後用水（5〇 ml)稀釋。用乙酸乙酯（1〇〇 ml><3)及鹽水（20 ml)萃取溶液。經MgS04乾燥有機層且蒸發。將所得產物（1.3 g)溶解於IPA(20 mL)中且用NH2OH(10 mL，過量）處理，且在室溫下攪拌3天。移除過量溶劑，且經逆相HPLC純化粗產物’得到178 mg N-((S)-3-羥基-1-(羥胺基）-3-曱基-i_側氧基丁-2-基）-4-(((1，2-反）-2-(羥曱基）環丙基）丁-l，3-二炔基）苯甲醯胺 1-10 ; LC-MS (M+1) 371 ;化學式：C20H22N2O5，精確質量：370.15。4 NMR (DMSO-A): δ 0.88 (m, 2H)， 1.11 (s, 3H), 1.16 (s, 3H), 1.44 (m, 1H), 3.20 (m, 1H), 3.40 (m，2H)，4.34(d，1H), 4.84 (d, 1H)，7·58 (d，2H)，7.84 (d， 2H)，7.6 (d，1NH)，8.88(br.s，NH)，10.57 (s, OH)。 I·合成N-((S)-3 -胺基-1-(經胺基）-3-甲基-1·側氧基丁 _2_ 基）_4-(((lS，2R)-2-(羥甲基）-2_甲基環丙基）丁·二炔基）苯甲醯胺（1-11)Stirring 4-(((1,2·trans)-2-(hydroxymethyl)cyclopropyl)butyla-1,3-diynyl)benzoic acid (75〇mg ' 3 12 , Add (S)-2-amino-3-hydroxy-3 to a solution of HATU (570 mg, 159849.doc ·66· 201249786 1.5 mmol) and DIEA (3.4 ml, excess) in DMF (30 ml) - mercapto decyl octoate hydrochloride (402 mg, 1.5 mmol). The reaction mixture was stirred at ambient temperature for 1 hour then diluted with water (5 mL). ethyl acetate (1 〇〇ml <<3> The solution was extracted with brine (20 ml). The organic layer was dried with EtOAc EtOAc.jjjjjjjjjjjjjjjjjjjjj 3 days. Excess solvent was removed and the crude product was purified by reverse phase HPLC to give 178 mg of N-((S)-3-hydroxy-1-(hydroxylamino)-3-indolyl-i-trioxybutene- 2-yl)-4-(((1,2-trans)-2-(hydroxyindenyl)cyclopropyl)butan-l,3-diynyl)benzamide 1-10 ; LC-MS ( M+1) 371; chemical formula: C20H22N2O5, exact mass: 370.15. 4 NMR (DMSO-A): δ 0.88 (m, 2H), 1.11 (s, 3H), 1.16 (s, 3H), 1.44 (m, 1H) ), 3.20 (m, 1H), 3.40 (m, 2H), 4.34 (d, 1H), 4.84 (d, 1H), 7·58 (d, 2H), 7.84 (d, 2H), 7.6 (d, 1NH), 8.88 (br.s, NH), 10.57 (s, OH). I·Synthesis of N-((S)-3-amino-1-(amino)-3-methyl-1· oxetyl-2-yl)_4- (((lS,2R)-2-(Hydroxymethyl)-2_methylcyclopropyl)butanenyl)benzamide (1-11)

159849.doc •67· 201249786159849.doc •67· 201249786

ACHLOWI2·中間物·Α (S，E)-3-(2,2-二甲基-1,3-二氧雜環戍烷-4-基）-2-甲基丙烯酸乙酯（3)ACHLOWI2·Intermediate·Α(S,E)-3-(2,2-Dimethyl-1,3-dioxan-4-yl)-2-methylpropenoic acid ethyl ester (3)

試劑 MW 當量 mmol g，mL 化合物1 262.30 1 49.6 13.0 g NaI04 213.89 1.2 59.5 12.7 g 化合物2 238.22 2 99.2 23.0 g K2C〇3 138.15 2.1 103.6 14.3 g NaHC03(5% 水溶液） 60 mL 在室溫下向攪拌之二丙酮D-甘露糖醇1(13.0 g，49.6 mmol)於碳酸氫鈉（5%水溶液，60 ml)中之溶液中逐滴添加過硤酸納（12.74 g，59.5 mmo 1)之飽和溶液。添加後，混合物在室溫下持續攪拌2小時。接著添加2-(二乙氧基磷醯基）丙酸乙酯2(23.0 g，99.2 mmol)，之後添加碳酸lf(14.3 g， 103.6 mmol)，混合物在室溫下再持續攪拌120小時。將混合物添加至500 ml水中，用EtOAc(300 mL><3)萃取。經無水硫酸鈉乾燥經合併之有機層，濃縮且接著用PE:EA (30/1)層析，得到5.5 g呈無色油狀之所需化合物3，產率： 51.8%。 (S，E)-3-(2,2-二甲基-1,3-二氧雜環戍烷-4-基）-2-甲基丙-2-0-1-0(4) 159849.doc -68- β 201249786Reagent MW equivalent mmol g, mL Compound 1 262.30 1 49.6 13.0 g NaI04 213.89 1.2 59.5 12.7 g Compound 2 238.22 2 99.2 23.0 g K2C〇3 138.15 2.1 103.6 14.3 g NaHC03 (5% aqueous solution) 60 mL Stirring at room temperature A saturated solution of sodium perrhenate (12.74 g, 59.5 mmo 1) was added dropwise to a solution of diacetone D-mannitol 1 (13.0 g, 49.6 mmol) in sodium hydrogencarbonate (5% aqueous solution, 60 ml). After the addition, the mixture was continuously stirred at room temperature for 2 hours. Ethyl 2-(diethoxyphosphonio)propanoate 2 (23.0 g, 99.2 mmol) was then added followed by EtOAc (14.3 g, 103.6 mmol). The mixture was added to 500 ml of water and extracted with EtOAc (300 mL ><3). The combined organic layers were dried with EtOAc EtOAc m. (S,E)-3-(2,2-dimethyl-1,3-dioxan-4-yl)-2-methylpropan-2-0-1-0(4) 159849 .doc -68- β 201249786

試劑 MW 當量 mmol g » mL 化合物3 214.26 1 25 5.35 g DIBALH THF 1Μ之甲苯溶液 2 50 50 mL 150 mL 在氬氣下在ot：下向（S，E)-3-(2,2-二甲基-1，3_二氧雜環戊烷-4-基）-2-甲基丙烯酸乙酯3(5 35 g，5 mm〇1)Kl5〇⑹ THF中之溶液中逐滴添加DIBAL_H(5〇 mL，5〇 mm〇^之溶液。添加後，混合物在Ot下持續攪拌3小時。當1^(：：顯示剩餘少量化合物3時，洧30 ml氨水淬滅反應，且與3〇〇 ml 一氯甲院一起再挽拌1小時。過濾混合物，且用THF洗務殘餘物，乾燥經合併之有機層且濃縮，得到粗物質4，藉由矽膠管柱用PE:EA=10:1至5:1純化，獲得3·0 g呈無色油狀之化合物4，產率：70%。乙酸（S，E)-3-(2,2-二甲基-1，3-二氧雜環戍烷-4-基）-2-甲基稀丙酯（5)Reagent MW equivalent mmol g » mL Compound 3 214.26 1 25 5.35 g DIBALH THF 1 Torr in toluene solution 2 50 50 mL 150 mL Under argon at ot: down (S, E)-3-(2,2-dimethyl Addition of DIBAL_H (5) to a solution of ethyl-1,3-dioxol-4-yl)-2-methyl acrylate 3 (5 35 g, 5 mm 〇 1) Kl 5 〇 (6) in THF 〇mL, 5〇mm〇^ solution. After the addition, the mixture was continuously stirred for 3 hours under Ot. When 1^(:: showed a small amount of compound 3 remaining, 洧30 ml of ammonia water was quenched and reacted with 3 〇〇ml The mixture was again stirred for 1 hour. The mixture was filtered, and the residue was washed with THF. The combined organic layer was dried and concentrated to afford crude material 4 with EtOAc: Purification of 5:1 to obtain 1.00 g of Compound 4 as a colorless oil, yield: 70%. (S,E)-3-(2,2-dimethyl-1,3-dioxe. Decane-4-yl)-2-methyl-propyl propyl ester (5)

試劑 MW 當量 mmol g，mL 化合物4 172.22 1 17 2.92 g NEt3 101.19 2.5 42.5 4.3 g AcCl 78.5 2 34 2.7 g THF 100 mL 向（S，E)-3-(2,2-二甲基_1，3_二氧雜環戊烷_4_基）_2_甲基丙-2-烯-1-醇 4(2.92 g，17 mmol)於 1〇〇 ml THF 中之溶液中添加三乙胺（4.3 g，42.5 mmol)。接著在氬氣下在〇。〇下逐滴添加乙醯氯（2.7 g ’ 34 mmol)。添加後，混合物在〇。〇下持續攪拌3小時。當TLC顯示剩餘少量化合物4時，將反應物傾入150 ml水中。用EtOAc(50 mLx3)萃取。經無水硫酸 159849.doc •69- 201249786 鈉乾燥經合併之有機層，濃縮且接著用PE:EA(10/1)層析，得到3.4 g呈無色油狀之所需化合物5，產率：89%。乙酸（（反h2-((S)-2,2-二甲基-l，3-二氧雜環戊烷-4-基μΐ-甲基環丙基）甲酯（6)Reagent MW equivalent mmol g, mL Compound 4 172.22 1 17 2.92 g NEt3 101.19 2.5 42.5 4.3 g AcCl 78.5 2 34 2.7 g THF 100 mL to (S,E)-3-(2,2-dimethyl-1,3 Add triethylamine (4.3 g) to a solution of 1 mM THF in THF (4. 2 g, 17 mmol) , 42.5 mmol). Then it was under argon. Ethyl chloride (2.7 g '34 mmol) was added dropwise under the arm. After the addition, the mixture is in the mash. Keep stirring for 3 hours under the armpit. When TLC showed a small amount of Compound 4 remaining, the reaction was poured into 150 ml of water. Extract with EtOAc (50 mL x 3). The combined organic layers were dried over anhydrous EtOAc EtOAc EtOAc EtOAc EtOAc EtOAc. %. Acetic acid ((anti-h2-((S)-2,2-dimethyl-l,3-dioxolan-4-yl)-methylcyclopropyl)methyl ester (6)

試劑 MW 當量 mol g，mL 化合物5 214.26 1 15 3-2 g EtjZn 1Μ己烷溶液 5 75 75 mL ch2i2 267.84 10 150 40 g 己烷 200 mLReagent MW equivalent mol g,mL Compound 5 214.26 1 15 3-2 g EtjZn 1 hexane solution 5 75 75 mL ch2i2 267.84 10 150 40 g hexane 200 mL

在氬氣下在-20°C下向乙酸（S，E)-3-(2,2-二甲基-1，3-二氧雜環戊烧-4-基）-2-甲基稀丙酯5(3.2 g，15 mmol)於200 mL 己烷中之溶液中逐份添加二乙基辞（1 Μ己烷溶液，75 mL) »添加之後，低於-2(TC下在強烈攪拌下逐滴添加二碘甲烷（40 g，150 mol)。混合物在低於_2〇。〇下再持續攪拌8 小時，且使其升溫至室溫，且再持續攪拌8小時。藉由1〇〇 ml冷ΝΗπΐ水溶液淬滅反應混合物，接著用***（丨〇〇 mlx5)萃取。用硫代硫酸鈉水溶液、水、鹽水洗滌經合併之有機層，接著乾燥且濃縮，得到粗物質6，藉由矽膠管柱（PE:EA=3〇: i至20:1)純化得到3 〇 g呈無色油狀之“產率：88%。乙酸(（反二窥乙基•甲基環丙基）甲_⑺(S,E)-3-(2,2-dimethyl-1,3-dioxol-4-yl)-2-methyl dilute acetate at -20 ° C under argon Add propyl (1 Μ hexane solution, 75 mL) to a solution of propyl ester 5 (3.2 g, 15 mmol) in 200 mL of hexanes » after addition, below -2 (strong stirring under TC) Diiodomethane (40 g, 150 mol) was added dropwise. The mixture was stirred at less than _2 Torr for an additional 8 hours, and allowed to warm to room temperature, and stirring was continued for another 8 hours. The reaction mixture was quenched with EtOAc EtOAc EtOAc (EtOAc m. Purification of the ruthenium tube column (PE: EA = 3 〇: i to 20:1) gave 3 〇g as a colorless oil. Yield: 88%. Acetic acid ((anti-p-ethylethyl) _(7)

試劑 --一-MW 當量 mol q » mT 化合物6 AcOH(80%) 228.28 1 13 2-97 g ' 30 mL 乙鲅((反)-2-((S)-2,2-二甲基]，3二氧雜環戊烷_4·基w 159849.doc •70· 201249786 甲基環丙基）甲酯6(2.97 g，13 mmol)於30 ml 80%乙酸中之混合物在室溫下持續攪拌15小時。當TLC顯示剩餘少量6 時，將混合物逐滴添加至300 ml飽和碳酸氫鈉水溶液中。接著用二氣甲烷（200 mlx3)萃取。用水、鹽水洗滌經合併之有機層，乾燥且濃縮，得到粗物質7，藉由矽膠管柱 (PE:EA=3:1至1:1)純化得到1.5 g呈無色油狀之化合物7。產率：62%。己酸（（反）-2-甲醯基-1-甲基環丙基）γ酯（8)Reagent - one-MW equivalent mol q » mT compound 6 AcOH (80%) 228.28 1 13 2-97 g ' 30 mL acetamidine ((trans)-2-((S)-2,2-dimethyl) ,3 dioxolane-4 radical w 159849.doc •70· 201249786 methylcyclopropyl)methyl ester 6 (2.97 g, 13 mmol) in 30 ml 80% acetic acid mixture at room temperature Stir for 15 hours. When TLC showed a small amount of 6, the mixture was added dropwise to 300 mL of saturated aqueous sodium bicarbonate, then extracted with di-methane (200 ml×3). The combined organic layers were washed with water and brine and dried. Concentration to give the crude material (yield: EtOAc = EtOAc = EtOAc = EtOAc: 2-Mercapto-1-methylcyclopropyl) γ ester (8)

試劑 MW 當量 mmol g，mL 化合物7 188.22 1 7.5 1.41 g NaI04 213.89 1.2 9 1.93 g NaHC03(5% 水溶液） 20 mL 在室溫下向攪拌之乙酸（（反）-2-((S)-1,2-二羥乙基）-1-甲基環丙基）曱酯7(1.41 g，7.5 mmol)於20 ml碳酸氫鈉（5%水溶液）中之溶液中逐滴添加過埃酸鈉（1.93 g，9 mmol)飽和溶液。添加後，混合物在室溫下持續攪拌2小時。當TLC 顯示剩餘少量7時，將混合物添加至50 ml水中，用 EtOAc( 100 mLX 3)萃取。經無水硫酸納乾燥經合併之有機層，濃縮且接著用PE:EA(20/1)層析，得到1.0 g呈無色油狀之所需化合物8，產率：85%。 ((反）-2-乙炔基·1·甲基環丙基）甲醇(9)Reagent MW equivalent mmol g, mL Compound 7 188.22 1 7.5 1.41 g NaI04 213.89 1.2 9 1.93 g NaHC03 (5% aqueous solution) 20 mL Stirred acetic acid at room temperature ((trans)-2-((S)-1, 2-Dihydroxyethyl)-1-methylcyclopropyl)decyl ester 7 (1.41 g, 7.5 mmol) was added dropwise to a solution of 20 ml of sodium hydrogencarbonate (5% aqueous solution). g, 9 mmol) saturated solution. After the addition, the mixture was continuously stirred at room temperature for 2 hours. When TLC showed a small amount of 7 remaining, mixture was added to 50 ml of water and extracted with EtOAc (100 mL EtOAc). The combined organic layers were dried <RTI ID=0.0> ((trans)-2-ethynyl·1·methylcyclopropyl)methanol (9)

試劑 MW 當量 mmol g，mL 化合物8 156.08 1 5 780 mg 貝斯特曼試劑(Bestmann reagent) 192.11 1.5 7.5 1.44 g K2C〇3 138.46 3 15 2.07 g CH3OH 15 mL 159849.doc -71 - 201249786 向乙酸（（反）-2-甲醯基_ι_曱基環丙基）甲酯8(78〇，5 mmol)於15 mL甲醇中之溶液中添加貝斯特曼試劑（i 44 g， 7.51^〇1)及碳酸鉀（2.078’15111111〇1)，混合物在室溫下持續撥拌5小時。當TLC顯不剩餘少量8時，用水（2〇 mL)稀釋反應混合物。用***（20 mLx3)萃取，乾燥經合併之有機層且濃縮，得到粗物質9，藉由矽膠管柱（PE:Et2O=10:l至 5:1)純化，得到400 mg呈無色油狀之9。產率：73%。 4-(((反）-2-(羥甲基）-2-甲基環丙基）τ_13_二炔基）苯甲酸 f 酯（11)Reagent MW equivalent mmol g, mL Compound 8 156.08 1 5 780 mg Bestmann reagent 192.11 1.5 7.5 1.44 g K2C〇3 138.46 3 15 2.07 g CH3OH 15 mL 159849.doc -71 - 201249786 To acetic acid ((reverse) Adding a betmann reagent (i 44 g, 7.51^〇1) to a solution of 2-methylindolyl_ι_merylcyclopropyl)methyl ester 8 (78〇, 5 mmol) in 15 mL of methanol Potassium carbonate (2.078'15111111〇1), the mixture was continuously stirred at room temperature for 5 hours. When TLC showed no small amount of 8 remaining, the reaction mixture was diluted with water (2 mL). The mixture was extracted with diethyl ether (20 mL×3), EtOAcjjjjjjjjjjjjjjjjjjjjjjjjjj 9. Yield: 73%. 4-(((trans)-2-(hydroxymethyl)-2-methylcyclopropyl)τ_13-diynyl)benzoic acid f ester (11)

試劑 MW 當量 mmol g，mL 化合物9 110.17 1 1 3.6 400 mg 化合物10 239.07 1 1 3.6 870 mg Pd(PPh3)2Cl2 700.13 0.05 0. 18 126 mg Cul 190.23 0.1 0.36 68.4 mg i-Pr2NH 101.19 3 10.8 1.09 g THF 20 mL 在氬氣下（（反）-2-乙炔基_ι_甲基環丙基）曱醇9(4〇〇 mg， 3·6 mmol)、Pd(PPh3)2Cl2(126 mg，0.18 mmol)、Cul(68.4 mg，0.36 mmol)、DiPEA(1.〇9 g , 1〇 8 _〇1)於 2〇 mL THF 中之溶液中用4-(溴乙炔基）苯甲酸甲酯1〇(87〇 mg ’ 3 6 mmol)處理《混合物在室溫下持續攪拌5小時。當TLc顯示剩餘少量化合物9時，減壓移除溶劑，用水（5〇 mL)稀釋殘餘物，用二氣曱烷（3x5〇 mL)萃取，乾燥有機層且濃縮，得到呈紅色油狀之粗物質U，藉由矽膠管柱層析 (PE:EA=20:1至1〇:1)純化得到43() mg呈黃色固體狀之n。 159849.doc •72-Reagent MW equivalent mmol g, mL Compound 9 110.17 1 1 3.6 400 mg Compound 10 239.07 1 1 3.6 870 mg Pd(PPh3)2Cl2 700.13 0.05 0. 18 126 mg Cul 190.23 0.1 0.36 68.4 mg i-Pr2NH 101.19 3 10.8 1.09 g THF 20 mL under argon ((trans)-2-ethynyl_ι_methylcyclopropyl) decyl alcohol 9 (4 〇〇 mg, 3.6 mmol), Pd(PPh3) 2Cl2 (126 mg, 0.18 mmol) ), Cul (68.4 mg, 0.36 mmol), DiPEA (1. 〇9 g, 1〇8 _〇1) in a solution of 2 mL of THF with methyl 4-(bromoethynyl)benzoate 1 〇 ( 87 〇 mg ' 3 6 mmol) Treatment The mixture was stirred continuously for 5 hours at room temperature. The solvent was removed under reduced pressure. Substance U was purified by hydrazine column chromatography (PE: EA = 20:1 to 1 :1) to yield 43 () mg as a yellow solid. 159849.doc •72-

S 201249786 產率：40%。 4-(((lS，2R)-2-(羥甲基）-2-甲基環丙基）丁·^-二炔基）笨甲酸(ACHL02-112·中間物-A)S 201249786 Yield: 40%. 4-(((lS, 2R)-2-(hydroxymethyl)-2-methylcyclopropyl) butyl-di-ynyl) benzoic acid (ACHL02-112·Intermediate-A)

試劑 MW 當量 Mmol g , mL 化合物11 268.31 1 1.5 400 mg LiOH H20 40.10 4 6 240 mg THF/H20(V/V=4:1) 20 mL 向攪拌之4-(((反）-2-(羥曱基）_2·曱基環丙基）丁-1，3-二炔基）苯甲酸曱酯11(400 mg，1.5 mm〇i；)於16 ml THF中之溶液中添加早水合風氧化裡（240 mg，6 mmol)於4 mL水中之溶液。添加後’混合物在室溫下持續擾拌15小時。當 LCMS顯示剩餘少量化合物11時，將混合物添加至50 ml水中。在0°C下用1 M HC1將pH值調節至3.0，用EtOAc(100 mLx3)萃取。經無水硫酸鈉乾燥經合併之有機層，濃縮得到370 mg呈黃色粉末狀之所需化合物，產率：92%。 LCMS: CP-0004344-006-3-03665-LCMSA035(ESI)m/z=255 (M+l)。4 NMR: CP-0004344-006-3 (DMSO-A，400 MHz) 6:13.1 (s, 1H), 7.91 (d, J=8.4 Hz, 2H), 7.62 (d, J=8.4 Hz, 2H), 4.75 (s, 1H), 3.22 (m, 2H), 1.59 (m, 1H), 1.18 (s, 3H), 1.08 (m, 1H)，0.66 (m, 1H) »Reagent MW equivalent Mmol g , mL Compound 11 268.31 1 1.5 400 mg LiOH H20 40.10 4 6 240 mg THF/H20 (V/V=4:1) 20 mL Stirring 4-(((trans)-2-(hydroxy) Addition of early hydration wind oxidation to a solution of decyl) 2 - decylcyclopropyl) butyl-1,3-diynyl benzoate 11 (400 mg, 1.5 mm 〇i;) in 16 ml of THF (240 mg, 6 mmol) in 4 mL of water. After the addition, the mixture was continuously spoiled for 15 hours at room temperature. When LCMS showed a small amount of compound 11 remaining, the mixture was added to 50 ml of water. The pH was adjusted to 3.0 with 1 M EtOAc over EtOAc (EtOAc (EtOAc) The combined organic layers were dried over anhydrous sodium sulfate and evaporated. LCMS: CP-0004344-006-3-03665-LCMS 035 (ESI) m/z = 255 (M+l). 4 NMR: CP-0004344-006-3 (DMSO-A, 400 MHz) 6:13.1 (s, 1H), 7.91 (d, J=8.4 Hz, 2H), 7.62 (d, J=8.4 Hz, 2H) , 4.75 (s, 1H), 3.22 (m, 2H), 1.59 (m, 1H), 1.18 (s, 3H), 1.08 (m, 1H), 0.66 (m, 1H) »

將（S)-2-胺基-3-(第三丁氧羰基胺基）-3-曱基丁酸甲酯 (347 mg，1.408 mmol，1.0 當量）及 4-(((lS，2R)-2-(羥曱基） 159849.doc -73- 201249786 2-曱基環丙基）丁 -1，3-二炔基）苯曱酸（358 mg，1.408 mmol’ 1.0當量）溶解於DMF(3 mL)中。向此添加DIPEA (0·615 mL，3·52 mmol，2.5 當量），接著添加 HATU(642 mg，1.689 mmol，1.2當量）。在室溫下攪拌此混合物約16 小時。使反應物於1 Μ檸檬酸與乙酸乙酯之間分配。用半飽和氯化鈉、飽和碳酸氫鈉，接著用飽和氯化鈉洗滌有機物，經硫酸鎂乾燥且蒸發至乾燥。得到912 mg粗（S)-3-(第三丁氧羰基胺基）-2-(4-(((lS，2R)-2-(羥甲基）·2-曱基環丙基）丁-1,3-二炔基）苯甲醯胺基）_3_甲基丁酸曱酯。LCMS Μ+1預期值=483.2，觀測值=483.2。Methyl (S)-2-amino-3-(t-butoxycarbonylamino)-3-mercaptobutanoate (347 mg, 1.408 mmol, 1.0 eq.) and 4-(((lS,2R)) -2-(Hydroxyfluorenyl) 159849.doc -73- 201249786 2-Mercaptocyclopropyl)butan-1,3-diynyl)benzoic acid (358 mg, 1.408 mmol' 1.0 eq.) was dissolved in DMF ( 3 mL). To this was added DIPEA (0·615 mL, 3.52 mmol, 2.5 eq.) followed by HATU (642 mg, 1.689 mmol, 1.2 eq.). The mixture was stirred at room temperature for about 16 hours. The reaction was partitioned between 1 EtOAc and EtOAc. The organics were washed with EtOAc (aq.) 912 mg of crude (S)-3-(t-butoxycarbonylamino)-2-(4-(((lS,2R)-2-(hydroxymethyl)) 2-indolylcyclopropyl) -1,3-Diynyl)benzhydrylamino)_3_methylbutyrate decyl ester. LCMS Μ+1 expected = 483.2, observed = 483.2.

將（S)-3-(第三丁氧羰基胺基）-2-(4-((( lS，2R)-2-(羥曱基）-2·曱基環丙基）丁-1,3-二炔基）苯甲醯胺基）-3-甲基丁酸曱酯 (粗物質）溶解於甲醇（1 mL)中。在室溫下添加含4.0 M HC1 之二 °惡烧（2.97 mL，11.86 mmol，8.43當量）。在60分鐘之後反應完成（HPLC)。在0°C下於旋轉蒸發器上濃縮反應物。向此添加IPA(2 mL)，接著添加羥胺溶液（1.859 mL， 28.1 mmol ’ 20當量）。將燒瓶置於4°C下約40小時。反應物濃縮（使反應物保持在0°C下）成膠黏物質。向此添加水（3 mL)及ACN(0.5 mL)。在0〇C下使用TFA(3 mL)酸化。再添加水（1 mL)及ACN(1 mL)» 藉由 RP HPLC(2"管柱，50 mL/min ’含0.1% TFA之水/ACN，在5% B下平衡）純化。使用注射過壚器（2x6.5 mL)加載於2"管柱（10 mL/min，5% B) 159849.doc •74· 201249786 上。經1分鐘勻變至50 mL/min。經66分鐘5%至30% B，在 25% B下產物溶離。合併所需溶離份，冷凍且置於冷凍乾燥器上。得到410 mg N-((S)-3·胺基-1-(羥胺基）·3_甲基側氧基丁 -2-基）-4-(((lS，2R)-2-(羥曱基）·2_甲基環丙基）丁 _ 1,3 - 一炔基）本曱酿胺（1-11) ’ TFA。LCMS Μ+1預期值 =384.2，觀測值=384.2。4 NMR (DMSO〇: δ 0.61-0.64 (t，1Η)，1.02-1.06 (m，1Η)，1.16 〇, 3Η), 1·25 (s，3Η), 1.30 (s, 3Η), 1.55-1.60 (m, 1H), 3.17-3.29 (m, 2H), 4.65-4.67 (d 1H), 7.61-7.63 (d, 2H), 7.88-7.90 (d, 2H)，7.98 (s，2H) 8.53-8.56 (d，1H)，9.21 (br，1H)，11.19 (s，1H)。 J·合成N-((S)-3-胺基-1-(經胺基）_3_甲基_1_側氧基丁基）-4-(((1,3-順）-3-(羥甲基）環丁基）丁·1>3_二炔基）苯甲酿胺（1-12)及N-((S)-3-胺基-1-(羥胺基）_3_甲基-1-側氧基丁·2_ 基）-4-(((1,3-反）-3-(經甲基）環丁基）丁 - i，3-二块基）苯甲酿胺（1_13)(S)-3-(Tertidinoxycarbonylamino)-2-(4-(((lS,2R)-2-(hydroxyindolyl)-2.nonylcyclopropyl)butane-1, 3-Diynyl)benzimidamide)-3-methylbutyrate decyl ester (crude) was dissolved in methanol (1 mL). A two-degree burn (4.07 mL, 11.86 mmol, 8.43 eq.) containing 4.0 M HCl was added at room temperature. The reaction was completed after 60 minutes (HPLC). The reaction was concentrated on a rotary evaporator at 0 °C. IPA (2 mL) was added thereto, followed by a hydroxylamine solution (1.859 mL, 28.1 mmol ' 20 eq.). The flask was placed at 4 ° C for about 40 hours. The reactants were concentrated (the reaction was maintained at 0 ° C) to form a sticky material. Water (3 mL) and ACN (0.5 mL) were added thereto. Acidified using TFA (3 mL) at 0 °C. Additional water (1 mL) and ACN (1 mL) were purified by RP HPLC (2 "column, 50 mL/min <0> Use a syringe (2 x 6.5 mL) to load on 2"column (10 mL/min, 5% B) 159849.doc •74·201249786. Change to 50 mL/min over 1 minute. The product was dissolved at 25% B over 5% to 30% B over 66 minutes. The desired fractions were combined, frozen and placed on a lyophilizer. Obtained 410 mg of N-((S)-3.amino-1-(hydroxylamino)·3_methyl-oxybutan-2-yl)-4-(((lS,2R)-2-(hydroxyl) Mercapto)·2_methylcyclopropyl)butyrate 1,1 - alkynyl) Benthoamine (1-11) 'TFA. LCMS Μ +1 expected value = 384.2, observed = 384.2. 4 NMR (DMSO 〇: δ 0.61-0.64 (t, 1 Η), 1.02-1.06 (m, 1 Η), 1.16 〇, 3 Η), 1·25 (s , 3Η), 1.30 (s, 3Η), 1.55-1.60 (m, 1H), 3.17-3.29 (m, 2H), 4.65-4.67 (d 1H), 7.61-7.63 (d, 2H), 7.88-7.90 ( d, 2H), 7.98 (s, 2H) 8.53-8.56 (d, 1H), 9.21 (br, 1H), 11.19 (s, 1H). J·Synthesis of N-((S)-3-amino-1-(amino)-3-(methyl-1-yloxybutyl)-4-(((1,3-cis)-3-) (Hydroxymethyl)cyclobutyl)butyl·1>3-diynyl)benzamide (1-12) and N-((S)-3-amino-1-(hydroxylamino)_3_A -1-yloxybutan-2-yl)-4-(((1,3-re)-3-(methyl)cyclobutyl)butan-i,3-diyl)benzamide (1_13)

ACHL02-146·順式 UBKi THF~ACHL02-146·cis UBKi THF~

TMPP, Pd(dba)4 TEA.DMF 159849.doc -75- 201249786 3-亞甲基環丁烷甲酸（2)TMPP, Pd(dba)4 TEA.DMF 159849.doc -75- 201249786 3-Methylenecyclobutanecarboxylic acid (2)

試劑 MW 當量 mol g，mL 化合物1 93 1.0 0.3 27.9 g KOH 56 5.0 1.5 84 g EtOH 200 mL H20 200 mL 3-亞曱基環丁烷甲腈1(27.9 g，0.3 mol)於乙醇（200 mL) 及水（200 mL)中之溶液用氫氧化钾（84 g，1.5 mol)處理。攪拌所得混合物且加熱至105°C維持4小時。減壓移除乙醇。使殘餘物冷卻至〇°C，且將pH值調節至1至2,接著用乙酸乙酯萃取，經硫酸鈉乾燥，過濾且移除溶劑。（30.2 g，產率=90%)。 3-亞甲基環丁烷甲酸甲酯（3>Reagent MW Equivalent mol g, mL Compound 1 93 1.0 0.3 27.9 g KOH 56 5.0 1.5 84 g EtOH 200 mL H20 200 mL 3-decylcyclobutanecarbonitrile 1 (27.9 g, 0.3 mol) in ethanol (200 mL) The solution in water (200 mL) was treated with potassium hydroxide (84 g, 1.5 mol). The resulting mixture was stirred and heated to 105 ° C for 4 hours. The ethanol was removed under reduced pressure. The residue was cooled to 〇 ° C, and the pH was adjusted to 1 to 2, then extracted with ethyl acetate, dried over sodium sulfate, filtered and solvent evaporated. (30.2 g, yield = 90%). Methyl 3-methylenecyclobutanecarboxylate (3>

試劑 MW 當量 mol g，mL 化合物2 112 1.0 0.260 29.12 g Me2S〇4 126 1.2 0.312 39.312 g K2CO3 138 2.0 0.520 70.2 g 丙嗣 500 mL 加熱3-亞甲基環丁烷甲酸2(29.12 g，0.26 mol)、碳酸鉀 (70.2 g，0·52 mol)、丙酮（500 mL)及硫酸二甲酯（39.312 g，0.3 12 mol)之混合物至回流，維持2小時。使反應混合物冷卻至室溫且過濾。減壓移除溶劑。用矽膠層析純化殘餘物，得到呈無色油狀之所需產物。（27.5 g，產率 = 84%)。 159849.doc -76- 201249786 3-(羥甲基）環丁烷甲酸甲酯（4) 試劑 MW 當量 mol g * mL 化合物3 126 1.0 0.210 26.46 g BH3 THF(1 M THF溶液） 0.3 0.070 70.0 mL THF 130 mL NaOH(3 Μ) 40 30 mL H2O2(30%) 34 1.0 0.210 34.0 g 乾燥三頸燒瓶裝入3-亞曱基環丁烷曱酸甲酯3(24.46 g， 0.21 mol)及無水THF(130 ml)，且冷卻至-10°C，接著經由注射器逐滴添加硼烷-THF錯合物（70.0 mL)。在室溫下攪拌所得混合物4小時，且冷卻至-20°C至-l〇°C ;添加甲醇，攪拌15分鐘。依序添加氫氧化鈉（3 Μ 30 mL)及過氧化氫 (3 4_0 g，0.210 mol)。攪拌混合物2小時且添加飽和亞硫酸鈉溶液（100 mL)。用水稀釋反應混合物，接著用乙酸乙酯萃取，用水及鹽水洗滌，經硫酸鈉乾燥，過濾，移除溶劑且用矽膠層析純化殘餘物。（28.73 g，95%)。 (反）-3-甲酿基環丁烷甲酸甲酯（5a)及（順）-3-甲醯基環丁烧甲酸甲酯（5b> 試劑 MW 當量 mmol g , mL 化合物4 144 1.0 69.4 l〇g 乙二醯二氣 128 2.0 139 17.8 g DMSO 78 4.0 278 21.7 g DCM 650 mL TEA 101 10.0 700 71 g 在-78°C下向乙二醯氯（17.8 g，139 mmol，2.0當量）於 CH2C12(45 0 mL)中之溶液中緩慢添加含DMSO(21.7 g，278 159849.doc -77- 201249786 mmol，4.0當量）之二氣甲烷（5〇 mL)。在3〇分鐘之後，在 -78°C下逐滴添加含3-(羥曱基）環丁烷甲酸曱酯4(10 g， 69.4 mmo卜1.0當量）之二氯曱烷（15〇 mL)。在_78。〇下再攪拌混合物2小時’且接著在_78°c下添加三乙胺（7〇 g，7〇〇 mmol ’ 10.0當量）。20分鐘之後，使混合物升溫至室溫，且添加飽和ΝΗβΙ水溶液。分離各層，且用二氣甲烷 (3x200 mL)萃取水層》用鹽水洗滌經合併之有機層，乾燥 (硫酸納）’過渡且濃縮，得到呈順式與反式異構體之混合物的粗產物。藉由二氧化矽管柱純化得到5a(反式，3〖g) 及 5b(順式，2.9 g)。 (順）-3-(2，2-二溴乙晞基）環丁烧甲酸甲酯（6b>Reagent MW equivalent mol g, mL Compound 2 112 1.0 0.260 29.12 g Me2S〇4 126 1.2 0.312 39.312 g K2CO3 138 2.0 0.520 70.2 g Propylene 500 mL Heated 3-methylenecyclobutanecarboxylic acid 2 (29.12 g, 0.26 mol) A mixture of potassium carbonate (70.2 g, 0. 52 mol), acetone (500 mL) and dimethyl sulfate (39.312 g, 0.312 mol) was refluxed for 2 hours. The reaction mixture was allowed to cool to room temperature and filtered. The solvent was removed under reduced pressure. The residue was purified by silica gel chromatography to give the desired product. (27.5 g, yield = 84%). 159849.doc -76- 201249786 Methyl 3-(hydroxymethyl)cyclobutanecarboxylate (4) Reagent MW Equivalent mol g * mL Compound 3 126 1.0 0.210 26.46 g BH3 THF (1 M in THF) 0.3 0.070 70.0 mL THF 130 mL NaOH (3 Μ) 40 30 mL H2O2 (30%) 34 1.0 0.210 34.0 g A dry three-necked flask was charged with methyl 3-mercaptocyclobutane decanoate 3 (24.46 g, 0.21 mol) and anhydrous THF ( 130 ml), and cooled to -10 ° C, then borane-THF complex (70.0 mL) was added dropwise via syringe. The resulting mixture was stirred at room temperature for 4 hours and cooled to -20 ° C to -10 ° C; methanol was added and stirred for 15 minutes. Sodium hydroxide (3 Μ 30 mL) and hydrogen peroxide (3 4_0 g, 0.210 mol) were added sequentially. The mixture was stirred for 2 hours and a saturated sodium sulfite solution (100 mL) was added. The reaction mixture was diluted with EtOAc (EtOAc)EtOAc. (28.73 g, 95%). Methyl (trans)-3-methylarylcyclobutanecarboxylate (5a) and methyl (cis)-3-carboxamidinecyclobutanecarboxylate (5b> reagent MW equivalent mmol g , mL compound 4 144 1.0 69.4 l 〇g 乙二醯二气128 2.0 139 17.8 g DMSO 78 4.0 278 21.7 g DCM 650 mL TEA 101 10.0 700 71 g to ethylenedichloride (17.8 g, 139 mmol, 2.0 eq.) at CH2C12 at -78 °C Dimethyl methane (5 〇 mL) containing DMSO (21.7 g, 278 159849.doc -77 - 201249786 mmol, 4.0 eq.) was slowly added to the solution in (45 0 mL). After 3 Torr, at -78 ° Add 3-(hydroxyxo)cyclobutanecarboxylate 4 (10 g, 69.4 mmo, 1.0 equivalent) of dichlorodecane (15 〇 mL) dropwise at C. Stir the mixture at _78. Triethylamine (7 〇g, 7 〇〇mmol '10.0 eq.) was added at _78 ° C. After 20 minutes, the mixture was allowed to warm to room temperature and a saturated aqueous solution of ΝΗβΙ was added. The layers were separated and The aqueous layer was extracted with di-methane (3 x 200 mL). The combined organic layers were washed with brine, dried (sodium sulfate) and then evaporated and concentrated to give a mixture of cis and trans isomers. The crude product was purified by ruthenium dioxide column to give 5a (trans, 3 〖g) and 5b (cis, 2.9 g). (cis)-3-(2,2-dibromoethyl) Cyclobutane methyl formate (6b>

試劑 MW 當量 mmol g，mL 化合物5a 142 1.0 20.40 2.9 g PPh3 262 4.0 81.80 21.41 g CBr4 DCM 332 2.0 40.90 13.56 g 200 mL 在低於-2(TC下在氬氣下向四溴化碳（13 56 g，40.89 mmol，2.0當量）於4〇 mL二氣曱烷令之溶液申逐滴添加三苯膦（21.41 g ’ 81.80 mmo卜4.0當量）於80 mL二氯甲烷中之溶液。混合物在-2〇t下持續攪拌30分鐘，接著冷卻至 -78 C »逐滴添加含（順）_3_甲醯基環丁烷甲酸甲酯5b(2 9 g ’ 2〇·4〇 mm〇h ! 〇當量）之8〇 mL二氯甲烷，且在_78〇c下再持續攪拌1小時。使混合物升溫至室溫。將反應混合物逐滴添加至攪拌之300 mL PE中。過濾混合物且用pe洗滌殘餘物。經硫酸鈉乾燥經合併之有機層，且濃縮得到目標 159849.doc •78· 201249786 化合物，藉由矽膠管柱純化。（2.3 g，產率=37%)。 ((順）-3-(2,2-二溴乙烯基）環丁基）甲醇（7b)Reagent MW equivalent mmol g, mL Compound 5a 142 1.0 20.40 2.9 g PPh3 262 4.0 81.80 21.41 g CBr4 DCM 332 2.0 40.90 13.56 g 200 mL At less than -2 (TC under argon to carbon tetrabromide (13 56 g) , 40.89 mmol, 2.0 eq.) A solution of triphenylphosphine (21.41 g '81.80 mmo 4.0 equivalent) in 80 mL of dichloromethane was added dropwise to a solution of 4 mL of dioxane. The mixture was at -2 Torr. Stirring was continued for 30 minutes at t, then cooled to -78 C » Add methyl (cis) _3_methyl decylcyclobutanecarboxylate 5b (2 9 g ' 2 〇 · 4 〇 mm 〇 h 〇 equivalent) 8 〇 mL of dichloromethane, and stirring was continued for an additional hour at _78 〇c. The mixture was allowed to warm to room temperature. The reaction mixture was added dropwise to a stirred 300 mL of PE. The mixture was filtered and washed with pe. The combined organic layers were dried <RTI ID=0.0>(~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ 2,2-dibromovinyl)cyclobutyl)methanol (7b)

試劑 MW 當量 mmol g > mL 化合物6b 298 1.0 8.4 2.5 g L1BH4 22 1.1 9.2 203 mg THF 10 mL 在〇°C下向化合物6b(2.5 g，8.4 mmol，1.0當量）溶解於 THF(1 0 mL)中之溶液中添加硼氫化裡（203 mg)。反應混合物在室溫下持續攪拌3小時，添加水（5 mL)且用乙酸乙酯萃取。乾燥經合併之乙酸乙酯層且減壓濃縮。藉由矽膠管柱用PE:EA=5:1純化粗產物。（1.88 g，產率=83%)。 4-(((順>-3-(羥甲基> 環丁基）丁-1，3-二炔基> 苯甲酸甲酯（9b) 試劑 MW 當量 mmol g , mL 化合物7b 270 1.0 6.9 1-88 g 化合物8 160 1.4 9.7 1.56 g TMPP 352 0.04 0.28 98 mg Pd(dba)4 916 0.01 0.069 64 mg DMF 30 mL TEA 101 3.0 20.9 2.11 g 在氮氣氛圍下向（（順）-3-(2,2-二溴乙烯基）環丁基）甲醇 7b(1.88 g，6.9 mmol，1.0當量）、4-乙炔基苯甲酸甲酯 8(1.56 g，9.7 mmol，1.4 當量）、三（4-甲基苯基）膦（98 mg，0.28 mmol，0.04 當量）、Pd(dba)4(64 mg，0.069 mmol，0.01當量）於無水DMF(30 mL)中之混合物中添加三乙胺（2.11 g，20.9 mmol，3.0當量）。反應混合物在80°C下持續攪拌15小時。LCMS監測反應。接著用NH4C1水溶液 159849.doc -79- 201249786 稀釋反應混合物，且用乙酸乙酯萃取。用水、鹽水洗滌經合併之乙酸乙酯層且乾燥。減壓蒸發溶劑得到粗物質，藉由石夕膠管柱用PE:EA=5 :1純化。（450 mg，產率=24%)。 4-(((順）-3-(羥甲基）環了基）丁 -1，3-二炔基）苯甲酸 (ACHL02-146-順式)Reagent MW Equivalent mmol g > mL Compound 6b 298 1.0 8.4 2.5 g L1BH4 22 1.1 9.2 203 mg THF 10 mL Compound 6b (2.5 g, 8.4 mmol, 1.0 eq.) was dissolved in THF (10 mL) at EtOAc. Boron hydride (203 mg) was added to the solution. The reaction mixture was stirred at room temperature for 3 hrs, water (5 mL) andEtOAc. The combined ethyl acetate layers were dried and concentrated under reduced pressure. The crude product was purified by a silica gel column using PE:EA = 5:1. (1.88 g, yield = 83%). 4-(((cis >-3-(hydroxymethyl> cyclobutyl)butan-1,3-diynyl> methyl benzoate (9b) reagent MW equivalent mmol g , mL Compound 7b 270 1.0 6.9 1-88 g Compound 8 160 1.4 9.7 1.56 g TMPP 352 0.04 0.28 98 mg Pd(dba)4 916 0.01 0.069 64 mg DMF 30 mL TEA 101 3.0 20.9 2.11 g Under nitrogen atmosphere ((cis)-3-( 2,2-Dibromovinyl)cyclobutyl)methanol 7b (1.88 g, 6.9 mmol, 1.0 eq.), methyl 4-ethynylbenzoate 8 (1.56 g, 9.7 mmol, 1.4 eq.), Methylphenyl)phosphine (98 mg, 0.28 mmol, 0.04 eq.), Pd (dba) 4 (64 mg, 0.069 mmol, 0.01 eq.) 20.9 mmol, 3.0 eq.) The reaction mixture was stirred for 15 h at 80 ° C. The reaction was monitored by LCMS. The reaction mixture was then diluted with aqueous NH4C1 159849.doc -79 - 201249786 and extracted with ethyl acetate. The combined ethyl acetate layers were dried and evaporated <RTI ID=0.0></RTI>jjjjjjjjjjjjjjjjjjj (cis)-3-(hydroxymethyl)cyclo)butyl-1,3-diynyl)benzoic acid (ACHL02-146-cis)

試劑 MW 當量 mmol g，mL 化合物9a 268 1.0 1.68 450 mg LiOH H20 42 4.0 6.72 282 mg THF 20 mL 向4-(((順）-3-(羥甲基）環丁基）丁-1，3-二炔基）苯曱酸甲酯 9b(450 mg，1.0當量）於THF(200 mL)中之溶液中添加單水合氫氧化鋰（282 mg，4.0當量），接著混合物在25°C下反應 5小時。接著移除反應溶劑且用3 N HC1中和至pH值5至6，用乙酸乙酯萃取。用鹽水洗滌經合併之乙酸乙酯層且乾燥。蒸發溶劑獲得目標化合物。（400 mg，產率=94%)。 (反）-3-(2,2-二溴乙烯基）環丁烷甲酸甲酯（6a)Reagent MW equivalent mmol g, mL Compound 9a 268 1.0 1.68 450 mg LiOH H20 42 4.0 6.72 282 mg THF 20 mL to 4-(((cis)-3-(hydroxymethyl)cyclobutyl)butane-1,3- Add a solution of methyl diacetyl)benzoate (9 mg (450 mg, 1.0 eq.) in THF (200 mL). hour. The reaction solvent was then removed and neutralized with 3 N HCl to pH 5 to 6 and extracted with ethyl acetate. The combined ethyl acetate layers were washed with brine and dried. Evaporation of the solvent gave the title compound. (400 mg, yield = 94%). (trans)methyl-3-(2,2-dibromovinyl)cyclobutanecarboxylate (6a)

試劑 MW 當量 Mmol g，mL 化合物5a 142 1.0 16.90 2.4 g PPh3 262 4.0 67.60 17.7 g CBr4 332 2.0 33.80 11.2g DCM 200 mL 在低於-20°C下在氬氣下向四溴化碳（11.2 g，33.80 mmol，2.0當量）於40 mL二氯甲烧中之溶液中逐滴添加三苯膦（17.7 g，67.60 mmol，4.0當量）於80 mL二氯甲烷中之溶液。混合物在-20°C下持續攪拌30分鐘，接著冷卻至 159849.doc -80- 201249786 -78 C。逐滴添加含（反）-3-甲醯基環丁院曱酸曱酯5a(2.4 g ’ 16.90 mmoh lo 當量）之 8〇 mL 二氣曱烷，且在 _78〇ct 再持續攪拌1小時。使混合物升溫至室溫，逐滴添加反應混合物至攪拌之300 mL PE中。過濾且用pE洗滌殘餘物。經硫酸鈉乾燥經合併之有機層且濃縮得到目標化合物，藉由碎膠管柱純化。（2.5 g，產率=50%)。 ((反）-3-(2,2-二溴乙烯基）環丁基}甲醇（7a)Reagent MW Equivalent Mmol g, mL Compound 5a 142 1.0 16.90 2.4 g PPh3 262 4.0 67.60 17.7 g CBr4 332 2.0 33.80 11.2g DCM 200 mL Under argon to argon carbon tetrabromide (11.2 g, below -20 ° C, A solution of triphenylphosphine (17.7 g, 67.60 mmol, 4.0 eq.) in 80 mL of dichloromethane was added dropwise to a solution of <RTIgt; The mixture was continuously stirred at -20 ° C for 30 minutes and then cooled to 159849.doc -80 - 201249786 -78 C. 8 〇 mL of dioxane containing (trans)-3-carboxamidine butyl decanoate 5a (2.4 g ' 16.90 mmoh lo equivalent) was added dropwise, and stirring was continued for 1 hour at _78 〇ct. . The mixture was allowed to warm to room temperature and the reaction mixture was added dropwise to a stirred 300 mL PE. Filter and wash the residue with pE. The combined organic layers were dried over sodium sulfate and concentrated to give title compound (2.5 g, yield = 50%). ((trans)-3-(2,2-dibromovinyl)cyclobutyl}methanol (7a)

試劑 MW 當量 mmol g，mL 化合物6a 298 1.0 8.4 2.5 g L1BH4 THF 22 1.1 9.2 203 mg 10 mL 在〇°C下向（反）-3-(2,2-二溴乙烯基）環丁烷曱酸甲酯 6a(2_5 g，8.4 mmol，1.0當量）溶解於THF(10 mL)中之溶液中添加硼氫化鋰（203 mg)。反應混合物在室溫下持續授拌 3小時’添加水（5 mL)且用乙酸乙酯萃取。乾燥經合併之乙酸乙酯層且減壓濃縮。藉由矽膠管柱用pe:Ea=5:丨純化粗物質。（1.70 g，產率=75%)。 4-(((反）-3·(羥甲基）環丁基）丁-13-二炔基>苯f酸f ^(9α) 試劑 MW 當量 mmol g，mL 化合物7a 270 1.0 6.3 1.70 g 化合物8 160 1.4 8.8 1.41 g TMPP 352 0.04 0.25 89 mg Pd(dba)4 916 0.01 0.063 58 mg DMF 30 mL TEA 101 3.0 18.9 1.91 ε 在氮氣下向（（反）-3-(2,2-二溴乙烯基）環丁基）甲醇7a 159849.doc -81 · 201249786 (1.70 g ’ 6.3 mmol ’ 1.0 當量）、化合物 8(1.41 g，8.8 mmol，1·4當量）、三（4-甲基苯基）膦（89 mg，0.25 mm〇l， 0.04當量）、Pd(dba)4(58 mg，0.063 mmol，0.01 當量）於無水DMF(30 mL)中之混合物中添加三乙胺g 9i g，18 9 mmol，3.0當量）*反應混合物在80°C下持續授拌15小時。 LCMS監測反應。用ΝΗβΙ水溶液稀釋反應混合物，且用乙酸乙酯萃取。用水、鹽水洗滌經合併之乙酸乙醋層且乾燥。減壓蒸發溶劑得到粗物質，藉由石夕膠管枉用 PE:EA=5:1 純化。（550 mg，產率=33%)。 4-(((反）-3-(經甲基）環丁基）丁 -i，3-二炔基）苯甲酸 (ACHL02-146-反式)Reagent MW equivalent mmol g, mL Compound 6a 298 1.0 8.4 2.5 g L1BH4 THF 22 1.1 9.2 203 mg 10 mL (trans)-3-(2,2-dibromovinyl)cyclobutane decanoic acid at 〇 ° C Methylborohydride (203 mg) was added to a solution of methyl ester 6a (2_5 g, 8.4 mmol, 1.0 eq.) in THF (10 mL). The reaction mixture was continuously stirred at room temperature for 3 hours. Water (5 mL) was added and extracted with ethyl acetate. The combined ethyl acetate layers were dried and concentrated under reduced pressure. The crude material was purified by a ruthenium tube column with pe: Ea = 5: hydrazine. (1.70 g, yield = 75%). 4-(((trans)-3·(hydroxymethyl)cyclobutyl)but-13-diynyl> benzene f acid f^(9α) Reagent MW equivalent mmol g, mL Compound 7a 270 1.0 6.3 1.70 g Compound 8 160 1.4 8.8 1.41 g TMPP 352 0.04 0.25 89 mg Pd(dba)4 916 0.01 0.063 58 mg DMF 30 mL TEA 101 3.0 18.9 1.91 ε Under nitrogen ((trans)-3-(2,2-dibromo) Vinyl)cyclobutyl)methanol 7a 159849.doc -81 · 201249786 (1.70 g '6.3 mmol '1.0 equivalent), compound 8 (1.41 g, 8.8 mmol, 1.4 equivalent), tris(4-methylphenyl) Add a mixture of phosphine (89 mg, 0.25 mm ,l, 0.04 eq.), Pd (dba) 4 (58 mg, 0.063 mmol, 0.01 eq.) in anhydrous DMF (30 mL). 9 mmol, 3.0 eq.) * The reaction mixture was continuously stirred at 80 ° C for 15 hours. The reaction was monitored by LCMS. The reaction mixture was diluted with ΝΗβΙ aqueous solution and extracted with ethyl acetate. The combined ethyl acetate layer was washed with water and brine and dried. The solvent was evaporated under reduced pressure to give a crude material. m. (550 mg, yield = 33%). 4-(((trans)-3-(methyl)cyclobutyl)butan-i,3-diynyl)benzoic acid (ACHL02-146-trans)

試劑 MW 當量 mmol g , mL 化合物9a 268 1.0 2.05 550 mg LiOH H20 42 4.0 8.21 345 mg THF 20 mL 向4-(((反）-3-(羥甲基）環丁基）丁-1，3-二炔基）苯甲酸甲酯 9a(550 mg，1.0當量）於THF(20 mL)中之溶液中添加單水合氫氧化鋰（345 mg ’ 4.0當量）’接著混合物在25。(：下反應5 小時。接著移除反應溶劑且用3 N HC1中和至PH值5至6，用乙酸乙醋萃取。用鹽水洗蘇經合併之乙酸乙酯層且乾燥。蒸發溶劑獲得目標化合物。（340 mg，產率=65°/〇)。Reagent MW equivalent mmol g , mL Compound 9a 268 1.0 2.05 550 mg LiOH H20 42 4.0 8.21 345 mg THF 20 mL to 4-(((trans)-3-(hydroxymethyl)cyclobutyl)butane-1,3- To a solution of methyl diacetyl)benzoate 9a (550 mg, 1.0 eq.) in THF (20 mL) EtOAc (EtOAc &lt (The reaction was carried out for 5 hours. The reaction solvent was then removed and neutralized with 3 N HCl to pH 5 to 6 and extracted with ethyl acetate. The combined ethyl acetate layer was washed with brine and dried. Compound (340 mg, yield = 65 ° / 〇).

159849.doc -82 ·159849.doc -82 ·

S 201249786 將（S)-2-胺基-3-(第三丁氧羰基胺基）_3_曱基丁酸曱酯 (387 mg，1.573 mmol，1.0 當量）及 4-(((1，3-順）-3-(羥甲基）環丁基）丁-1，3-二炔基）苯甲酸（4〇〇 mg，1.573 mmol，1.0 當量）溶解於DMF(3 mL)中。向此添加DIPEA(0.687 mL， 3.93 mmol，2.5 當量），接著添加 HATU(718 mg，1.888 mmol，1.2當量）^在室溫下攪拌此混合物約16小時。使反應物於1 Μ檸檬酸與乙酸乙酯之間分配。用半飽和氣化納、飽和破酸氫鈉，接著用飽和氯化鈉洗滌有機物，經硫酸鎂乾燥且蒸發至乾燥。得到1〇35 g粗（s)_3_(第三丁氧羰基胺基）-2-(4-(((1,3-順）-3-(羥曱基）環丁基）丁 _i，3-二炔基）苯曱酿胺基）-3-甲基丁酸甲酯。LCms M+1預期值= 483.2，觀測值=483.2。S 201249786 (S)-2-Amino-3-(t-butoxycarbonylamino)-3-mercaptobutyrate (387 mg, 1.573 mmol, 1.0 eq.) and 4-((1,3) -cis-3-(Hydroxymethyl)cyclobutyl)butan-1,3-diynyl)benzoic acid (4 mg, 1.537 mmol, 1.0 eq.) was dissolved in DMF (3 mL). To this was added DIPEA (0.687 mL, 3.93 mmol, 2.5 eq.), followed by HATU (718 mg, 1.88 mmol, 1.2 eq.). The mixture was stirred at room temperature for about 16 hours. The reaction was partitioned between 1 citric acid and ethyl acetate. The organics were washed with a saturated solution of sodium sulphate and then washed with saturated sodium chloride, dried over magnesium sulfate and evaporated to dry. Obtaining 1〇35 g of crude (s)_3_(t-butoxycarbonylamino)-2-(4-((1,3-(cis)-3-(hydroxy)yl)cyclobutyl)- 3-Diynyl) Benzoquinone Aminomethyl 3-methylbutanoate. LCms M+1 expected = 483.2, observed = 483.2.

將（S)-3_(第三丁氧羰基胺基）_2·(4_(((1，3順）_3_(羥甲基）環丁基）丁 -1,3-二炔基）苯甲醯胺基）_3_曱基丁酸甲酯（粗物質）溶解於曱醇（1 mL)中。在室溫下添加含4 〇 μ HC1之二噁烷（3.3 mL，13.26 mmol，8.43當量）。在60分鐘之後反應完成（HPLC)。在〇。〇下於旋轉蒸發器上濃縮反應物。向此添加IPA(2 mL) ’接者添加經胺溶液（2 mL，3 1.5 mmol， 20當量）。將燒瓶置於4〇Ct24小時。反應物濃縮（使反應物保持在〇c下）成膠黏物質，向此添加水（3 mL)及ACN(0.5 mL)。在（TC下使用TFA(3 mL)酸化。再添加水（i mL)及 159849.doc • 83 - 201249786 ACN(1 mL)。藉由 RP HPLC(2"管柱，50 mL/min，含〇.1% TFA之水/ACN ’在5 % B下平衡）純化。使用注射過據器 (2x6.5 mL)加載於2"管柱（10 mL/min，5% 6)上》經1分鐘勻變至50 1^/11^11。經55分鐘5%至35%8，在24.5〇/〇至26〇/〇 B下產物溶離。合併所需溶離份，冷凍且置於冷凍乾燥器上。得到460 mg N-((S)-3-胺基-1-(羥胺基）-3-曱基_1_側氧基丁-2-基）-4-(((1，3-順）-3-(羥甲基）環丁基）丁-1，3_二炔基）苯甲醯胺’ TFA。LCMS M+1預期值=384.2，觀測值 =384.2。NMR (DMSO-A): δ 1.25 (s，3H)，1.30 (s，3H)， 1.81-1.88 (m, 2H), 2.23-2.34 (m, 3H), 3.08-3.15 (m, 1H), 3.30-3.31 (d, 1H), 4.54 (br, 1H), 4.65-4.67 (d, 1H), 7.61-7.64 (d, 2H), 7.89-7.91 (d, 2H), 7.99 (s, 2H), 8.54-8.56 (d, 1H), 9.22 (br，1H)，11.20 (s, 1H)。(S)-3_(Tertidinoxycarbonylamino)_2·(4_((1,3 cis)_3_(hydroxymethyl)cyclobutyl)butan-1,3-diynyl)benzimidazole Methylamino)_3_mercaptobutyrate (crude) was dissolved in methanol (1 mL). Dioxane (3.3 mL, 13.26 mmol, 8.43 eq.) containing 4 〇 μ HCl was added at room temperature. The reaction was completed after 60 minutes (HPLC). Here. The reaction was concentrated on a rotary evaporator. To this was added IPA (2 mL). The amine solution (2 mL, 3 1.5 mmol, 20 eq.) was added. The flask was placed at 4 ° Ct for 24 hours. The reactants were concentrated (the reaction was kept under 〇c) to form a gummy material, and water (3 mL) and ACN (0.5 mL) were added. Acidified with TFA (3 mL) under TC. Add water (i mL) and 159849.doc • 83 - 201249786 ACN (1 mL) by RP HPLC (2 "column, 50 mL/min, containing hydrazine .1% TFA water/ACN 'equalized at 5% B.) Purified using a syringe (2 x 6.5 mL) loaded onto a 2"column (10 mL/min, 5% 6) for 1 minute Rough to 50 1^/11^11. Dissolve the product at 24.5 〇/〇 to 26 〇/〇B over 55 minutes 5% to 35% 8. Combine the desired fractions, freeze and place on a freeze dryer 460 mg of N-((S)-3-amino-1-(hydroxyamino)-3-indolyl-1-yloxybut-2-yl)-4-(((1,3-(cis) -3-(hydroxymethyl)cyclobutyl)butane-1,3-diynyl) benzalkonamine 'TFA. LCMS M+1 expected = 384.2, observed = 384.2. NMR (DMSO-A) : δ 1.25 (s, 3H), 1.30 (s, 3H), 1.81-1.88 (m, 2H), 2.23-2.34 (m, 3H), 3.08-3.15 (m, 1H), 3.30-3.31 (d, 1H ), 4.54 (br, 1H), 4.65-4.67 (d, 1H), 7.61-7.64 (d, 2H), 7.89-7.91 (d, 2H), 7.99 (s, 2H), 8.54-8.56 (d, 1H ), 9.22 (br, 1H), 11.20 (s, 1H).

159849.doc • 〇4 - S159849.doc • 〇4 - S

將（S)-2-胺基-3-(第三丁氧羰基胺基）·3_甲基丁酸曱酯 (329 mg’ 1.377 mmol，1.0 當量）及 4-(((1,3-反）-3-(羥曱基）環丁基）丁-1，3-二快基）苯甲酸（3 40 mg，1.377 mmol，1.0 當量）溶解於DMF(3 mL)中。向此添加DIPEA(0.584 mL， 3.34 mmol ’ 2.5 當量）’接著添加 HATU(610 mg，1.605 mmol ’ 1.2當量）。在室溫下攪拌此混合物約16小時。使反應物於1 Μ檸檬酸與乙酸乙酯之間分配。用半飽和氣化納、飽和碳酸氫鈉’接著用飽和氯化鈉洗滌有機物，經硫酸鎂乾燥且蒸發至乾燥。得到990 mg粗（S)-3-(第三丁氧羰 201249786 基胺基）-2-(4-(((1,3-反）-3-(羥曱基）環丁基）丁-1，3-二炔基）苯甲醯胺基）-3-甲基丁酸甲酯》LCMS M+1預期值=483.2，觀測值=483.2。(S)-2-Amino-3-(t-butoxycarbonylamino)-3-methylbutyrate (329 mg ' 1.377 mmol, 1.0 eq.) and 4-(((( </RTI> </RTI> <RTIgt; </RTI> <RTIgt; </RTI> <RTIgt; </RTI> <RTIgt; </RTI> <RTIgt; To this was added DIPEA (0.584 mL, 3.34 mmol '2.5 eq.). then HATU (610 mg, 1.605 mmol. The mixture was stirred at room temperature for about 16 hours. The reaction was partitioned between 1 citric acid and ethyl acetate. The organics were washed with a saturated solution of sodium sulphate and then saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness. 990 mg of crude (S)-3-(t-butoxycarbonyl 201249786 amino)-2-(4-((1,3-))-3-(hydroxyindenyl)cyclobutyl)- Methyl 1,3-diynyl) benzhydrazinyl)-3-methylbutanoate. LCMS M+1 expected = 483.2, observed = 483.2.

將（S)-3-(第三丁氧羰基胺基）-2-(4-(((1,3-反）-3-(羥曱基）環丁基）丁 -1,3-二炔基）苯甲醯胺基）-3 -甲基丁酸曱酯（粗物質）溶解於曱醇（1 mL)中。在室溫下添加含4.0 M HC1之二噁烧（2.82 mL ’ 11.27 mmol，8.43當量）。在60分鐘之後反應完成（HPLC)。在0°C下於旋轉蒸發器上濃縮反應物。向此添加IPA(2 mL) ’接著添加羥胺溶液（1.77 mL，26.7 mmol，20當量）。將燒瓶置於4°C下96小時。接著將反應物濃縮（使反應物保持在0°C下）成膠黏物質。向此添加水（3 mL)及ACN(0.5 mL)。在〇°C下使用TFA(3 mL)酸化溶液。再添加水（1 mL)及ACN(1 mL)。藉由RP HPLC(2"管柱，50 mL/min，含〇·1% TFA之水/ACN，在5% B下平衡）純化化合物；使用注射過濾器（2x7 mL)加載於2"管柱（1〇 mL/min， 5%B)上；經1分鐘勻變至5〇 mL/min且經55分鐘5%至35〇/〇 B。在22_9°/。至25·5% B下產物溶離。合併所需溶離份，冷凍且置於冷凍乾燥器上。該製程得到41〇 mg 基-1-(羥胺基）-3-甲基-1-側氧基丁 _2_基反•(羥曱基）環丁基）丁 -1，3-二炔基）笨曱醢胺，tfa。LCMS 159849.doc 201249786 M+l，預期值=384.2 ’ 觀測值=384.2。1HNMR(DMSO-d6): δ 1.25 (s, 3H), 1.30 (s, 3H), 2.07-2.11 (t, 4H), 2.40-2.43 (m, 1H), 3.22-3.26 (m, 1H), 3.37-3.38 (d, 1H), 4.57 (br, 1H), 4.65-4.67 (d, 1H), 7.62-7.64 (d, 2H), 7.89-7.91 (d, 2H), 7.97 (s, 2H), 8.54-8.56 (d, 1H), 9.21 (br, 1H), 11.19 (s，1H) 〇 K.合成N-((S)-3-胺基-1-(羥胺基）-3-甲基-1-側氧基丁-2-基）-4-(((1,3-反）-3-(羥甲基）環戊基）丁-1，3-二炔基）苯甲酿胺（1-14)(S)-3-(Tertiary butoxycarbonylamino)-2-(4-(((1,3-))-3-(hydroxy)]cyclobutyl)butane-1,3-di The alkynyl)benzimidamide-3-methylbutyrate (crude) was dissolved in decyl alcohol (1 mL). Dioxoaldehyde (2.82 mL ' 11.27 mmol, 8.43 eq.) containing 4.0 M HCl was added at room temperature. The reaction was completed after 60 minutes (HPLC). The reaction was concentrated on a rotary evaporator at 0 °C. To this was added IPA (2 mL), followed by a solution of hydroxylamine (1.77 mL, 26.7 mmol, 20 eq.). The flask was placed at 4 ° C for 96 hours. The reaction was then concentrated (the reaction was maintained at 0 ° C) to form a gum. Water (3 mL) and ACN (0.5 mL) were added thereto. The solution was acidified using TFA (3 mL) at 〇 °C. Additional water (1 mL) and ACN (1 mL) were added. The compound was purified by RP HPLC (2 "column, 50 mL/min, water containing 〇·1% TFA/ACN, equilibrated at 5% B); loaded with 2"column using a syringe filter (2 x 7 mL) (1 〇 mL / min, 5% B); ramped to 5 〇 mL / min over 1 minute and 5% to 35 〇 / 〇 B over 55 minutes. At 22_9°/. The product was dissolved to 25.5% B. The desired fractions were combined, frozen and placed on a freeze dryer. The process yielded 41 mg of 1-(hydroxylamino)-3-methyl-1-oxobutan-2-yl-(hydroxyxanyl)cyclobutyl)butane-1,3-diynyl ) clumamine, tfa. LCMS 159849.doc 201249786 M+l, expected value = 384.2 ' observed value = 384.2. 1H NMR (DMSO-d6): δ 1.25 (s, 3H), 1.30 (s, 3H), 2.07-2.11 (t, 4H), 2.40-2.43 (m, 1H), 3.22-3.26 (m, 1H), 3.37-3.38 (d, 1H), 4.57 (br, 1H), 4.65-4.67 (d, 1H), 7.62-7.64 (d, 2H ), 7.89-7.91 (d, 2H), 7.97 (s, 2H), 8.54-8.56 (d, 1H), 9.21 (br, 1H), 11.19 (s, 1H) 〇K. Synthesis N-((S) 3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)-4-(((1,3-re)-3-(hydroxymethyl)) ring Pentyl)butyl-1,3-diynyl)benzamide (1-14)

ACHL02-149 (順）-環戊烷-hk二甲酸(2)ACHL02-149 (cis)-cyclopentane-hk dicarboxylic acid (2)

試劑 MW 當量 mmol g，mL 化合物1 94 1.0 106 10 g ΚΜη04 158 2.8 297 47 g MgS〇4 120 0.33 35.4 4.3 g h2o 500 mL 丙_ 15 mL 向冷卻至5°C之攪拌之高錳酸鉀（47 g，0.297 mol)及硫酸錯（4.3 g，35.4 mmol)於水（500 mL)中之懸浮液中整份添加雙環[2.2.1]庚 _2-烯 1(10 g，0.106 mol)於丙酮（15 mL)中之溶液（5°C至40°C)。移除冰浴且再攪拌反應物2小時。此 159849.doc -86 - 201249786 後，過濾混合物且用亞硫酸氫鈉處理濾液且用1 N HC1酸化至pH=2。接著用乙酸乙酯（800 mLx3)萃取且用水（1 L)、鹽水（1 L)洗滌經合併之有機層，乾燥，濃縮得到呈白色固體狀之目標化合物2(7.5 g，46%)，其未經純化即用於下一步驟。 (順）-環戊烷-13-二甲酸二甲酯（3)Reagent MW equivalent mmol g, mL Compound 1 94 1.0 106 10 g ΚΜη04 158 2.8 297 47 g MgS〇4 120 0.33 35.4 4.3 g h2o 500 mL Propion _ 15 mL Stirred potassium permanganate cooled to 5 ° C (47 g, 0.297 mol) and sulfuric acid (4.3 g, 35.4 mmol) in water (500 mL) in a suspension of bicyclo [2.2.1] hept-2-ene 1 (10 g, 0.106 mol) in acetone Solution in (15 mL) (5 ° C to 40 ° C). The ice bath was removed and the reaction was stirred for additional 2 hours. After this 159849.doc -86 - 201249786, the mixture was filtered and the filtrate was treated with sodium bisulfite and acidified to pH = 2 with 1 N HCl. The title compound 2 (7.5 g, 46%) was obtained as a white solid, which was taken from ethyl acetate (yield: EtOAc, EtOAc) It was used in the next step without purification. (cis)-cyclopentane-13-dicarboxylic acid dimethyl ester (3)

試劑 MW 當量 mmol g，mL 化合物2 158 1.0 47.5 7.5 g S0C12 119 4.0 190 22.6 g MeOH 200 mL 在冰浴下向攪拌之（順）-環戊烷-1,3-二曱酸2(7.5 g，47.5 mmol)於甲醇（200 mL)中之溶液中添力口亞硫醯氣（22_6 g， 0.19 mol)。接著加熱反應混合物至回流隔夜。此後，濃縮混合物得到呈黃色油狀之化合物3(8.3 g，94%)，其未經純化用於下一步驟。藉由GC-MS確認。環戍烷-1，3-二基二f醇（4) 試劑 MW 當量 mmol g，mL 化合物2 186 1 44.6 8.3 g LiAlH4 38 2.0 89.2 3.4 g THF 150 ml 在冰浴下向攪拌之LAH(3.4 g，89.2 mmol)於THF(100 mL)中之溶液中添加（順）-環戊烷-1，3-二甲酸二甲酯3(8_3 g，44.6 mmol)於THF(50 mL)中之溶液。接著在室溫下擾拌反應混合物隔夜。此後，用水（6.5 mL)稀釋混合物，接著過濾且濃縮濾液，用矽膠層析（PE:EA=2:1)純化殘餘物 159849.doc • 87 · 201249786 得到呈黃色油狀之化合物4(4.0 g，69%)。H NMR (400 MHz, DMSO-i/d) 0.53-0.50 (m, 1H), 0.96-1.00 (m, 2H), 1.29-1.35 (m, 2H), 1.47-1.54 (m, 1H), 1.66-1.73 (m, 2H), 2.98-3.01 (m，4H)，4.29 (s，1H)。乙酸3-(羥甲基）環戊基）甲酯（5)Reagent MW equivalent mmol g, mL Compound 2 158 1.0 47.5 7.5 g S0C12 119 4.0 190 22.6 g MeOH 200 mL Stirring (cis)-cyclopentane-1,3-didecanoic acid 2 (7.5 g, in an ice bath) 47.5 mmol) of sulfite gas (22_6 g, 0.19 mol) was added to a solution of methanol (200 mL). The reaction mixture was then heated to reflux overnight. After this time the mixture was concentrated to give compound 3 (8.3 g, 94% Confirmed by GC-MS. Cyclodecane-1,3-diyldi-fol (4) Reagent MW Equivalent mmol g, mL Compound 2 186 1 44.6 8.3 g LiAlH4 38 2.0 89.2 3.4 g THF 150 ml LAH (3.4 g) stirred under ice bath A solution of (cis)-cyclopentane-1,3-dicarboxylic acid dimethyl ester 3 (8-3 g, 44.6 mmol) in THF (50 mL). The reaction mixture was then stirred overnight at room temperature. After that, the mixture was diluted with water (6.5 mL), then filtered, and the filtrate was evaporated, mjjjjjjjjjjjjjjjjjjjj , 69%). H NMR (400 MHz, DMSO-i/d) 0.53-0.50 (m, 1H), 0.96-1.00 (m, 2H), 1.29-1.35 (m, 2H), 1.47-1.54 (m, 1H), 1.66- 1.73 (m, 2H), 2.98-3.01 (m, 4H), 4.29 (s, 1H). 3-(hydroxymethyl)cyclopentyl)methyl acetate (5)

試劑 MW 當量 mmol g，mL 化合物4 130 1.0 31 4.0 g AcCl 78 1.0 34 2.7 g NaH(60°/〇) 24 0.9 28 1.1 g THF 60 mL 在冰浴下向攪拌之環戊烷-1,3-二基二甲醇4(4.0 g，31 mmol)於THF(60 mL)中之溶液中添加NaH(60%)(l.l g，28 mol)。接著在冰浴下攪拌反應混合物0.5小時，接著將乙醯氯（2.7 g，34 mmol)添加至反應混合物中。且在室溫下攪拌混合物隔夜。此後，用水（1.0 mL)稀釋混合物，接著過濾且濃縮濾液，用矽膠層析純化殘餘物得到呈黃色油狀之化合物 5(1.7 g，25%)。NMR (400 MHz，DMSO-A) 0.84-0.87 (m, 1H), 1.28-1.31 (m, 2H), 1.62-1.65 (m, 2H), 1.80-1.83 (m, 1H), 1.99 (s, 1H), 2.10-2.18 (m, 1H), 3.26-3.29 (m，2H)，3.89 (d，·7=8, 2H)，4.43 (s，1H)。乙酸（3-甲醮基環戊基）f酯（6) 試劑 MW 當量 mmol g , mL 化合物5 172 1 9.9 1-7 g PCC 215 3.0 29.7 6.4 g CH2C12 50 ml -88· 159849.doc s 201249786 向攪拌之乙酸（3-(羥曱基）環戊基）甲酯5(1.7 g，9.9 mmol)於CH2Cl2(50 mL)中之溶液中逐份添加PCC(6.4 g， 29.7 mmol)。接著在室溫下攪拌反應混合物隔夜。此後，濃縮混合物，用矽膠層析（PE:EA=6:1)純化殘餘物得到呈黃色油狀之化合物6(0.7 g，44%)。 (3-乙炔基環戍基）甲醇a)Reagent MW equivalent mmol g, mL Compound 4 130 1.0 31 4.0 g AcCl 78 1.0 34 2.7 g NaH (60°/〇) 24 0.9 28 1.1 g THF 60 mL Stirring cyclopentane-1,3- in an ice bath NaH (60%) (ll g, 28 mol) was added to a solution of diyldiethanol 4 (4.0 g, 31 mmol) in THF (60 mL). The reaction mixture was then stirred under an ice bath for 0.5 hr then ethyl acetate (2.7 g, &lt The mixture was stirred overnight at room temperature. After this time, the mixture was diluted with water (1 mL, EtOAc) NMR (400 MHz, DMSO-A) 0.84-0.87 (m, 1H), 1.28-1.31 (m, 2H), 1.62-1.65 (m, 2H), 1.80-1.83 (m, 1H), 1.99 (s, 1H) ), 2.10-2.18 (m, 1H), 3.26-3.29 (m, 2H), 3.89 (d, ·7=8, 2H), 4.43 (s, 1H). Acetic acid (3-methylindenylcyclopentyl)f ester (6) Reagent MW equivalent mmol g , mL Compound 5 172 1 9.9 1-7 g PCC 215 3.0 29.7 6.4 g CH2C12 50 ml -88· 159849.doc s 201249786 PCC (6.4 g, 29.7 mmol) was added portionwise with a solution of EtOAc (EtOAc (EtOAc) The reaction mixture was then stirred at room temperature overnight. After the mixture was concentrated, the residue was purified EtOAcjjjjjjj (3-ethynylcyclodecyl)methanol a)

試劑 MW 當量 mmol g，mL 化合物6 170 1.0 4.1 0.7 g 貝斯特曼試劑(80%) 192 1.5 6.15 1.18g K2C〇3 138 1.5 6.15 0.85 g MeOH 50 mL 向攪拌之乙酸（3-甲醯基環戊基）曱酯6(0.7 g，4.1 mmol) 於MeOH(50 mL)中之溶液中添加碳酸鉀（0·85 g，6.15 mmol)及貝斯特曼試劑（1 · 18 g，6· 15 mmol)。在室溫下授拌反應混合物隔夜。此後，用水（50 mL)稀釋混合物，用 ***（80 mL><3)萃取且乾燥經合併之有機層，用矽膠層析 (PE:EA=2:1)純化濃縮之殘餘物，得到呈黃色油狀之化合物 7(0.31 g，60%) ° 4-((3-(羥甲基）環戊基）丁 -1，3-二炔基）苯甲酸甲醋 (ACHL02-149)Reagent MW equivalent mmol g, mL Compound 6 170 1.0 4.1 0.7 g Bestmann's reagent (80%) 192 1.5 6.15 1.18g K2C〇3 138 1.5 6.15 0.85 g MeOH 50 mL Agitated acetic acid (3-methylpyridylcyclopentane) Addition of potassium carbonate (0·85 g, 6.15 mmol) and Bestman's reagent (1 · 18 g, 6·15 mmol) to a solution of carbaryl 6 (0.7 g, 4.1 mmol) in MeOH (50 mL) . The reaction mixture was allowed to stand overnight at room temperature. After that, the mixture was diluted with water (50 mL), EtOAc (EtOAc) (EtOAc (EtOAc) Yellow oily compound 7 (0.31 g, 60%) ° 4-((3-(Hydroxymethyl)cyclopentyl)butyl-1,3-diynyl)benzoic acid methyl vinegar (ACHL02-149)

試劑 MW 當量 mmol g，mL 化合物7 124 1.0 2.42 0.3 g 化合物8 240 1.2 2.9 0.7 g PdCl2(PPh3)2 701 0.03 0.1 70 mg Cul 191 0.1 0.242 46 mg DIPEA 129 3.0 7.26 936 g THF 60 mL 159849.doc •89· 201249786 在氮氣下，在室溫下將DIPEA(93 6 mg，7.26 mmol)逐滴添加至（3-乙炔基環戊基）曱醇7(0.3 g，2.42 mmol)、4-(溴乙炔基）苯甲酸甲酯 8(0.7 g，2.9 mmol)、PdCl2(PPh3)2(70 mg，0.1 mmol)及 Cul(46 mg，0.242 mmol)於 THF(60 mL)中之混合物中。在室溫下攪拌反應混合物隔夜。LCMS監測反應。用水（60 mL)稀釋反應混合物且用乙酸乙酯（80 mLx3)萃取。用飽和NH4C1水溶液、水及鹽水洗滌乙酸乙酯層。乾燥之後，減壓濃縮乙酸乙酯溶液。用矽膠管柱 (PE:EA=5:1)純化殘餘物，獲得化合物ACHL02-149(呈順式與反式異構體之混合物）（140 mg，25%)。H NMR (400 MHz, DMSO〇: 7.95 (d，J=8.4 Hz, 2Η)，7.51 (d，J=8 Ηζ， 2H), 3.91 (s, 1H), 3.61 (d, J=4 Ηζ,ΙΗ), 3.53 (d, J=8 Hz, 1H), 2.89-2.87 (m, 1H), 2.43-2.40 (m, 1H), 2.21-2.17 (m, 1H)，2.02-1.99 (m, 1H), 1.82-1.76 (m, 1H)，1.54-1.45 (m，lH )，1.36-1.31 (m，lH)。Reagent MW equivalent mmol g, mL Compound 7 124 1.0 2.42 0.3 g Compound 8 240 1.2 2.9 0.7 g PdCl2(PPh3)2 701 0.03 0.1 70 mg Cul 191 0.1 0.242 46 mg DIPEA 129 3.0 7.26 936 g THF 60 mL 159849.doc • 89· 201249786 DIPEA (93 6 mg, 7.26 mmol) was added dropwise to (3-ethynylcyclopentyl)nonanol 7 (0.3 g, 2.42 mmol), 4-(bromoacetylene) at room temperature under nitrogen. A mixture of methyl benzoate 8 (0.7 g, 2.9 mmol), PdCl 2 (PPh3) 2 (70 mg, 0.1 mmol) and EtOAc (46 mg, 0.242 mmol) in THF (60 mL). The reaction mixture was stirred at room temperature overnight. The reaction was monitored by LCMS. The reaction mixture was diluted with water (60 mL) andEtOAc The ethyl acetate layer was washed with a saturated aqueous NH4Cl solution, water and brine. After drying, the ethyl acetate solution was concentrated under reduced pressure. The residue was purified with a silica gel column (PE: EA = 5:1) to afford Compounds ssssssssssssssssssssssssssssssssssssssss H NMR (400 MHz, DMSO 〇: 7.95 (d, J = 8.4 Hz, 2 Η), 7.51 (d, J = 8 Ηζ, 2H), 3.91 (s, 1H), 3.61 (d, J=4 Ηζ, ΙΗ ), 3.53 (d, J=8 Hz, 1H), 2.89-2.87 (m, 1H), 2.43-2.40 (m, 1H), 2.21-2.17 (m, 1H), 2.02-1.99 (m, 1H), 1.82-1.76 (m, 1H), 1.54-1.45 (m, lH), 1.36-1.31 (m, lH).

將4-((3-(羥甲基）環戊基）丁-i,3 _二炔基）苯甲酸曱酯（115 mg，0.407 mmol，1.0當量）溶解於MeOH(5 mL)中，之後添加氫氧化鋰（29.3 mg，1·222 mmol，3·0 當量）及水（5 mL)。在室溫下攪拌此混合物約72小時。濃縮反應物以移除 MeOH，用6 N HC1(約5 mL)酸化pH值至約3。用EtOAc (3x50 mL)萃取，合併有機層，用飽和Naci洗滌，乾燥 (MgSCU) ’過濾’濃縮。得到〇 14 8粗4_((3_(羥甲基）環戊 159849.doc •90- 201249786 基）丁-1，3·二炔基）苯甲酸。lCMS M+l，預期值=269.1，觀測值=269.1。Ethyl 4-((3-(hydroxymethyl)cyclopentyl)butan-i,3-diynyl)benzoate (115 mg, 0.407 mmol, 1.0 eq.) was dissolved in MeOH (5 mL) Lithium hydroxide (29.3 mg, 1.222 mmol, 3.0 eq.) and water (5 mL) were added. The mixture was stirred at room temperature for about 72 hours. The reaction was concentrated to remove MeOH and acidified to pH 3 with 6 N EtOAc (~ 5 mL). Extracted with EtOAc (3x 50 mL). 〇 14 8 粗 4_((3_(Hydroxymethyl)cyclopentane 159849.doc •90- 201249786 yl)-butyl-1,3·diynyl)benzoic acid was obtained. lCMS M+l, expected value = 269.1, observed = 269.1.

將（S)-2-胺基-3-(第三丁氧羰基胺基）-3-甲基丁酸甲酯 (100 mg，0.406 mmol，ΐ·〇當量）及 4-((3-(羥曱基）環戊基）丁 -1,3-二炔基）苯甲酸（粗物質）（理論1〇9 mg，0.406 mmol ’ 1.0當量）溶解於dmF(1 mL)中。向此添加DIPEA (0·177 mL，1.016 mmol，2.5 當量），接著添加 ΗΑΤϋ(185 mg，0·488 mmol ’ 1.2當量）。在室溫下攪拌此混合物約16 小時。使反應物於1 Μ檸檬酸與乙酸乙酯之間分配。用半飽和氯化鈉、飽和碳酸氫鈉，接著用飽和氯化鈉洗滌有機物，經硫酸鎂乾燥且蒸發至乾燥。得到50〇 mg粗（S)-3-(第三丁氧羰基胺基）-2-(4-((3-(羥甲基）環戊基）丁-1，3-二炔基）苯甲醯胺基）-3-甲基丁酸曱酯。[CMS M+1，預期值= 497.3，觀測值=497.2。Methyl (S)-2-amino-3-(t-butoxycarbonylamino)-3-methylbutanoate (100 mg, 0.406 mmol, ΐ·〇 equivalent) and 4-((3- Hydroxymercapto)cyclopentyl)butan-1,3-diynyl)benzoic acid (crude) (theoretical 1 〇 9 mg, 0.406 mmol '1.0 eq.) was dissolved in dmF (1 mL). To this was added DIPEA (0·177 mL, 1.016 mmol, 2.5 eq.) followed by hydrazine (185 mg, <RTIgt; The mixture was stirred at room temperature for about 16 hours. The reaction was partitioned between 1 EtOAc and EtOAc. The organics were washed with EtOAc (aq.) 50 mg of crude (S)-3-(t-butoxycarbonylamino)-2-(4-((3-(hydroxymethyl)cyclopentyl)butane-1,3-diynyl)benzene Mercapto)-3-methylbutyrate decyl ester. [CMS M+1, expected value = 497.3, observed = 497.2.

將（2S)-3-(第三丁氧羰基胺基）_2_(4_((3·(羥甲基）環戊基）丁 -1，3-二炔基）苯曱醯胺基）-3 -曱基丁酸甲酯（粗物質）溶解於甲醇（1 mL)中。在室溫下添加含々.ο μ HC1之二噁烷 (0·857 mL ’ 3·43 mmol，8.43當量）。在3小時之後反應完成（HPLC)。在0°C下於旋轉蒸發器上濃縮反應物。向此添加IPA(1 mL)，接著添加羥胺溶液（〇 537 mL，814 mmol， 159849.doc •91· 201249786 20當量）。將燒瓶置於4°C下168小時。反應物濃縮（使反應物保持在0°C下）成膠黏物質。向此添加水（3 mL)及 ACN(0.5 mL)。在0°C下使用TFA(3 mL)酸化。再添加水（1 mL)及 ACN( 1 mL)。藉由 RP HPLC( 1"管柱，20 mL/min，含 0.1% TFA之水/ACN，在5% B下平衡）純化。使用注射過濾器（2x8 mL)加載於1"官柱（1〇 mL/min，5% B)上。經1分鐘勻變至20 mL/rnin。經80分鐘5%至19〇/〇 B，在19〇/〇 B下產物溶離。合併所需溶離份’冷;東且置於冷;東乾燥器(2S)-3-(Tertidinoxycarbonylamino)_2_(4_((3(hydroxymethyl))cyclopentyl)butan-1,3-diynyl)phenylhydrazinyl)-3 Methyl decylbutanoate (crude) was dissolved in methanol (1 mL). Dioxane containing 々.ο μ HC1 (0·857 mL '3·43 mmol, 8.43 eq.) was added at room temperature. The reaction was completed after 3 hours (HPLC). The reaction was concentrated on a rotary evaporator at 0 °C. IPA (1 mL) was added thereto followed by a hydroxylamine solution (〇 537 mL, 814 mmol, 159849.doc • 91· 201249786 20 equivalents). The flask was placed at 4 ° C for 168 hours. The reactants were concentrated (the reaction was maintained at 0 ° C) to form a sticky material. Water (3 mL) and ACN (0.5 mL) were added thereto. Acidified using TFA (3 mL) at 0 °C. Additional water (1 mL) and ACN (1 mL) were added. Purified by RP HPLC (1 "column, 20 mL/min, water containing 0.1% TFA/ACN, equilibrated at 5% B). A syringe filter (2 x 8 mL) was used to load on a 1" column (1 〇 mL/min, 5% B). Change to 20 mL/rnin in 1 minute. The product was dissolved at 19 〇 / 〇 B over 5% to 19 〇 / 〇 B over 80 minutes. Combine the required dissolved fractions 'cold; east and place in cold; east dryer

(m, 1H), 3.21-3.38 (m, 4H), 4.52 (br, 3H), 2.83-2.91 叫，4.65-4.67 (d， 1H), 7.61-7.63 (d, 2H), 7.88-7.90 (d, 2H)5 7.97 (s, 2H) 8.53-8.56 (d，1H)，9.21 (br，1H)，u.19 (s，1H)。 ’ L.合成N-((S)-3-胺基羥胺基）·3_曱基 -1-側氧基丁 -2- 基）·Μ((1，4-順）_4侦甲基）環己基）丁 -13二炔基）苯甲醜胺（1-15)及N-((S)-3-胺基(羥胺基）_3_甲基基）-4-(((1,4-反）-4-(羥甲基）環己基）丁·13_ 胺（1-16) 〒基 -1-側氧基丁二炔基）苯甲醯 159849.doc •92· 201249786(m, 1H), 3.21-3.38 (m, 4H), 4.52 (br, 3H), 2.83-2.91, 4.65-4.67 (d, 1H), 7.61-7.63 (d, 2H), 7.88-7.90 (d , 2H)5 7.97 (s, 2H) 8.53-8.56 (d, 1H), 9.21 (br, 1H), u.19 (s, 1H). 'L. Synthesis of N-((S)-3-aminohydroxylamino)·3_mercapto-1-yloxybutan-2-yl)·Μ((1,4-cis)_4 detective methyl) Cyclohexyl)butyr-13diynyl)benzamide (1-15) and N-((S)-3-amino(hydroxylamino)_3_methyl)-4-(((1,4) -trans)-4-(hydroxymethyl)cyclohexyl)butane 13_amine (1-16) decyl-1-yloxybutadiynyl)benzamide 159849.doc •92· 201249786

DIPEA, THF $ 步驟1 :環己烷-1，4-二甲酸二甲酯（2)DIPEA, THF $ Step 1: Cyclohexane-1,4-dicarboxylic acid dimethyl ester (2)

試劑 MW 當量 mmol g，mL 化合物1 172 1.0 290.7 50 g S0C12 119 4.0 1162.8 138 g MeOH 500 mL 於冰鹽浴中冷卻甲醇（500 mL)且逐滴添加亞硫醯氣（138 g，4.0當量）。向HC1於曱醇中之所得溶液中添加環己烷-1，4-二甲酸1(50 g，1_0當量）且反應混合物回流1小時。 TLC監測反應。蒸發溶劑且用水稀釋殘餘物，用乙酸乙酯萃取。用NaHC03溶液、鹽水洗滌乙酸乙酯層且經Na2S04 乾燥。蒸發溶劑獲得目標化合物（56.4 g，97%)。步驟2 ··環己烷-1，4-二基二甲醇（3) 試劑 MW 當量 mmol g ' mL 化合物2 200 1 150.0 30 g LiAlH4 38 3.0 450.0 17.1 g THF 200 ml 在0°C下，向LiAlH4於THF中之冰冷懸浮液中逐滴添加環己烷-1,4-二曱酸二甲酯2之溶液。反應混合物在室溫下持續攪拌30分鐘。TLC監測反應。添加12當量水（32.4 mL)以淬滅反應。過濾反應混合物，且濃縮濾液。用Et20稀釋殘 159849.doc •93· 201249786 餘物，經Na2S04乾燥。移除Et20以收集目標化合物，其未經進一步純化即用於下一步驟（19.9 g，92%)。步驟3 :乙酸（4-(羥甲基）環己基> 甲酯（4>Reagent MW Equivalent mmol g, mL Compound 1 172 1.0 290.7 50 g S0C12 119 4.0 1162.8 138 g MeOH 500 mL The methanol (500 mL) was cooled in an ice-salt bath and sulphur sulphur (138 g, 4.0 eq.) was added dropwise. To the resulting solution of HCl in decyl alcohol was added cyclohexane-1,4-dicarboxylic acid 1 (50 g, 1-0 eq.) and the reaction mixture was refluxed for 1 hour. The reaction was monitored by TLC. The solvent was evaporated and the residue was diluted with H. The ethyl acetate layer was washed with a NaHCO.sub.3 solution, brine and dried over Na2SO. The solvent was evaporated to give the title compound (56.4 g, 97%). Step 2 ··cyclohexane-1,4-diyldimethanol (3) Reagent MW equivalent mmol g ' mL Compound 2 200 1 150.0 30 g LiAlH4 38 3.0 450.0 17.1 g THF 200 ml At 0 ° C, to LiAlH4 A solution of cyclohexane-1,4-dicarboxylate 2 was added dropwise to the ice-cold suspension in THF. The reaction mixture was continuously stirred at room temperature for 30 minutes. The reaction was monitored by TLC. 12 equivalents of water (32.4 mL) were added to quench the reaction. The reaction mixture was filtered, and the filtrate was concentrated. Dilute the residue with Et20 159849.doc •93· 201249786 Residue, dried by Na2S04. The Et20 was removed to afford the title compound which was used in the next step (19.9 g, 92%). Step 3: Acetic acid (4-(hydroxymethyl)cyclohexyl> Methyl ester (4>

試劑 MW 當量 mmol g，mL 化合物3 144 1.0 138.2 19.9 g AcCl 78 1.0 138.2 10.78 g NaH(60%) 24 1.0 138.2 5.53 g THF 150 mL 在0°C下向NaH及環己烷-1,4-二基二曱醇3於THF中之冰冷懸浮液中逐滴添加AcCM。反應物在15°C下持續攪拌3小時。TLC監測反應。添加1當量H20(2.5 mL)以淬滅反應。過濾反應混合物，且用乙酸乙酯稀釋濾液。用鹽水（5 mL) 洗滌乙酸乙酯層，經Na2S04乾燥。減壓移除溶劑。用矽膠管柱（DCM:MeOH=50:l至10:1)純化殘餘物獲得目標化合物 (8.6 g，33%)。步驟4 :乙酸（4-甲醯基環己基 > 甲酯（5) 試劑 MW 當量 mmol g，mL 化合物4 186 1.0 46.24 8.6 g (C0C1)2 128 2.0 92.47 11.8g DMSO 78 4.0 184.95 14.4 g DCM 200 mL TEA 101 8.0 369.92 37.36 g 在-78°C下向乙二醯氣於DCM(60 mL)中之溶液中逐滴添加含DMSO之DCM(60 mL)。30分鐘之後，在-78°C下逐滴添加含乙酸（4-(羥甲基）環己基）曱酯4之DCM(80 mL)。在 -78°C下再攪拌反應混合物2小時，接著在-78°C下逐滴添加 159849.doc -94- 201249786 TEA以淬滅反應。使反應混合物升溫至室溫。用DCM稀釋反應混合物，且用飽和氯化銨水溶液及鹽水洗滌。乾燥 DCM層且濃縮。用矽膠管柱（DCM)純化殘餘物，獲得目標化合物（9.0 g，仍因二氣甲烷而潤濕）。步驟5 : (4-乙炔基環己基）甲醇（6)Reagent MW equivalent mmol g, mL Compound 3 144 1.0 138.2 19.9 g AcCl 78 1.0 138.2 10.78 g NaH (60%) 24 1.0 138.2 5.53 g THF 150 mL NaH and cyclohexane-1,4-di in 0 °C AcCM was added dropwise to the ice-cold suspension of bis-sterol 3 in THF. The reaction was stirred at 15 ° C for 3 hours. The reaction was monitored by TLC. One equivalent of H20 (2.5 mL) was added to quench the reaction. The reaction mixture was filtered, and the filtrate was diluted with ethyl acetate. The ethyl acetate layer was washed with brine (5 mL) and dried over Na2EtOAc. The solvent was removed under reduced pressure. The residue was purified with EtOAc EtOAc (EtOAc:EtOAc: Step 4: Acetic acid (4-carboxycyclohexyl) > Methyl ester (5) Reagent MW equivalent mmol g, mL Compound 4 186 1.0 46.24 8.6 g (C0C1)2 128 2.0 92.47 11.8g DMSO 78 4.0 184.95 14.4 g DCM 200 </ RTI> </ RTI> <RTIgt; DCM (80 mL) containing (4-(hydroxymethyl)cyclohexyl) decyl acetate 4 was added dropwise. The reaction mixture was further stirred at -78 °C for 2 hours, then 159849 was added dropwise at -78 °C. .doc -94 - 201249786 TEA to quench the reaction. The reaction mixture was allowed to warm to room temperature. The reaction mixture was diluted with DCM and washed with saturated aqueous ammonium chloride and brine, dried and evaporated. The residue was purified to give the title compound (9.0 g, which was still weighed by di-methane). Step 5: (4-ethynylcyclohexyl)methanol (6)

試劑 MW 當量 mmol g，mL 化合物5 184 1.0 46.2 8.5 g 貝斯特曼試劑(80%) 192 1.3 60.0 11.56g K2C〇3 138 3.0 138.6 19.04 g MeOH 100 mL 在0°C下向乙酸（4-甲醯基環己基）甲酯5(8.5 g，1.0當量）、K2CO3(19.04 g，3.0 當量）於 CH3OH(100 mL)中之混合物中添加貝斯特曼試劑（11.56 g，1.3當量，80°/〇)。將混合物在室溫下攪拌1 5小時。TLC監測反應。用水稀釋混合物，用乙酸乙酯萃取，用鹽水洗滌，Na2S04乾燥，減壓濃縮。用矽膠管柱（PE:EA=3:1)純化粗物質，獲得目標化合物（2.6 g，40%)。步驟6 : 4-((4-(羥甲基）環己基）丁-1，3-二炔基）苯甲酸甲醋 (8)Reagent MW equivalent mmol g, mL Compound 5 184 1.0 46.2 8.5 g Bestmann's reagent (80%) 192 1.3 60.0 11.56g K2C〇3 138 3.0 138.6 19.04 g MeOH 100 mL To acetic acid (4-methyl hydrazine at 0 ° C Add a Bestmann reagent (11.56 g, 1.3 eq, 80°/〇) to a mixture of K2CO3 (19.04 g, 3.0 eq.) in CH3OH (100 mL). . The mixture was stirred at room temperature for 15 hours. The reaction was monitored by TLC. The mixture was diluted with EtOAc (EtOAc)EtOAc. The crude material was purified using a silica gel column (PE: EA = 3:1) to afford the desired compound (2.6 g, 40%). Step 6: 4-((4-(Hydroxymethyl)cyclohexyl)butan-1,3-diynyl)benzoic acid methyl vinegar (8)

試劑 MW 當量 mmol g * mL 化合物6 138 1.0 10.14 1.4 g 化合物7 240 1.2 12.17 2.9 g PdCl2(PPh3)2 701 0.03 0.304 213 mg Cul 191 0.1 1.014 194 mg DIPEA 129 3.0 30.43 3.92 g THF 150 mL 159849.doc -95- 201249786 在氮氣下，在室溫下將DIPEA(3.92 g，3.0當量）逐滴添加至（4-乙炔基環己基）曱醇6(丨4 g，U當量）、4-(溴乙炔基）苯甲酸甲酯 7(2.9 g，1.2當量）、PdCl2(PPh3)2(213 mg， 0.03 當量）及 Cul(194 mg，〇_1 當量）於 THF(150 mL)中之混合物中。在室溫下攪拌反應混合物隔夜。lcms監測反應。用水（100 mL)稀釋反應混合物且用乙酸乙酯（300 mL) 萃取。用飽和NH4C1水溶液、水及鹽水洗滌乙酸乙酯層。乾燥之後’減壓濃縮乙酸乙酯溶液。用矽膠管柱（pe:ea= 5:1至3:1)純化殘餘物獲得目標化合物（呈順式與反式異構體之混合物，l·2g，40%)。1HNMR(400 MHz，DMSO-d6)\ 7.959-7.937 (d, J=8.8 Hz, 2H), 7.682-7.660 (d, J=8.8 Hz, 2H), 4.421-4.395(t, J=5.2 Hz, 1H), 3.860(s.3H), 3.207-3.179(t, J=5.6 Hz, 2H), 2.085 (s, 1H), 1.980-1.941 (m, 2H), 1.747-1.708 (m, 2H), 1.366-1.328 (m, 3H), 0.935-0.897 (m, 2H)。Reagent MW equivalent mmol g * mL Compound 6 138 1.0 10.14 1.4 g Compound 7 240 1.2 12.17 2.9 g PdCl2(PPh3)2 701 0.03 0.304 213 mg Cul 191 0.1 1.014 194 mg DIPEA 129 3.0 30.43 3.92 g THF 150 mL 159849.doc - 95- 201249786 DIPEA (3.92 g, 3.0 eq.) was added dropwise to (4-ethynylcyclohexyl)nonanol 6 (丨4 g, U equivalent), 4-(bromoethynyl) at room temperature under nitrogen. A mixture of methyl benzoate 7 (2.9 g, 1.2 eq.), PdCl2 (PPh3) 2 (213 mg, 0.03 eq.) and C.sub.1 (194 mg, 〇1 eq.) in THF (150 mL). The reaction mixture was stirred at room temperature overnight. Lcms monitors the response. The reaction mixture was diluted with water (100 mL) andEtOAc. The ethyl acetate layer was washed with a saturated aqueous solution of NH4Cl, water and brine. After drying, the ethyl acetate solution was concentrated under reduced pressure. The residue was purified with a silica gel column (pe: ea = 5:1 to 3:1) to afford the title compound (yield of cis and trans isomers, l.2g, 40%). 1H NMR (400 MHz, DMSO-d6)\ 7.959-7.937 (d, J = 8.8 Hz, 2H), 7.682-7.660 (d, J = 8.8 Hz, 2H), 4.421-4.395 (t, J = 5.2 Hz, 1H ), 3.860(s.3H), 3.207-3.179(t, J=5.6 Hz, 2H), 2.085 (s, 1H), 1.980-1.941 (m, 2H), 1.747-1.708 (m, 2H), 1.366- 1.328 (m, 3H), 0.935-0.897 (m, 2H).

159849.doc •96- 201249786159849.doc •96- 201249786

4-((4-(羥甲基）環己基）丁-1,3-二炔基）苯甲酸(2) 試劑 MW 當量 Mmol g，mL 化合物1 296 1 4.1 1.2 g Li0H.H20 42 3 12.3 517 mg THF 20 mL H20 5 mL 向4-((4-(羥曱基）環己基）丁-1，3-二炔基）苯甲酸曱酯1(1.2 g，4.1 mmol)於THF(20 mL)中之溶液中添加單水合氫氧化經（5 1 7 mg，12.3 mmol)於水（5 mL)中之溶液。該混合物在室溫下持續攪拌隔夜。此後，減壓移除溶劑，用水（40 mL)稀釋殘餘物，用1 M HC1將pH值調節至4.0，接著用乙酸乙酯（3 x60 mL)萃取。乾燥有機層且濃縮得到呈黃色固體狀之4-((4-(羥甲基）環己基）丁-1,3-二炔基）苯曱酸2(1.0 g，93%)，其未經進一步純化即用於下一步驟。LC-MS:283 [M+H]+。 (S)-3-(第三丁氧羰基胺基）-2-(4-((4-(羥甲基）環己基）丁-l,h二炔基）苯甲醯胺基）-3-甲基丁酸甲酯（3)4-((4-(Hydroxymethyl)cyclohexyl)butan-1,3-diynyl)benzoic acid (2) Reagent MW Equivalent Mmol g, mL Compound 1 296 1 4.1 1.2 g Li0H.H20 42 3 12.3 517 Mg THF 20 mL H20 5 mL to decyl 4-((4-(hydroxyindenyl)cyclohexyl)butan-1,3-diynyl)benzoate 1 (1.2 g, 4.1 mmol) in THF (20 mL) A solution of (5 1 7 mg, 12.3 mmol) in water (5 mL) was added to the solution. The mixture was continuously stirred overnight at room temperature. Thereafter, the solvent was removed under reduced pressure, and the residue was diluted with water (40 mL) The organic layer was dried <RTI ID=0.0></RTI> to <RTI ID=0.0> Further purification was used in the next step. LC-MS: 283 [M+H]+. (S)-3-(Tertidinoxycarbonylamino)-2-(4-((4-(hydroxymethyl)cyclohexyl)butan-l,hdiynyl)benzimidamide)-3 -methyl methyl butyrate (3)

試劑 MW 當量 mmol g，mL 化合物2 282 1 3.72 1.05 g Boc·二-Mc_DAP 246 1 3.72 915 mg HATU 380 1.1 4.1 1.55 g DIPEA 129 3 11.2 1.44 g DMF 25 mL 向4-((4-(羥曱基）環己基）丁-1，3-二炔基）苯曱酸2(1.05 g，3.72 mmol)、HATU(1.55 g，4_1 mmol)於DMF(25 mL) 中之溶液中添加（S)-2-胺基-3-(第三丁氧羰基胺基）-3-甲基 159849.doc •97- 201249786 丁酸曱酯（915 mg，3_72 mmoL)及 DIPEA(1.44 g, 11·2 mmoL)。混合物在室溫下持續攪拌5小時。此後，用水（2〇 mL)稀釋反應物，接著用乙酸乙酯（6〇 mLx3)萃取。用水’ 接著用鹽水洗滌經合併之有機層，乾燥且濃縮，用石夕膠層析（PE:EA 2:1)純化殘餘物，得到呈黃色固體狀之（s)_3 (第二丁氧羰基胺基）-2-(4-((4-(羥曱基）環己基）丁 _1>3_二炔基）苯曱醯胺基）-3-曱基丁酸甲酯3(16 g，85〇/。）。l〇MS:511 [M+H]+。455 [M-56]。 (S)-L胺基-2-(4-((4-(羥γ基）環己基、τ二快基）笨甲酿胺基W甲基丁酸甲酯鹽酸鹽（4)Reagent MW Equivalent mmol g, mL Compound 2 282 1 3.72 1.05 g Boc·Di-Mc_DAP 246 1 3.72 915 mg HATU 380 1.1 4.1 1.55 g DIPEA 129 3 11.2 1.44 g DMF 25 mL to 4-((4-(hydroxy) Add (S)-2 to a solution of cyclohexyl)butane-1,3-diynyl)benzoic acid 2 (1.05 g, 3.72 mmol), HATU (1.55 g, 4_1 mmol) in DMF (25 mL) -Amino-3-(t-butoxycarbonylamino)-3-methyl 159849.doc •97- 201249786 decyl butyrate (915 mg, 3_72 mmoL) and DIPEA (1.44 g, 11.2 mmoL). The mixture was kept stirring at room temperature for 5 hours. Thereafter, the reaction was diluted with water (2 mL) and then extracted with ethyl acetate (6 mL). The combined organic layer was washed with EtOAc (EtOAc) EtOAc (EtOAc) Amino)-2-(4-((4-(hydroxy)ylcyclohexyl)butanyl)>3-diynyl)phenylphenylamino)-3-mercaptobutyric acid methyl ester 3 (16 g , 85 〇 /.). l〇MS: 511 [M+H]+. 455 [M-56]. (S)-L-amino-2-(4-((4-(hydroxy)yl)cyclohexyl, τ-di- yl), benzyl alcohol, methyl methacrylate, methyl ester (4)

試劑 MW 當量 mmol g , mL 化合物3 510 1 3.14 1.6 g HCl(g) CH3OH 20 mL 將（S)-3-(第三丁氧羰基胺基）-2-(4-((4-(羥f基）環己基）丁-1，3-二炔基）苯甲醯胺基）_3·甲基丁酸甲酯6 g，3 14 mmol)於曱醇（20 mL)中之溶液連接至HC1裝置。接著在室溫下授拌反應物直至TLC指示起始物質完全消耗。之後，減壓濃縮溶液，得到呈黃色固體狀之（S)_3-胺基-2-(4-((4-(經曱基）環己基）丁-1,3-二炔基）苯甲醯胺基）_3_甲基丁酸曱酯鹽酸鹽 4(1.4 g，99%)。LC-MS:411 [M+H]+。藉由對掌性HPLC分離順式及反式異構體得到（s)-3-胺基-2-(4-((( 1，4-順）-4-(羥甲基）環己基）丁 - i，3-二炔基）苯甲醯胺基）_3_甲基丁酸曱酯（ACHL02-148-偶合，90 mg)作為峰1及（S)-3-胺基-2-(4-(((1,4-反）-4-(羥甲基）環己基）丁-i，3-二炔基）苯甲 159849.doc •98· 201249786 醯胺基）-3·曱基丁酸曱酯（ACHL02-147-偶合，750 mg)作為 2 °Reagent MW equivalent mmol g , mL Compound 3 510 1 3.14 1.6 g HCl (g) CH3OH 20 mL (S)-3-(Tertidinoxycarbonylamino)-2-(4-((4-(hydroxyf) Base) cyclohexyl) butyl-1,3-diynyl)benzhydrylamino)_3·methyl methyl butyrate 6 g, 3 14 mmol) solution in decyl alcohol (20 mL) was attached to the HC1 unit . The reaction was then stirred at room temperature until TLC indicated complete consumption of starting material. After that, the solution was concentrated under reduced pressure to give (S)- 3-amino-2-(4-((4-(pyridyl)cyclohexyl)butane-1,3-diynyl) benzene as a yellow solid. Amidino) _3_methylbutyrate decanoate hydrochloride 4 (1.4 g, 99%). LC-MS: 411 [M+H]+. (s)-3-Amino-2-(4-(((1,4-1,4-cis)-4-(hydroxymethyl)cyclohexyl)) was obtained by separation of the cis and trans isomers by HPLC. Butyl-i,3-diynyl)benzhydrylamino)_3_methylbutyrate (ACHL02-148-coupling, 90 mg) as peak 1 and (S)-3-amino-2-( 4-(((1,4-trans)-4-(hydroxymethyl)cyclohexyl)butan-i,3-diynyl)benzene 159849.doc •98· 201249786 amidino)-3·fluorenyl Ethyl butyrate (ACHL02-147-coupling, 750 mg) as 2 °

將（S)-3-胺基-2-(4-(((1,4-反）-4-(羥甲基）環己基）丁]，3· 一块基）本甲酿胺基）-3-曱基丁酸曱醋（750 mg，1,827 mmol，1.0當量）添加至圓底燒瓶中。向此添加IpA(6 mL)，音波處理懸浮液5分鐘，接著添加THF(2 mL)形成溶液。添加經胺溶液（2.4 mL，36.5 mmol，20當量），且將燒瓶置於4°C下約96小時。反應物濃縮（反應物維持在〇它下）以移除IPA及THF。在(TC下使用TFA(4 mL)酸化。再添加 ACN(2 mL)。藉由RP HPLC(2"管柱，50 mL/min，含〇.ι〇/0 TFA之水/ACN，在10% B下平衡）純化。使用注射過渡器 (3x8 mL)加載於2”管柱（10 mL/min，10% B)上。經1分鐘句(S)-3-Amino-2-(4-(((1,4-(trans))-4-(hydroxymethyl)cyclohexyl)], 3)-based) 3-Mercaptobutyrate vinegar (750 mg, 1,827 mmol, 1.0 eq.) was added to a round bottom flask. To this was added IpA (6 mL), and the suspension was sonicated for 5 minutes, then THF (2 mL) was added to form a solution. An amine solution (2.4 mL, 36.5 mmol, 20 eq.) was added and the flask was placed at 4 ° C for about 96 hours. The reaction was concentrated (the reaction was maintained under it) to remove IPA and THF. Acidified with TFA (4 mL) under TC. ACN (2 mL) was added. RP HPLC (2 "column, 50 mL/min, water containing 〇.ι〇/0 TFA/ACN, at 10 Purified by equilibration at % B. Loaded on a 2" column (10 mL/min, 10% B) using a syringe transition (3 x 8 mL).

變至 50 mL/min。經 60分鐘 10%至 40% B，在 30.5%至 35% B 下產物溶離。合併所需溶離份，冷凍且置於冷凍乾燥器上。得到706 mg N-((S)-3-胺基-1-(羥胺基）-3·曱基側氧基丁-2-基）-4-(((1，4·反）-4-(羥曱基）環己基）丁-1，3_二炔基）苯甲醯胺1-16，丁？入。1^河8 1^+1，預期值=412.2，觀測值 =412.2。4 NMR (DMSO-A): δ 0.84-0.94 (m，2H)，1.25 (s， 3H), 1.30 (s, 3H), 1.32-1.36 (m, 1H), 1.68-1.72 (m, 2H), 1.90-1.95 (m, 2H), 2.39-2.42 (m, 1H), 3.15-3.18 (m, 2H), 4.38-4.40 (t, 1H), 4.65-4.67 (d, 1H), 7.62-7.64 (d, 2H), 159849.doc •99- 201249786 7.89-7.91 (d, 2H), 7.98 (br, 2H), 8.54-8.56 (d, 1H), 9.22 (br，1H)，11.21 (br，1H)。Change to 50 mL/min. The product was dissolved at 30.5% to 35% B over 60 minutes from 10% to 40% B. The desired fractions were combined, frozen and placed on a freeze dryer. 706 mg of N-((S)-3-amino-1-(hydroxyamino)-3·indolyloxybutan-2-yl)-4-(((1,4·trans)-4-) (hydroxyl decyl) cyclohexyl) butyl-1,3_diynyl) benzoguanamine 1-16, butyl? In. 1^河8 1^+1, expected value = 412.2, observed value = 412.2. 4 NMR (DMSO-A): δ 0.84-0.94 (m, 2H), 1.25 (s, 3H), 1.30 (s, 3H) , 1.32-1.36 (m, 1H), 1.68-1.72 (m, 2H), 1.90-1.95 (m, 2H), 2.39-2.42 (m, 1H), 3.15-3.18 (m, 2H), 4.38-4.40 ( t, 1H), 4.65-4.67 (d, 1H), 7.62-7.64 (d, 2H), 159849.doc •99- 201249786 7.89-7.91 (d, 2H), 7.98 (br, 2H), 8.54-8.56 ( d, 1H), 9.22 (br, 1H), 11.21 (br, 1H).

將（S)-3-胺基-2-(4-(((1，4-順）-4-(羥甲基）環己基）丁 ·1，3-二炔基）苯甲醯胺基）-3-曱基丁酸曱酯（90 mg，0.219 mmol ’ 1.0當量）添加至圓底燒瓶中。向此添加IPA(〇.5 mL) ’渦動溶液2分鐘，接著添加羥胺溶液（0,29 mL，4.38 mmol，20當量）且將燒瓶置於4°C下約72小時《反應物濃縮 (反應物維持在0°C下）以移除IPA。在0°C下使用TFA(3 mL) 酸化。添加乙腈（0.5 mL) »藉由RP HPLC(1"管柱，25 mL/min ’含〇·ι% TFA之水/ACN，在1〇%Β下平衡）純化。使用注射過濾器（7 mL)加載於1"管柱（1〇 mL/min，10% B) 上。經1分鐘均變至25 mL/min。經90分鐘10%至40% B，在25.5%至28% B下產物溶離。合併所需溶離份，冷束且置於冷凍乾燥器上。得到53 mg N-((S)-3-胺基-1-(羥胺基）_ 3-甲基-1-側氧基丁-2_基）-4-(((1，4-順）-4-(羥曱基）環己基）丁 —炔基）苯甲醯胺1-15，TFA。LCMS M+1，預期值 =412.2’ 觀測值=412·2。NMR (DMSO-A): δ 1.20-1.35 (m，2Η)，1.26 (s，3Η)，1.30 (s，3Η)，1,47-1.59 (m，4Η)， 1.71-1.75 (m5 2H), 2.97-2.99 (m, 1H), 3.21-3.22 (d, 2H), 4.30 (br, 1H), 4.65-4.68 (d, 1H), 7.64-7.66 (d, 2H), 7.89-7-91 (d, 2H), 7.98 (br, 2H), 8.54-8.56 (d, 1H), 9.22 (br, 159849.doc •100- 201249786 1H)，11.20 (s，1H) ° M.合成N-((S)-1-(1-胺基環丁基）_2_(羥胺基）_2側氧基乙基）-4-(((反）-2-(羥甲基）環丙基）丁_i，3_二炔基）苯甲醯胺三氟乙酸鹽（I-17)&N_((S)-1-(1-胺基環丁基）_2_(羥胺基）_2側氧基乙基）-4-(((順）-2-(羥甲基）環丙基）丁-it二炔基）苯甲醢胺三氟乙酸鹽（1-18)(S)-3-Amino-2-(4-(((1,4-cis)-4-(hydroxymethyl)cyclohexyl)butane)-1,3-diynyl)benzamide Ethyl -3-mercaptobutyrate (90 mg, 0.219 mmol '1.0 eq.) was added to a round bottom flask. Add IPA (〇.5 mL) to this vortex solution for 2 minutes, then add hydroxylamine solution (0,29 mL, 4.38 mmol, 20 eq.) and place the flask at 4 ° C for about 72 hours. The material was maintained at 0 ° C) to remove the IPA. Acidified using TFA (3 mL) at 0 °C. Acetonitrile (0.5 mL) was added. Purified by RP HPLC (1 "column, 25 mL/min <RTI ID=0.0>> A syringe filter (7 mL) was used to load on a 1"column (1 〇 mL/min, 10% B). It changed to 25 mL/min in 1 minute. The product was dissolved at 25.5% to 28% B over 90 minutes from 10% to 40% B. The desired fractions were combined, cold bundled and placed on a freeze dryer. Obtained 53 mg of N-((S)-3-amino-1-(hydroxyamino)-3-methyl-1-butoxybutan-2-yl)-4-(((1,4-cis) 4-(hydroxydecyl)cyclohexyl)butynyl)benzamide 1-15, TFA. LCMS M+1, expected = 412.2. observed = 412. NMR (DMSO-A): δ 1.20-1.35 (m, 2 Η), 1.26 (s, 3 Η), 1.30 (s, 3 Η), 1, 47-1.59 (m, 4 Η), 1.71-1.75 (m5 2H), 2.97-2.99 (m, 1H), 3.21-3.22 (d, 2H), 4.30 (br, 1H), 4.65-4.68 (d, 1H), 7.64-7.66 (d, 2H), 7.89-7-91 (d , 2H), 7.98 (br, 2H), 8.54-8.56 (d, 1H), 9.22 (br, 159849.doc •100- 201249786 1H),11.20 (s,1H) ° M.Synthesis N-((S) 1-(1-Aminocyclobutyl)_2-(hydroxylamino)_2 pendant oxyethyl)-4-(((trans)-2-(hydroxymethyl)cyclopropyl)butane _i,3_ Diynyl)benzamide amine trifluoroacetate (I-17) & N_((S)-1-(1-aminocyclobutyl)_2_(hydroxylamino)_2 oxoethyl)-4 -(((cis)-2-(hydroxymethyl)cyclopropyl)butan-it diynyl)benzimidamide trifluoroacetate (1-18)

/0H ^ /0H/0H ^ /0H

將4-(((反）-2-(經甲基）壞丙基）丁- i，3_二快基）苯甲酸（205 mg，0·85 mmol，1當量）及（S)-2-胺基-2-(1-(第三丁氧羰基胺基）環丁基）乙酸甲西旨（220 mg，0.85 mmol，1當量）溶解於二曱基甲醯胺（3 mL)中。添加二異丙基乙胺 (0.45 mL，2.6 mmol，3 當量），之後添加 ΗΑΤϋ(35ό mg， 0.94 mmol ’ 1.1當量）。混合物在室溫下維持2 5小時，接著於1 Μ檸檬酸與乙酸乙酯之間分配。用1 μ檸檬酸、飽和碳酸氫鈉’接著用飽和氯化鈉洗滌有機物，經硫酸鎂乾燥且蒸發至乾燥’得到（S)-2-(l-(第三丁氧羰基胺基）環丁基）_ 2-(4-(((反）-2-(羥曱基）環丙基）丁-1,3-二炔基）苯甲醯胺基）乙酸甲酯（550 mg粗物質’ 134%，未經進一步純化即使用）。4-(((trans)-2-(methyl)-propyl)buty-i,3-di- yl)benzoic acid (205 mg, 0.85 mmol, 1 eq.) and (S)-2 -Amino-2-(1-(t-butoxycarbonylamino)cyclobutyl)acetic acid methyl carbazide (220 mg, 0.85 mmol, 1 eq.) was dissolved in dichloromethane (3 mL). Diisopropylethylamine (0.45 mL, 2.6 mmol, 3 eq.) was added followed by hydrazine (35 ό mg, 0.94 mmol </ s). The mixture was maintained at room temperature for 25 hours and then partitioned between 1 EtOAc and EtOAc. The organics were washed with 1 μ of citric acid, saturated sodium bicarbonate' followed by saturated sodium chloride, dried over magnesium sulfate and evaporated to dryness to give (S)-2-(l-(t-butoxycarbonylamino)cyclohexane. Base) 2-(4-(((trans)-2-(hydroxy)yl)propyl-1,3-1,3-diynyl)benzamide) methyl acetate (550 mg crude material' 134%, used without further purification).

/〇H/〇H

159849.doc -101· 201249786 將（S)-2-(l-(第三丁氧羰基胺基）環丁基）2 (4 (((反）2_ (羥甲基）環丙基）丁_1，3_二炔基）苯甲醯胺基）乙酸曱酯（55〇 mg粗物質，hl mm〇1)溶解於二氣甲烷（6 mL)中且在室溫下用6 mL三氟乙酸處理10分鐘。移除揮發物且自二氯甲烷再蒸發殘餘物，得到呈三氟乙酸鹽形式之02 — 0 •胺基環丁基）-2-(4-(((反）-2-(羥甲基）環丙基）丁·u_二炔基）苯甲醯胺基）乙酸甲酯，其未經進一步純化即使用。159849.doc -101· 201249786 (S)-2-(l-(Tertidinoxycarbonylamino)cyclobutyl)2 (4(((trans)2_(hydroxymethyl)cyclopropyl)) 1,3_Diynyl)benzamide amino) decyl acetate (55 〇mg crude material, hl mm〇1) was dissolved in di-methane (6 mL) and 6 mL trifluoroacetic acid at room temperature Process for 10 minutes. The volatiles were removed and the residue was evaporated from dichloromethane to give <RTI ID=0.0>> Methyl cyclopropyl) butyl u-diynyl) benzhydryl) acetate, which was used without further purification.

將以上反應中所獲得之粗（s)_2_(1_胺基環丁基）2 (4_ (((反）-2·(羥甲基）環丙基）丁-L3-二炔基）苯甲醯胺基）乙酸甲酉曰二氟乙酸鹽溶解於異丙醇（2 mL)中且在4°c下用50%經胺水/谷液（2 mL)處理20小時。移除揮發物且將殘餘物溶解於用二氟乙酸酸化之水/乙腈中。藉由逆相Hplc(2"管柱， 50 mL/min，含〇·ΐ% TFA之水/ACN，在2%下平衡）使用以下梯度純化該物質： 2%，10分鐘 2-15%，5分鐘 15-45%，300分鐘在5 8分鐘與7 8分鐘之間產物溶離，且凍乾所需溶離份以獲得呈三氟乙酸鹽形式之N-((S)-1 -(1-胺基環丁基）-2_(羥胺基）-2-側氧基乙基）-4-(((反）-2-(經甲基）環丙基）丁_丨，3_二炔 159849.doc -102- 201249786 基）苯甲離胺（白色固體，120 mg，0.24 mmo卜自4-(((反）_ 2-(羥甲基）環丙基）丁 _i，3_二炔基）笨甲酸為28%)。質譜資料：預期值（M+1) : 382.4 ’觀測值：382」。質子 (400 MHz, dmso-^)： H.3 (br s, 1H), 9.24 (s, 1H), 8.62 (d, 1H，J=8.8Hz)，8.17 (br s, 3H)，7.86 (d，2H，J=8.4Hz)，7.63 (d, 2H, J=8.4Hz), 4.92 (d, 1H, J=9.2Hz), 4.69 (t, 1H, J=5.8Hz), 3.40 (m, 1H), 3.26 (m, 1H), 2.12-2.21 (m, 4H), 1.89 (m, 1H), 1.80 (m, 1H), 1.39-1.46 (m, 2H), 0.84-0.93 (m，2H)。再純化不純溶離份後再獲得8〇 mg。The crude (s)_2_(1_aminocyclobutyl) 2 (4_((trans)-2.(hydroxymethyl)cyclopropyl)butan-L3-diynyl)benzene obtained in the above reaction The methionine diacetate diacetate acetate was dissolved in isopropanol (2 mL) and treated with 50% aqueous amine/glutamine (2 mL) for 20 hours at 4 °C. The volatiles were removed and the residue was dissolved in water / acetonitrile acidified with difluoroacetic acid. Purify the material by reverse phase Hplc (2 "column, 50 mL/min, water containing 〇·ΐ% TFA/ACN at 2%) using the following gradient: 2%, 10 minutes 2-15%, 15-45% for 5 minutes, the product is dissolved between 5 8 minutes and 7 8 minutes for 300 minutes, and the desired fraction is lyophilized to obtain N-((S)-1 -(1- in the form of trifluoroacetate salt. Aminocyclobutyl)-2_(hydroxylamino)-2-oxoethyl)-4-(((trans)-2-(methyl)cyclopropyl)butanyl, 3-diyne 159849 .doc -102- 201249786 base) Benzene-free amine (white solid, 120 mg, 0.24 mmo from 4-(((trans))-2-(hydroxymethyl)cyclopropyl) butyl, 3-diyne Base) stupid formic acid is 28%). Mass Spectrometry Data: Expected Value (M+1): 382.4 ‘Observation: 382”. Proton (400 MHz, dmso-^): H.3 (br s, 1H), 9.24 (s, 1H), 8.62 (d, 1H, J=8.8Hz), 8.17 (br s, 3H), 7.86 (d , 2H, J=8.4Hz), 7.63 (d, 2H, J=8.4Hz), 4.92 (d, 1H, J=9.2Hz), 4.69 (t, 1H, J=5.8Hz), 3.40 (m, 1H) ), 3.26 (m, 1H), 2.12-2.21 (m, 4H), 1.89 (m, 1H), 1.80 (m, 1H), 1.39-1.46 (m, 2H), 0.84-0.93 (m, 2H). After purifying the insoluble fraction, 8 〇 mg was obtained.

λ—NH O OH 分別將主要含有N-((S)-1-(1-胺基環丁基）_2_(羥胺基）_2_ 側氧基乙基）-4-(((順）-2-(羥甲基）環丙基）丁-1>3_二炔基）苯甲醯胺（主要異構體中之雜質）之溶離份凍乾且在相同梯度下再純化。彙集所需溶離份且凍乾，得到呈三氟乙酸鹽形式之N-((S)-1-(1-胺基環丁基）-2-(經胺基）-2-側氧基乙基）-4-(((順）-2-(羥曱基）環丙基）丁 -1,3-二炔基）苯甲醯胺（3.2 mg，8.4 μηιοί’ 自 4-(((反）-2-(羥甲基）環丙基）丁-二炔基）苯曱酸為1°/〇)。質譜資料：預期值（M+1) : 382.4 ,觀測值· 382.1。質子NMR (400 MHz, dmso-(i6): 11.30 (bs s， 1H), 9.24 (s, 1H), 8.62 (d, 1H, J=9.2 Hz), 8.16 (br s, 3H), 7.86 (d, 2H, J=7.6 Hz), 7.65 (d, 2H, J=7.6 Hz), 4.92 (d, 1H, J=9.2 Hz), 4.70 (t, 1H, J=5.2 Hz), 3.49 (m, 1H), 3.40 (m, 159849.doc 201249786 1H), 2.12-2.22 (m, 4H), 1.82-1.95 (m, 1H), 1.70-1.80 (m, 2H)，1.36 (m，1H)，1.08 (m，1H)，0.58 (m’ 1H)。 Ν·合成N-((S)-l-(l-(二甲胺基）環丁基）_2·(羥胺基）-2-側氧基乙基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁·][，3_二炔基）苯甲醯胺（1-19)λ-NH O OH will mainly contain N-((S)-1-(1-aminocyclobutyl)_2_(hydroxylamino)_2_ side oxyethyl)-4-(((cis)-2-) The soluble fraction of (hydroxymethyl)cyclopropyl)butan-1>3-dipropargyl)benzamide (impurity in the major isomer) was lyophilized and repurified under the same gradient. The desired fractions are pooled and lyophilized to give N-((S)-1-(1-aminocyclobutyl)-2-(amino)-2-oxoethoxy b as a trifluoroacetate salt. Base)-4-(((cis)-2-(hydroxyindenyl)cyclopropyl)butan-1,3-diynyl)benzamide (3.2 mg, 8.4 μηιοί' from 4-(((反-2-(hydroxymethyl)cyclopropyl)butan-diynyl)benzoic acid is 1°/〇). Mass spectral data: expected value (M+1): 382.4, observed · 382.1. Proton NMR (400 MHz, dmso-(i6): 11.30 (bs s, 1H), 9.24 (s, 1H), 8.62 (d, 1H, J=9.2 Hz), 8.16 (br s, 3H), 7.86 (d , 2H, J=7.6 Hz), 7.65 (d, 2H, J=7.6 Hz), 4.92 (d, 1H, J=9.2 Hz), 4.70 (t, 1H, J=5.2 Hz), 3.49 (m, 1H ), 3.40 (m, 159849.doc 201249786 1H), 2.12-2.22 (m, 4H), 1.82-1.95 (m, 1H), 1.70-1.80 (m, 2H), 1.36 (m, 1H), 1.08 (m , 1H), 0.58 (m' 1H). Ν·Synthesis of N-((S)-l-(l-(dimethylamino)cyclobutyl)_2·(hydroxylamino)-2-oxoethyl )-4-(((1,2-trans)-2-(hydroxymethyl)cyclopropyl)butan]][,3-diynyl)benzamide (1-19)

將N-((S)-1-(1-胺基環丁基）_2_(羥胺基）_2_側氧基乙基）_ 4-(((反）-2-(羥甲基）環丙基）丁-丨，3_二炔基）苯甲醯胺三氟乙酸鹽1-17(80 mg ’ 0.16 mmol，1當量）溶解於愚#-二甲基甲酿胺（800 μΙ〇中且在室溫下用三聚甲醛（48.5 mg，1.1 mm〇】’ 10當量）及愚#-二異丙基乙胺（56 ，〇 32 mmo卜2 當量）處理21小時《添加氰基硼氫化鈉（3〇 mg，0.48 mmol，3當量），之後添加曱醇（8〇〇 μί)及乙酸（28 ， 0.48 mmol，3當量）。混合物在室溫下保持3天，接著再添加氰基硼氫化鈉及乙酸（28 pL)。再處於室溫下一天之後，再添加28 pL乙酸。再處於室溫下一天之後，藉由逆相 HPLC(1"管柱，20 mL/min，含 0.1% TFA 之水/乙腈，在2〇/〇下平衡）使用以下梯度純化該物質： 2%，5分鐘 2-18%，5分鐘 18-98% ’ 80分鐘彙集所需溶離份且凍乾得到呈三氟乙酸鹽形式之N-((s)_ 159849.docN-((S)-1-(1-Aminocyclobutyl)_2_(hydroxylamino)_2_sideoxyethyl)-4-(((trans)-2-(hydroxymethyl)cyclopropane Butyl-indole, 3_diynyl) benzalkonium trifluoroacetate 1-17 (80 mg '0.16 mmol, 1 eq.) was dissolved in ## dimethyl ketoamine (800 μΙ〇 and Treated with trioxane (48.5 mg, 1.1 mm 〇) '10 eq.) and 愚 #-diisopropylethylamine (56, 〇32 mmo b 2 eq.) for 21 hours at room temperature. Add sodium cyanoborohydride (3 〇 mg, 0.48 mmol, 3 eq.), followed by the addition of decyl alcohol (8 〇〇μί) and acetic acid (28, 0.48 mmol, 3 eq.). The mixture was kept at room temperature for 3 days, followed by the addition of cyanoborohydride. Sodium and acetic acid (28 pL). After one day at room temperature, add 28 pL of acetic acid. After one day at room temperature, by reverse phase HPLC (1 " column, 20 mL/min, containing 0.1% TFA The water/acetonitrile was equilibrated at 2 〇/〇). The material was purified using the following gradient: 2%, 2-18% for 5 minutes, 18-98% for 5 minutes. The desired fractions were pooled in 80 minutes and lyophilized to give three N-((s)_ 159849.doc in the form of fluoroacetate

S •104- 201249786 1-(1-(二甲胺基）環丁基）-2-(經胺基）-2-側氧基乙基）-4-(((1，2-反）-2-(羥甲基）環丙基）丁-13_二炔基）苯曱醯胺（63 mg，0·12 mmol，74%)。質譜資料：預期值（M+1): 410_5 ’ 觀測值：410.2。質子NMR (400 MHz, dmso-d6): 11.29 (s, 1H), 9.72 (br s, 1H), 9.30 (s, 1H), 8.95 (d, 1H, J=9.2 Hz), 7.88 (d, 2H, J=8.4 Hz), 7.61 (d} 2H, J=8.4 Hz), 5.05 (d, 1H, J=9.2 Hz), 4.7 (br s, 1H), 3.40 (dd, 1H, J=5.2, 11.6 Hz), 3.22 (dd, 1H, J=6.4, 11.2 Hz), 2.76 (d, 3H, J=4.4 Hz), 2.68 (d, 3H, J=4.4 Hz), 2.33-2.42 (m, 3H), 1.62-1.78 (m，2H)，1.37-1.46 (m，2H), 0.84-0.92 (m，2H)。〇·合成N-((S)-2-(羥胺基）-i-((S)-嗎啉-3-基）-2-側氧基乙基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺（1-20)S •104- 201249786 1-(1-(Dimethylamino)cyclobutyl)-2-(amino)-2-oxoethyl)-4-((1,2-trans)- 2-(Hydroxymethyl)cyclopropyl)butyr-13-diynyl)benzamide (63 mg, 0. 12 mmol, 74%). Mass spectral data: expected value (M+1): 410_5 ′ observed: 410.2. Proton NMR (400 MHz, dmso-d6): 11.29 (s, 1H), 9.72 (br s, 1H), 9.30 (s, 1H), 8.95 (d, 1H, J=9.2 Hz), 7.88 (d, 2H) , J=8.4 Hz), 7.61 (d} 2H, J=8.4 Hz), 5.05 (d, 1H, J=9.2 Hz), 4.7 (br s, 1H), 3.40 (dd, 1H, J=5.2, 11.6 Hz), 3.22 (dd, 1H, J=6.4, 11.2 Hz), 2.76 (d, 3H, J=4.4 Hz), 2.68 (d, 3H, J=4.4 Hz), 2.33-2.42 (m, 3H), 1.62-1.78 (m, 2H), 1.37-1.46 (m, 2H), 0.84-0.92 (m, 2H). 〇·Synthesis of N-((S)-2-(hydroxylamino)-i-((S)-morpholin-3-yl)-2-yloxyethyl)-4-(((1,2- Trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (1-20)

1.氮甲酸異丁酯， TEA, -15°C 至〇°C , 16小時 2.重氮甲烷，0eC,16小時 77% '1. Isobutyl nitrate, TEA, -15 ° C to 〇 ° C, 16 hours 2. Diazomethane, 0eC, 16 hours 77% '

在-16°c下向預先冷卻之（R)-4-(第三丁氧羰基）嗎啉-3-甲酸1(5 g，0.02162 mol)於無水***（160 mL)中之懸浮液中逐滴添加三乙胺（3.91 mL，0.0281 mol)，且在5分鐘之後’在-15°C下用氣甲酸異丁酯（3.37 mL，0.02600 mol)緩慢處理所得混合物持續30分鐘，接著在-15°C至6。(：下授拌反應混合物16小時（隔夜），LCMS監測反應過程，過濾，用***（30 mLx2)洗滌固體，將濾液轉移至下一步驟。在〇至7C下用重氮曱燒（0 24 mol)於乙縫（480 mL)中之溶液處 I59849.doc -105· 201249786 理上述濾液16小時（隔夜）’接著用冰醋酸（5 mL)、水（5〇 mL)>卒滅反應’且用碳酸氫納（5〇 mLU)、鹽水（50 mL)洗滌有機層，用***（200 mLx2)再萃取水層，經無水硫酸鈉乾燥經合併之有機層，過滤，濃縮得到粗殘餘物，藉由石夕膠層析（含20%乙酸乙酯之石油作為溶離劑）純化，得到 4.56 g呈黃色固體狀之（R)-3-(2-重氮乙醯基）嗎啉_4_曱酸第二丁酯 2(純度 87% ’ LCMS ’ 254 nm)，產率：77%，2個步 ? 1 AgOBz, TEA ?"Ί \ ^NBOC MeOH, 25°C 78% k^NBoc 2 Xe 3 向預先冷卻之（R)-3-(2-重氮乙醯基）嗎啉_4_甲酸第三丁酯 2(4.55 g，17.824 mmol)及苯曱酸銀（25 mg、〇 u mm〇1) 於甲醇（30 mL)中之懸浮液中逐滴添加三乙胺（25〇， 1.78 mmol)，且在25°C下在黑暗中攪拌懸浮液16小時（隔夜，油浴），接著過濾，蒸發濾液至乾燥且藉由矽膠層析 (含10%乙酸乙酯之石油作為溶離劑）純化殘餘物，得到 3.34 g呈無色固體狀之（s)_3_(2•甲氧基_2_側氧基乙基）嗎琳-4-曱酸第三丁酯3，產率：78〇/〇。To a suspension of pre-cooled (R)-4-(t-butoxycarbonyl)morpholine-3-carboxylic acid 1 (5 g, 0.02162 mol) in anhydrous diethyl ether (160 mL) at -16 °C Triethylamine (3.91 mL, 0.0281 mol) was added dropwise, and after 5 minutes, the resulting mixture was slowly treated with isobutyl methacrylate (3.37 mL, 0.02600 mol) at -15 ° C for 30 minutes, then at -15 °C to 6. (: The reaction mixture was stirred for 16 hours (overnight), the reaction was monitored by LCMS, filtered, and the solid was washed with diethyl ether (30 mL×2), and the filtrate was transferred to the next step. Mol) solution in splicing (480 mL) I59849.doc -105· 201249786 The above filtrate was treated for 16 hours (overnight)' followed by glacial acetic acid (5 mL), water (5 〇 mL) > The organic layer was washed with EtOAc (EtOAc) (EtOAc (EtOAc) Purification by gas chromatography (20% ethyl acetate as a solvent) gave 4.56 g of (R)-3-(2-diazonylidene) morpholine as a yellow solid. Dibutyl citrate 2 (purity 87% 'LCMS ' 254 nm), yield: 77%, 2 steps? 1 AgOBz, TEA ?"Ί \ ^NBOC MeOH, 25°C 78% k^NBoc 2 Xe 3 pre-cooled (R)-3-(2-diazoethyl)morpholine_4_carboxylic acid tert-butyl ester 2 (4.55 g, 17.824 mmol) and silver benzoate (25 mg, 〇u) Mm〇1) in methanol (30 Triethylamine (25 〇, 1.78 mmol) was added dropwise to the suspension in mL), and the suspension was stirred in the dark at 25 ° C for 16 hours (overnight, oil bath), then filtered and evaporated to dryness. The residue was purified by silica gel chromatography (10% ethyl acetate in EtOAc) to afford 3.34 g of (s) _3_ Lin-4-pyruic acid tert-butyl ester 3, yield: 78 〇 / 〇.

NaHMDS, THF 2,4,6-三異丙基苯磺醢基疊氮 38%NaHMDS, THF 2,4,6-triisopropylbenzenesulfonyl azide 38%

Onboc OMe 在100 c下向預先冷卻之（s)_3_(2_甲氧基_2_側氧基乙基）馬啉4曱酸第三丁酯3(2 737 g，〇〇1〇6爪〇丨）於THF(1〇〇 159849.docOnboc OMe pre-cooled (s)_3_(2_methoxy-2-sideoxyethyl) porphyrin tetradecanoate 3 butyl ester at 100 c (2 737 g, 〇〇1〇6 claws 〇丨) in THF (1〇〇159849.doc

S •106· 201249786 mL)中之溶液中緩慢添加NaHMDS(8.15 mL，0.0159 mol)，且在-100°C下攪拌所得懸浮液1小時，接著緩慢添加 2,4,6-三異丙基苯續醯基疊氮（trisilylazide ; 5.99 g， 0.0170 mol)於THF( 10 mL)中之溶液（維持内部溫度低於-100°C)，且在 -l〇〇°C下再持續攪拌1小時。快速添加冰醋酸（3.18 g，0.0530 mol)以淬滅反應，且使反應混合物升溫至室溫，攪拌3小時，最終用DCM(200 mLx2)稀釋且用飽和 NaHC03 (100 mL)、鹽水（100 mL)及水（100 mL)洗滌。經無水硫酸鈉乾燥經合併之有機層，過濾，濃縮得到粗殘餘物，藉由逆相層析（梯度：含10%乙腈之水歷時5分鐘，在3分鐘内增至含30%乙腈之水，含30%乙腈之水歷時3分鐘，含30%至45%乙腈之水歷時3分鐘，含45%乙腈之水歷時5分鐘；流速：30 mL/min)純化，得到1.22 g呈白色固體狀之（S)-3-(l-疊氮基-2-甲氧基-2-側氧基乙基）嗎啉-4-甲酸第三丁酯4(兩種非對映異構體之混合物），產率：38% όNaHMDS (8.15 mL, 0.0159 mol) was slowly added to the solution in S • 106·201249786 mL), and the resulting suspension was stirred at -100 ° C for 1 hour, followed by slow addition of 2,4,6-triisopropylbenzene A solution of trisilylazide (5.99 g, 0.0170 mol) in THF (10 mL) was maintained (the internal temperature was maintained below -100 °C) and stirring was continued at -1 °C for an additional 1 hour. The glacial acetic acid (3.18 g, 0.0530 mol) was quickly added to quench the reaction, and the reaction mixture was allowed to warm to room temperature, stirred for 3 hours, and finally diluted with DCM (200 mL×2) and saturated NaHC03 (100 mL), brine (100 mL) ) and wash with water (100 mL). The combined organic layers were dried <RTI ID=0.0>(~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Water containing 30% acetonitrile for 3 minutes, water containing 30% to 45% acetonitrile for 3 minutes, water containing 45% acetonitrile for 5 minutes, and flow rate: 30 mL/min to obtain 1.22 g of a white solid. (S)-3-(l-azido-2-methoxy-2-oxoethyl)morpholine-4-carboxylic acid tert-butyl ester 4 (mixture of two diastereomers) ), yield: 38% ό

SMB8-AJ70mg SM68*B, 230mg SM68-C,30mg 將（S)-3-(l-疊氮基-2-甲氧基-2-側氧基乙基）嗎啉-4-甲酸第三丁酯 4(1.2 g，4 mmol)、Pd(OH)2/C(200 mg，15 重量 %)、2 M HC1(2 mL，4 mmol)於 EtOAc/MeOH(20 mL/3 mL) 中之混合物脫氣且在30°C下在1個大氣壓下用氫氣處理16 小時。過濾混合物，且用EtOAc(20 mL)洗滌固體。用水 159849.doc -107- 201249786 (20 mL)、飽和NaHCO3(20 mL，以調節至pH>8)洗膝經合併之有機物，接著經無水硫酸鈉乾燥，過濾，濃縮得到粗殘餘物，藉由逆相層析（梯度：含10%乙腈之水歷時5分鐘，3分鐘内增至含30%乙腈之水，含30%乙腈之水歷時3 分鐘，含30%至45%乙腈之水歷時3分鐘，含45%乙腈之水歷時5分鐘；流速：30 mL/min)純化。接著對純化之殘餘物進行對掌性分離，得到（S)-3-((S)-l-胺基-2-甲氧基-2-側氧基乙基）嗎琳-4-甲酸第二丁醋（SM68-A，170 mg，黃色油狀物）；（S)-3-((R)-l-胺基-2-甲氧基-2-側氧基乙基）嗎琳_ 4-曱酸第三丁酯（SM68-B，230 mg，黃色固體）；及3〇 mg 呈黃色油狀之SM68-C，總產率：41%。SMB8-AJ70mg SM68*B, 230mg SM68-C, 30mg (S)-3-(l-azido-2-methoxy-2-oxoethoxyethyl)morpholine-4-carboxylic acid tert-butyl Mixture of ester 4 (1.2 g, 4 mmol), Pd(OH) 2/C (200 mg, 15% by weight), 2 M HCl (2 mL, 4 mmol) in EtOAc / MeOH (20 mL / 3 mL) Degassed and treated with hydrogen at 1 ° C for 16 hours at 30 °C. The mixture was filtered and the solid was washed with EtOAc EtOAc. Wash the combined organics with water 159849.doc -107 - 201249786 (20 mL), sat. NaHCO3 (20 mL, to pH > 8), then dry over anhydrous sodium sulfate, filtered and concentrated to give a crude residue. Reverse phase chromatography (gradient: water containing 10% acetonitrile for 5 minutes, increased to 30% acetonitrile in 3 minutes, water containing 30% acetonitrile for 3 minutes, water containing 30% to 45% acetonitrile for 3 minutes) In 5 minutes, the water containing 45% acetonitrile was purified for 5 minutes; flow rate: 30 mL/min. The purified residue is then subjected to palm separation to give (S)-3-((S)-l-amino-2-methoxy-2-oxoethyl)m-lin-4-carboxylic acid Dibutyl vinegar (SM68-A, 170 mg, yellow oil); (S)-3-((R)-l-amino-2-methoxy-2-oxoethyl) 4-butyl citrate (SM68-B, 230 mg, yellow solid); and 3 mg of SM68-C as a yellow oil.

將4-(((反）-2-(羥甲基）環丙基）丁 -1，3-二炔基）苯甲酸（140 mg，0.58 mmol)及（S)-3-((S)-l-胺基-2-甲氧基-2-側氧基乙基）嗎啉_4_曱酸第三丁酯（160 mg，0.58 mmol)溶解於 DMF(2 mL)中。添加 TV,#-二異丙基乙胺（255 pL，1.46 mmol) ’ 之後添加 HATU(266 mg，0.70 mmol)。反應在室溫下維持30分鐘，接著於1 檬酸與乙酸乙酯之間分配。用1 Μ檸樣酸、飽和碳酸氫納，接著用飽和氣化鈉洗滌有機物，經硫酸鎂乾燥且蒸發至乾燥。獲得38〇 mg粗 (S)-3-((S)_l-(4-(((l，2-反）-2-(羥甲基）環丙基）丁 _1，3_ 二炔基）苯甲醯胺基）-2·甲氧基-2·側氧基乙基）嗎啉-4-甲酸第三4-(((trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzoic acid (140 mg, 0.58 mmol) and (S)-3-((S) -l-Amino-2-methoxy-2-oxoethylethyl)morpholine_4_decanoic acid tert-butyl ester (160 mg, 0.58 mmol) was dissolved in DMF (2 mL). TV, #-diisopropylethylamine (255 pL, 1.46 mmol) was added followed by HATU (266 mg, 0.70 mmol). The reaction was maintained at room temperature for 30 minutes and then partitioned between 1 citric acid and ethyl acetate. The organics were washed with 1 mL of EtOAc, EtOAc (EtOAc)EtOAc. Obtained 38 mg of crude (S)-3-((S)_l-(4-((l,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-1-, 3-diynyl) Benzylamino)-2·methoxy-2·sideoxyethyl)morpholine-4-carboxylic acid third

159849.doc ·1〇8. S 201249786 丁酯。159849.doc ·1〇8. S 201249786 Butyl ester.

將（S)-3-((S)-l-(4-(((l，2-反）-2-(羥甲基）環丙基）丁-1,3-二炔基）苯甲醯胺基）-2-甲氧基-2-側氧基乙基）嗎啉-4-甲酸第三丁酯（粗物質，380 mg)溶解於二氣甲烷（4 mL)中，添加三氟乙酸（4 mL)。在室溫下5分鐘之後，用10 mL二氣甲烷稀釋混合物且蒸發。由此獲得之（S)-2-(4-(((l,2-反）-2-(羥曱基）環丙基）丁-1，3·二炔基）苯曱醯胺基）-2-((S)-嗎啉-3-基）乙酸甲酯2,2,2-三氟乙酸鹽以粗物質形式用於下一步驟0(S)-3-((S)-l-(4-((l,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzene Tert-butyl)-2-methoxy-2-oxoethylethylmorpholine-4-carboxylic acid tert-butyl ester (crude, 380 mg) was dissolved in di-methane (4 mL) Acetic acid (4 mL). After 5 minutes at room temperature, the mixture was diluted with 10 mL of di-methane and evaporated. (S)-2-(4-((l,2-trans)-2-(hydroxyindenyl)cyclopropyl)butane-1,3·diynyl)phenylhydrazinyl) Methyl 2-((S)-morpholin-3-yl)acetate 2,2,2-trifluoroacetate was used in crude material in the next step.

NH OH 將（S)-2-(4-(((l，2-反）-2-(羥甲基）環丙基）丁 d，3•二炔基）苯曱醯胺基）-2-((S)_嗎啉-3-基）乙酸曱酯2,2,2-三氟乙酸鹽 (來自上述反應之粗物質）溶解於異丙醇（1 mL)中，且添加羥胺水溶液（50%，1 mLp溶液在4°C下維持18小時，接著蒸發。添加水且用TFA酸化混合物。藉由Rp HpLc(2"管柱，50 mL/min ’含〇.1% TFA之水/ACN，在2%下平衡）純化粗物質： ' 2%，10分鐘 2-11%，5分鐘 159849.doc 201249786 11-41%，300分鐘在88分鐘至99分鐘所需化合物溶離。來乾所需溶離份直至得到呈三氟乙酸鹽形式之N-((S)-2-(羥胺基嗎琳-3-基）-2-側氧基乙基）-4-(((1,2-反）-2-(經甲基）環丙基）丁 _ 1，3-二炔基）苯甲醯胺1-20(白色固體，us mg，〇.30 mmol，自4-(((反）-2-(羥甲基）環丙基）丁-丨，3_二炔基）苯曱酸為52%)。質譜資料：預期值（M+1) : 398.2，觀測值： 398.1。質子NMR (400 MHz, dmso-i/<s): 11.17 (s，1H)，9.14 (s, 1H), 9.08 (br s, 1H), 8.93 (br s, 1H), 8.83 (d, 1H, J=8.4 Hz), 7.89 (d, 2H, J=8.0 Hz), 7.62 (d, 2H, J=8.0 Hz), 4.69 (br s, 1H>, 4.63 (t, 1H, J=8.8 HZ), 3.83-3.86 (m, 2H), 3.60-3.66 (m, 2H), 3.50 (t, 1H, J=ll.〇 Hz), 3.39 (m, 1H), 3.20-3.23 (m, 2H), 3.05 (m, 1H), 1.39-1.45 (m, 2H), 0.87-0.92 (m, 2H)。 P.合成N-((S)-2-(羥胺基）-l-((S)-4-甲基嗎啉-3-基）-2-側氧基乙基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁 -1,3-二炔基）苯甲酿胺（1-21)NH OH will be (S)-2-(4-((l,2-trans)-2-(hydroxymethyl)cyclopropyl)butane d,3•diynyl)phenylhydrazinyl)-2 -((S)-morpholin-3-yl)acetate oxime 2,2,2-trifluoroacetate (crude from the above reaction) was dissolved in isopropanol (1 mL), and aqueous hydroxylamine solution was added ( 50%, 1 mLp solution was maintained at 4 ° C for 18 hours, followed by evaporation. Water was added and the mixture was acidified with TFA. By Rp HpLc (2 "column, 50 mL/min 'water containing 1% TFA/ ACN, equilibrated at 2%) to purify the crude material: ' 2%, 10 minutes 2-11%, 5 minutes 159849.doc 201249786 11-41%, 300 minutes in the 88 minutes to 99 minutes required compound dissolution. The fraction is required to be obtained until N-((S)-2-(hydroxyaminophenylin-3-yl)-2-oxoethyl)-4-((1,2-) is obtained as the trifluoroacetate salt. Trans)-2-(methyl)cyclopropyl)butyr-1,3-diynyl)benzamide 1-20 (white solid, us mg, 〇.30 mmol, from 4-((( )-2-(hydroxymethyl)cyclopropyl)butan-indole, 3-diynyl)benzoic acid 52%). Mass spectral data: expected (M+1): 398.2, observed: 398.1. NMR (400 MHz, dmso-i/<s): 11.17 (s, 1H), 9.14 (s, 1H), 9.08 (br s, 1H), 8.93 (br s, 1H), 8.83 (d, 1H, J=8.4 Hz), 7.89 (d, 2H, J= 8.0 Hz), 7.62 (d, 2H, J=8.0 Hz), 4.69 (br s, 1H>, 4.63 (t, 1H, J=8.8 HZ), 3.83-3.86 (m, 2H), 3.60-3.66 (m , 2H), 3.50 (t, 1H, J=ll.〇Hz), 3.39 (m, 1H), 3.20-3.23 (m, 2H), 3.05 (m, 1H), 1.39-1.45 (m, 2H), 0.87-0.92 (m, 2H). P. Synthesis of N-((S)-2-(hydroxylamino)-l-((S)-4-methylmorpholin-3-yl)-2-yloxy Ethyl)-4-(((1,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (1-21)

將N-((S)-2-(羥胺基）-l-((S)-嗎啉-3-基）-2-側氧基乙基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁-1,3-二炔基）苯甲醯胺三氟乙酸鹽1-20(10 mg，20 μηιοί)溶解於DMF(100 pL)中。添加三聚甲醛（5.9 mg，200 μηιοί)，之後添加愚二異丙基 159849.doc -110- 201249786 乙胺（6.8 μί，39 μιηοΐ)。在室溫下攪拌混合物19小時。添加氰基硼氫化鈉（3.7 mg，59 μιηοΐ)、曱醇（1〇〇叫）及乙酸 (4.5 μί ’ 78 μιηοΐ)且在室溫下再攪拌混合物3天。藉由Rp HPLC(2"管柱，50 mL/min ’含〇 1% TFA之水/ACN，在2% 下平衡）純化混合物： 2%，10分鐘 2-15%，5分鐘 15-95%，80分鐘在23分鐘與24分鐘之間產物溶離。凍乾所需溶離份得到呈二氟乙酸鹽形式之N-((S)-2-(羥胺基）_i_((s)_4_甲基嗎啉_ 3·基）-2·側氧基乙基）_4_(((ι，2-反）-2-(羥曱基）環丙基）丁 _ 1，3-一快基）苯曱酿胺（7〇 mg’ η μιη〇ΐ，68%)。質譜資料：預期值（Μ+1) : 411.2，觀測值：412.2。 Q.合成N-((S)-3-胺基-1-經胺基）·3_甲基_1_側氧基丁 _2_ 基）-4-((Ε)-4-(1，2-反）-2-(經甲基）環丙基）丁小稀_3_炔基）苯甲醯胺（1-22)N-((S)-2-(hydroxylamino)-l-((S)-morpholin-3-yl)-2-yloxyethyl)-4-(((1,2-re)) 2-(Hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzimidamide trifluoroacetate 1-20 (10 mg, 20 μηιοί) was dissolved in DMF (100 pL). Add paraformaldehyde (5.9 mg, 200 μηιοί) followed by cumene isopropyl 159849.doc -110- 201249786 ethylamine (6.8 μί, 39 μιηοΐ). The mixture was stirred at room temperature for 19 hours. Sodium cyanoborohydride (3.7 mg, 59 μιηοΐ), decyl alcohol (1 〇〇) and acetic acid (4.5 μί '78 μιηοΐ) were added and the mixture was stirred at room temperature for further 3 days. The mixture was purified by Rp HPLC (2 "column, 50 mL/min 'water containing 1% TFA/ACN, equilibrated at 2%): 2%, 10 minutes 2-15%, 5 minutes 15-95% The product was dissolved between 23 minutes and 24 minutes in 80 minutes. The desired fractions were lyophilized to give N-((S)-2-(hydroxylamino)_i_((s)_4_methylmorpholine-3-yl)-2. ethoxylate B in the form of difluoroacetate. Base)_4_(((ι,2-trans)-2-(hydroxyindenyl)cyclopropyl)butyl 1,3-1,3-fastyl)benzoquinone (7〇mg' η μιη〇ΐ, 68% ). Mass spectral data: expected (Μ +1): 411.2, observed: 412.2. Q. Synthesis of N-((S)-3-aminol-1-amino)·3_methyl_1_sideoxybut-2-yl)-4-((Ε)-4-(1, 2-trans)-2-(methyl)cyclopropyl)butane _3_alkynyl)benzamide (1-22)

NH OH 將（S)-3-胺基-2-(4-(E)-4-(lS,2S)-2-(經甲基）環丙基）丁小稀-3-炔基）苯曱醯胺基）_3_甲基丁酸曱酯（4〇〇 mg，1〇8 mmol，1.0當量）添加至圓底燒瓶中。向此添加異丙醇（2〇 mL)，渦動溶液2分鐘，於冰浴中冷卻燒瓶，接著添加羥胺溶液（1.4 mL，21.60 mmol，20當量）且將燒瓶置於4。〇下約 159849.doc • 111 - 201249786 48小時。濃縮反應物（在〇〇c下）以移除異丙醇，且接著在 0 C使用TFA(3 mL)酸化。再添加水（15 mL)及ACN(3 mL)。藉由逆相HPLC(2"管柱’ 5〇 mL/min，含〇 1% TFA之水/ ACN，在2% B下平衡）純化產物.使用注射過濾器（24 mL) 以10 mL/min，2% B加載管柱，且接著經1分鐘勻變至5〇 mL/min。經73分鐘運作2% B至95。/。B之梯度。合併所需溶離份’冷凍且置於冷凍乾燥器上.反應得到丨77 mg N-((S)-3-胺基-1-(羥胺基）_3-甲基a·側氧基丁 _2_基）_4_((ε)_4· ((1，2-反）2-(羥甲基）環丙基）丁烯_3_炔基）苯曱醯胺^ 22’丁卩八。1^]^]^+1，預期值=372,2，觀測值=3 72.2。4 NMR (DMSO-^6): δ 0.77-0.81 (m, 2H), 1.25 (s, 3H), 1.30 (s, 3H), 1.34-1.37 (m, 1H), 3.20-3.24 (dd, 1H), 3.37-3.41 (dd, 1H), 4.65-4.67 (d, 1H), 6.43-6.47 (d, 1H), 6.87-6.91 (d, 1H), 7.55-7.57 (d, 2H), 7.84-7.86 (d, 2H), 7.99 (br, 2H), 8.38-8.41 (d，lH)，9.22 (br, 1H)，11.20 (s，1H)。 R.抗微生物活性細菌篩選及培養在35°C下於環境空氣中經米勒希爾頓瓊脂（1^1^141·-Hinton agar)(Beckton Dickinson，Franklin Lakes, NJ)由 -70°C冷凍儲備液培養細菌分離株隔夜繼代。所測試之臨床分離株獲自美國及國外多個地理位置不同之醫院（F〇cusNH OH will be (S)-3-amino-2-(4-(E)-4-(lS,2S)-2-(methyl)cyclopropyl)butan-3-alkynylbenzene Amidoxime _3_methyl butyl methacrylate (4 〇〇 mg, 1 〇 8 mmol, 1.0 eq.) was added to a round bottom flask. To this was added isopropanol (2 mL), the solution was vortexed for 2 minutes, and the flask was cooled in an ice bath, followed by a hydroxylamine solution (1.4 mL, 21.60 mmol, 20 eq.) and the flask was placed at 4. His Majesty 159849.doc • 111 - 201249786 48 hours. The reaction was concentrated (under 〇〇c) to remove isopropanol and then acidified using EtOAc (3 mL). Additional water (15 mL) and ACN (3 mL) were added. The product was purified by reverse phase HPLC (2 "column '5 〇 mL/min, water containing 1% TFA/ACN, equilibrated at 2% B). Using a syringe filter (24 mL) at 10 mL/min The column was loaded with 2% B and then ramped to 5 〇 mL/min over 1 minute. Operates 2% B to 95 in 73 minutes. /. The gradient of B. The desired fractions were combined and frozen and placed on a lyophilizer. The reaction yielded 丨77 mg N-((S)-3-amino-1-(hydroxylamino)_3-methyla· oxetane _2 _ base) _4_((ε)_4·((1,2-trans)2-(hydroxymethyl)cyclopropyl)butene_3_alkynyl)benzamide^22' 卩8. 1^]^]^+1, expected value = 372, 2, observed value = 3 72.2. 4 NMR (DMSO-^6): δ 0.77-0.81 (m, 2H), 1.25 (s, 3H), 1.30 ( s, 3H), 1.34-1.37 (m, 1H), 3.20-3.24 (dd, 1H), 3.37-3.41 (dd, 1H), 4.65-4.67 (d, 1H), 6.43-6.47 (d, 1H), 6.87-6.91 (d, 1H), 7.55-7.57 (d, 2H), 7.84-7.86 (d, 2H), 7.99 (br, 2H), 8.38-8.41 (d, lH), 9.22 (br, 1H), 11.20 (s, 1H). R. Antimicrobial Active Bacteria Screening and Culture Stored at -70 °C in a room temperature at 35 ° C via Miller Hilton Agar (1^1^141·-Hinton agar) (Beckton Dickinson, Franklin Lakes, NJ) The liquid culture bacterial isolate was passaged overnight. The clinical isolates tested were obtained from hospitals with different geographical locations in the United States and abroad (F〇cus

Diagnostics，Herndon, VA and JMI，North Liberty, ΙΑ) ° 品質控制菌株來自美國菌種保存中心（ATCC; Rockville， Md.)。 159849.doc -112- s 201249786 敏感性測試藉由培養液微量稀釋方法根據臨床及實驗室標準研究所 (Clinical and Laboratory Standards Institute ; CLSI)準則測定最低抑制濃度（MIC)。簡言之，將生物體懸浮液調節至 0.5個麥克法蘭標準（McFarland standard)，得到在每毫升 3xlO5與7x105個群落形成單位（CFU)之間之最終接種菌。在無菌陽離子調節米勒希爾頓培養液（Beckton Dickinson) 中製造藥物稀釋液及接種菌。將100 pL接種菌體積添加至含有具有藥物之2倍連續稀釋液之100 μί培養液的孔中。所有接種之微量稀釋盤均在環境空氣中在35°C下培育18至 24小時。在培育之後，將防止可見生長（OD6〇() nm<0.05)之最低藥物濃度記錄為MIC。根據CLSI準則，藉由使用實驗室品質控制菌株及左氧氟沙星（levofloxacin)，即一種具有界定MIC範圍之化合物來監測分析效能。通常，本發明化合物之MIC值為0.03至16 gg/mL。為此，某些代表性化合物之資料示於以下表II中。Diagnostics, Herndon, VA and JMI, North Liberty, ΙΑ) ° Quality control strains from the American Type Culture Collection (ATCC; Rockville, Md.). 159849.doc -112- s 201249786 Sensitivity Test The minimum inhibitory concentration (MIC) was determined by the culture medium microdilution method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Briefly, the biological suspension was adjusted to 0.5 McFarland standard to obtain the final inoculum between 3 x 10 5 and 7 x 105 community forming units (CFU) per ml. Drug dilutions and inoculum were prepared in sterile cation-regulated Miller Hilton B culture (Beckton Dickinson). A 100 pL inoculum volume was added to wells containing 100 μL of culture medium with 2-fold serial dilutions of the drug. All inoculated microdilution plates were incubated for 18 to 24 hours at 35 ° C in ambient air. After incubation, the lowest drug concentration that prevented visible growth (OD6 〇() nm < 0.05) was recorded as MIC. According to the CLSI guidelines, analytical performance is monitored by using laboratory quality control strains and levofloxacin, a compound with a defined MIC range. Generally, the MIC of the compounds of the invention is from 0.03 to 16 gg/mL. To this end, information on certain representative compounds is shown in Table II below.

表II 最低抑制濃度（MIC)Table II Minimum inhibitory concentration (MIC)

化合物編號 APAE001 AEC0001 APAE002 AKPN001 1-1 A A A A 1-2 A A A A 1-3 A A A A 1-4 A A A A 1-5 A A A A 1-6 A A A A 1-7 A A A A 1-8 A A A A 1-9 A A A A 1-10 A A A A 159849.doc •113· 201249786Compound No. APAE001 AEC0001 APAE002 AKPN001 1-1 AAAA 1-2 AAAA 1-3 AAAA 1-4 AAAA 1-5 AAAA 1-6 AAAA 1-7 AAAA 1-8 AAAA 1-9 AAAA 1-10 AAAA 159849.doc • 113· 201249786

1-11 A A A A 1-12 A A A A 1-13 A A A A 1-14 A A A A 1-15 A A A A 1-16 A A A A 1-17 A A A A 1-18 A A A A 1-19 A A A A 1-20 A A A A 1-21 B A A B 1-22 A A A A 1-23 A A A A MIC關鍵字：1-11 AAAA 1-12 AAAA 1-13 AAAA 1-14 AAAA 1-15 AAAA 1-16 AAAA 1-17 AAAA 1-18 AAAA 1-19 AAAA 1-20 AAAA 1-21 BAAB 1-22 AAAA 1- 23 AAAA MIC Keywords:

A=MIC為 2.0 pg/mL 或小於 2.0 pg/mLA=MIC is 2.0 pg/mL or less than 2.0 pg/mL

B=MIC 大於 2·0 pg/mL 至 16.0 pg/mLB=MIC is greater than 2·0 pg/mL to 16.0 pg/mL

C=MIC大於 16.0 gg/mL *AEC0001為大腸桿菌ATCC25922 ; APAE001為綠膿假單胞菌ATCC27853 ； AKPN001為#义3雷伯氏菌 ATCC43816 ; APAE002為表現正常流出活性程度之綠膿假單胞菌之臨床分離株。各.活體内耐受性各化合物經由皮下注射投與小鼠。小鼠在18至28°C及約 5 0%濕度下群居（每籠五隻），且對其餵食標準齧齒動物食物。使其自由取用水及食物。對小鼠投與每公斤體重不大於20mL之皮下劑量。一組三隻小鼠接受皮下注射單次劑量之測試化合物，該化合物係含在由15%坎普的索（Captisol)之20 mM乙酸鹽緩衝液（pH 5)組成之調配物中，每天投與50、100、200、400 或600 mg/kg。在背部肩胛間區域進行皮下注射。將10至C=MIC is greater than 16.0 gg/mL *AEC0001 is Escherichia coli ATCC25922; APAE001 is Pseudomonas aeruginosa ATCC27853; AKPN001 is #3 bacterium 3 ATBE43816; APAE002 is Pseudomonas aeruginosa showing normal efflux activity Clinical isolates. Each in vivo tolerance Each compound was administered to mice via subcutaneous injection. Mice were housed at 18 to 28 ° C and about 50% humidity (five per cage) and fed standard rodent food. Give it free access to water and food. The mice were administered a subcutaneous dose of not more than 20 mL per kilogram of body weight. A group of three mice received a single dose of test compound subcutaneously in a formulation consisting of 15% Campisol 20 mM acetate buffer (pH 5), administered daily. With 50, 100, 200, 400 or 600 mg/kg. Subcutaneous injection is performed in the area between the back and shoulders. Will be 10 to

159849.doc -114- S 201249786 20 mL/kg之體積注射至此區域中《將針平行於皮膚表面插至針尖處於皮下囊袋内之深度。在活塞上施加輕微但穩定之壓力，以排送注射器之内含物。給藥後在以下多個時點進行觀測：30秒至1分鐘、5分鐘、15分鐘、30分鐘、45分鐘、1小時、75分鐘、9〇分鐘、105分鐘、2小時及之後每個小時，直至動物顯示接近基線之恢復徵兆，或直至給藥後4小時（視何者先到達即可）為止。若動物看起來警惕、正常及有反應，則監測小鼠之最小時間範圍為給藥後30分鐘。若動物顯示毒性效應，則對其進行嚴密監測直至其顯示接近基線之恢復徵兆或直至給藥後4小時（視何者先到達即可）為止。動物在給藥之後維持72小時以便臨床觀測，包括監測存活及活動程度。觀測需考慮到中樞神經系統窘迫症之症狀（諸如發作、昏睡、仰臥、不動症、過動症）、神經肌肉異常（諸如運動失調、抽動（twitching)、抽搐（convuisi〇n)、四肢張開、跳或踢）、自主症狀（諸如流涎、流淚、排尿、排便、立毛或斜視）、呼吸窘迫（諸如呼吸困難或呼吸急迫、抑鬱、喘息或喘氣）、定型化行為（諸如反覆咀嚼、繞圈、踱步（pacing)、理毛（grooming)、嗅聞、頭移動或弓背姿態）及行為異常 (諸如逃避行為或濕狗樣抖動）。已知化合物No_aIc(其合成及活性係揭示於國際pct公開案第2008/06676號中；化合物91-12)已顯示當在小鼠中皮下注射時小鼠之最大耐受劑量小於50 mg/kg，而藉由皮下注射需要30 mg/kg之劑量作為在小鼠中針對矽穿伯戌 159849.doc -115- 201249786 磨之靜態劑量。與此已知化合物之不同之處僅在於經經甲基取代之環丙基之本發明化合物1-1具有與N〇_alc抗微生物活性相當之抗微生物活性，但在哺乳動物中具有實質上較佳之耐受性（當皮下注射至小鼠中時，最大耐受劑量為約 200 mg/kg) °159849.doc -114- S 201249786 Volume of 20 mL/kg is injected into this area. Insert the needle parallel to the skin surface to the depth of the needle tip in the subcutaneous pocket. A slight but steady pressure is applied to the piston to dispense the contents of the syringe. Observations were made at various time points after administration: 30 seconds to 1 minute, 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 75 minutes, 9 minutes, 105 minutes, 2 hours, and each hour thereafter. Until the animal shows signs of recovery near the baseline, or up to 4 hours after dosing (as long as it arrives first). If the animal appears vigilant, normal, and responsive, the minimum time frame for monitoring the mice is 30 minutes after dosing. If the animal shows a toxic effect, it is closely monitored until it shows signs of recovery near baseline or up to 4 hours after dosing (when it arrives first). Animals were maintained for 72 hours after dosing for clinical observation, including monitoring for survival and activity. Observations should take into account symptoms of central nervous system obsessive-compulsive disorder (such as seizures, lethargy, supine, inactivity, hyperactivity), neuromuscular abnormalities (such as movement disorders, twitching, convulisi〇n, limb opening) , jumping or kicking), autonomic symptoms (such as salivation, tearing, urination, defecation, standing or strabismus), respiratory distress (such as difficulty breathing or breathing, depression, wheezing or gasping), stereotyped behavior (such as repeated chewing, winding , pacing, grooming, sniffing, head movement or bow-back gestures, and behavioral abnormalities (such as escape behavior or wet dog-like jitter). The compound No_aIc (the synthesis and activity of which is disclosed in International Pct Publication No. 2008/06676; Compound 91-12) has been shown to have a maximum tolerated dose of less than 50 mg/kg when injected subcutaneously in mice. A subcutaneous injection requires a dose of 30 mg/kg as a static dose in the mouse against 矽戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌戌It differs from this known compound only in that the compound 1-1 of the present invention which is substituted with a methyl group-substituted cyclopropyl group has an antimicrobial activity comparable to that of N〇_alc, but has substantially Better tolerance (maximum tolerated dose of about 200 mg/kg when injected subcutaneously into mice) °

表III提供皮下給藥小鼠之觀測結果。耐受性藉由臨床觀察表徵。A類歸為無或有少許諸如以下之中毒症狀之動物· §移動時偶然短暫暫停、輕微呼吸困難或快速恢復 (例如在10分鐘内）之輕微昏睡。8類歸為顯示諸如以下之 -些中毒症狀之動物：移動時暫停較久、具有較長恢復時間（長達1小時）之輕微昏睡，在此期間動物仍能走來走去。 C類歸為具有諸如以下之中度至重度中毒症狀之動物：昏睡、伏臥、伴有斜視及呼吸困難之昏睡、嚴重抽動（跳、踢）或逃避行為。最終，當臨床觀測時間（長達給藥後72小時）内存在任何致死效應（包括需要安樂死之瀕死狀態）時歸為D類。 159849.doc 201249786 表in 活體内耐受性化合物 50 mg/kg 100 mg/1^ 200 mg/kg 400 mg/kg 600 mg/kg No_aIc B B(n=2) C(n=l) C c 1-1 A B c 1-5 A A(n=l) B(n=2) C 1-6 A c c 1-7 A B(n=l) C(n=2) c 1-8 A B(n=l) C(n=2) c 1-9 A C 1-12 A A(n=l) B(n=2) C c 1-13 A B C C(n=l) D(n=2) 1-14 A B(n=2) CCn=l) C(n=2) T.與第二抗菌劑之協同作用在35°c下於環境空氣中經米勒希爾頓瓊脂（Beckton Dickinson，Franklin Lakes，NJ)由摩义浥雲泠代磨（ATCC 43 816)之-70eC冷凍儲備液培養細菌分離株隔夜繼代。藉由培養液微量稀釋方法，根據臨床及實驗室標準研究所 (CLSI)準則測定最低抑制濃度（MIC)。簡言之，將生物體懸浮液調節至0.5個麥克法蘭標準，得到在每毫升3xl〇5與 7xl05個群落形成單位（CFU)之間之最終接種菌。在無菌陽離子調節米勒希爾頓培養液（Beckt〇n Dickins〇n)中製造藥物稀釋液及接種菌。將1〇〇卟接種菌體積添加至含有具有藥物之2倍連續稀釋液之1〇〇叫培養液的孔中。所有接種之微量稀釋盤均在環境空氣中在35t下培育18至24小時。在培育之後，將防止可見生長（Oh⑽nm<〇〇5)之最低藥物 159849.doc -117· 201249786 濃度記錄為MIC。用指定樂劑與N-((S)-3 -胺基-1-(經胺基）-3 -曱基-1-側氧基丁-2-基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁-1，3-二炔基）苯曱醯胺（1-1)之組合進行標準棋盤分析。表IV提供根據標準技術計算之FICI。化合物1-1與萬古黴素、替考拉寧、紅黴素、阿奇黴素、利福平及新生黴素具有活體外協同作用。Table III provides observations of mice administered subcutaneously. Tolerance is characterized by clinical observation. Class A is classified as an animal that has no or a few symptoms of poisoning, such as the following symptoms: § A slight slumber of occasional short pauses during movement, mild dyspnea, or rapid recovery (eg, within 10 minutes). Class 8 is classified as an animal that exhibits some of the symptoms of poisoning: a slight drowsiness that is paused while moving and has a longer recovery time (up to one hour) during which the animal can still walk around. Class C is classified as having animals with moderate to severe symptoms of poisoning such as: slumbering, lying down, lethargy with strabismus and difficulty breathing, severe tics (jumping, kicking) or evasive behavior. Eventually, Class D was classified when there was any lethal effect (including sudden death requiring euthanasia) during clinical observations (up to 72 hours after dosing). 159849.doc 201249786 Table in In vivo Tolerance Compound 50 mg/kg 100 mg/1^ 200 mg/kg 400 mg/kg 600 mg/kg No_aIc BB(n=2) C(n=l) C c 1- 1 AB c 1-5 AA(n=l) B(n=2) C 1-6 A cc 1-7 AB(n=l) C(n=2) c 1-8 AB(n=l) C (n=2) c 1-9 AC 1-12 AA(n=l) B(n=2) C c 1-13 ABCC(n=l) D(n=2) 1-14 AB(n=2 CCn=l) C(n=2) T. Synergistic effect with the second antibacterial agent in the ambient air at 35 ° C by Miller Hilton Agar (Beckton Dickinson, Franklin Lakes, NJ) The bacterial isolates were cultured overnight by the -70eC frozen stock solution of ATCC 43 816. The minimum inhibitory concentration (MIC) was determined by the culture medium microdilution method according to the Clinical and Laboratory Standards Institute (CLSI) guidelines. Briefly, the biological suspension was adjusted to 0.5 McFarland standards to obtain the final inoculum between 3 x 1 〇 5 and 7 x 105 community forming units (CFU) per ml. The drug dilutions and inoculum were prepared in a sterile cation-adjusted Miller's Hilton medium (Beckt〇n Dickins〇n). One ounce of inoculum volume was added to wells containing one sputum culture medium with a 2-fold serial dilution of the drug. All inoculated microdilution plates were incubated in ambient air at 35 t for 18 to 24 hours. After incubation, the lowest drug 159849.doc -117· 201249786 concentration, which prevents visible growth (Oh (10) nm < 〇〇 5), was recorded as MIC. Using a designated agent with N-((S)-3-amino-1-(amino)-3-mercapto-1-yloxybutan-2-yl)-4-(((1,2) Standard chessboard analysis was performed using a combination of -trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (1-1). Table IV provides FICI calculated according to standard techniques. Compound 1-1 has an in vitro synergistic effect with vancomycin, teicoplanin, erythromycin, azithromycin, rifampicin and novobiocin.

表IV FICI 抗生素 FICI(化合物1-1) 萬古黴素 <0.4 替考拉寧 <0.5 紅黴素 0.3 阿奇黴素 0.2 利福平 0.1 新生黴素 0.3 特拉萬星 >2.0 達托黴素 >2.0 利奈唑胺 > 1.5 克林達黴素 0.6 左氧氟沙星 1.0 頭抱 °比普(Ceftobiprole) 1.0 頭孢噻肟 1.0 健大黴素(Gentamycin) 0.5 在中性白血球減少大腿活體内功效模型中檢驗N-((S)-3-胺基-1-(經胺基）-3 -甲基-1-側氧基丁-2 -基）-4-(((1,2 -反）-2_ (羥甲基）環丙基）丁 -1，3-二炔基）苯曱醯胺（1-1)與萬古黴素之活體内協同作用。基本上如Craig及其他人所述運作該 159849.doc -118- 201249786 模型（參見 Gudmundsson等人，「Murine Thigh Infection Model」，Handbook of Animal Models of Infection，M. A. Sande及〇. Zak編；London: Academic Press, 1999，第 137- 144頁）。簡言之’在感染之前’用2劑環磷醯胺使得小鼠中性白血球減少’接著在大腿中，使用1 〇3至丨〇5 CFu尿义克#伯戌磨（ATCC 43 816)之接種菌進行肌肉内感染。在感染後2小時及14小時兩次投與作為陰性對照物之單獨抗生素或媒劑。在實驗期間均維持動物中性白血球減少，以使白血球對感染之作用減至最小，以便由微生物讀出值量測藥物與細菌之活體内相互作用。在感染後24小時，收集大腿，均質化且進行塗鋪，以量測每條大腿存活之CFU數目。感染後2小時亦自動物子群收集大腿，以記錄在即將首次接受抗生素處理（預處理）之前存在之CFU。靜態劑量 (其定義為使24小時時之CFU負荷與感染後〇小時所量測之 CFU負荷一致時的所需劑量）係藉由prizm(Graphpad軟體）中之標準方法，由劑量反應曲線來計算。此等研究之目的為定量評估Li與候選協同劑之組合是否可在此活體内功效模型中，使菌數下降幅度大於各單獨藥劑之降幅總和。對於#义克穿泠代磨ATCC43816感染之治療，萬古黴素顯示與LpxC抑制劑之顯著活體内協同作用。如圖i中所示’每天用220 mg/kg萬古黴素單獨治療感染小鼠時，未使CFU顯著減少。然而，當與化合物w共同給藥時， LpxC抑制劑之靜態劑量顯著降低（圖i及表v)。 159S49.doc -119- 201249786 表v 藥物給藥方案劑量 (mg/kg) 平均 Log1() CFUa 平均Logie CFU相對減少 2小時 26小時 2小時 26小時媒劑每8小時一次 - n.d. 9.95 -3.71 0.00 左氧氟沙星每12小時一次卜10 n.d. 6.48 -0.25 3.46 30 n.d. 4.44 1.79 5.50 100 n.d. 4.07 2.17 5.88 每6小時一次 30 n.d. 10.26 -4.02 -0.31 1-1 100 n.d. 9.51 -3.27 0.44 300 n.d. 7.13 -0.89 2.82 1-1+ 每6小時一次 30+220 n.d. 4.63 1.61 5.32 萬古黴素 100+220 n.d. 4.28 1.95 5.67 300+220 n.d. 3.98 2.26 5.97 萬古黴素+ 1-1 每6小時一次 25+100 n.d· 8.65 -2.41 1.30 75+100 n.d. 8.11 -1.87 1.84 220+100 n.d. 4.99 1.25 4.96 萬古黴素每6小時一次 220 n.d. 10.21 -3.97 -0.26 2小時對照物 n/a - 6.24 n.d. 0.00 3.71 總而言之，此等數據顯示萬古黴素證明驚人之與N_((s)_ 3-胺基·1-(羥胺基）-3-甲基-丨-側氧基丁_2基）4(((12反）_ 2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺之活體内協同作用。應瞭解，本發明之有機化合物可顯示互變異構現象。由於本說明書内之化學結構僅可表示一種可能之互變異構形式，因此應瞭解，本發明涵蓋所繪結構之任何互變異構形式。此外，儘管本文已出於說明之目的顯示及描述了本發明之特定實施例，但當然應瞭解，本發明不限於該等特定實施例’此係因A熟習此項技術者可尤其根據上述教示在不 I59849.doc -120- 201249786 背離本發明之精神及範疇之情況下進行修改之故。因此，本發明除如由隨附申請專利範圍所限制之外不受限制。本說明書所參考之所有美國專利、美國專利申請公開案、美國專利申請案、國外專利、國外專利申請案及非^ 利公開案均以全文引用的方式併入本文中，其引用程度與本說明書不會不一致。【圖式簡單說明】圖1說明在鼠類大腿模型中’菌株atcc 43816 (AKPN001)中化合物w與萬古黴素之總日齊j量及活體内協同作用。 159849.doc 121·Table IV FICI Antibiotic FICI (Compound 1-1) Vancomycin <0.4 Teicoplanin <0.5 Erythromycin 0.3 Azithromycin 0.2 Rifampicin 0.1 Novomycin 0.3 Travancin > 2.0 Daptomycin &gt ;2.0 linezolid> 1.5 clindamycin 0.6 levofloxacin 1.0 head hold Ceftobiprole 1.0 cefotaxime 1.0 gentamicin (Gentamycin) 0.5 in the neutrophil reduction thigh in vivo efficacy model test N -((S)-3-amino-1-(amino)-3-methyl-1-oxobutan-2-yl)-4-(((1,2-trans)-2_ ( In vivo synergy with hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (1-1) and vancomycin. The model 159849.doc -118- 201249786 is basically operated as described by Craig and others (see Gudmundsson et al., "Murine Thigh Infection Model", Handbook of Animal Models of Infection, MA Sande and 〇. Zak; London: Academic Press, 1999, pp. 137-144). Briefly, 'Before infection', use 2 doses of cyclophosphamide to reduce neutrophils in mice'. Then in the thigh, use 1 〇3 to 丨〇5 CFu urinary gram #伯戌磨 (ATCC 43 816) Inoculate bacteria for intramuscular infection. A separate antibiotic or vehicle was administered as a negative control twice at 2 hours and 14 hours after infection. Animal neutropenia was maintained during the experiment to minimize the effects of white blood cells on infection so that the in vivo interaction of the drug with the bacteria was measured by microbial readout values. 24 hours after infection, the thighs were collected, homogenized and spread to measure the number of CFUs survived for each thigh. The thighs were also collected by the automatic object group 2 hours after infection to record the CFU present immediately before the first antibiotic treatment (pretreatment). The static dose (defined as the dose required to match the CFU load at 24 hours to the CFU load measured at the post-infection hour) is calculated from the dose response curve by standard methods in prizm (Graphpad software). . The purpose of these studies was to quantitatively assess whether the combination of Li and the candidate synergist can be used in this in vivo efficacy model to reduce the number of bacteria more than the sum of the individual agents. For the treatment of #伊克穿泠磨磨 ATCC43816 infection, vancomycin showed significant in vivo synergy with LpxC inhibitors. When infected mice were treated with 220 mg/kg vancomycin alone as shown in Figure i, there was no significant reduction in CFU. However, when co-administered with Compound w, the static dose of the LpxC inhibitor was significantly reduced (Figures i and Table v). 159S49.doc -119- 201249786 Table v Drug dosing regimen dose (mg/kg) Average Log1() CFUa Average Logie CFU relative reduction 2 hours 26 hours 2 hours 26 hours vehicle every 8 hours - nd 9.95 -3.71 0.00 Levofloxacin Once every 12 hours, 10 nd 6.48 -0.25 3.46 30 nd 4.44 1.79 5.50 100 nd 4.07 2.17 5.88 Once every 6 hours 30 nd 10.26 -4.02 -0.31 1-1 100 nd 9.51 -3.27 0.44 300 nd 7.13 -0.89 2.82 1-1 + Once every 6 hours 30+220 nd 4.63 1.61 5.32 Vancomycin 100+220 nd 4.28 1.95 5.67 300+220 nd 3.98 2.26 5.97 Vancomycin + 1-1 once every 6 hours 25+100 nd· 8.65 -2.41 1.30 75 +100 nd 8.11 -1.87 1.84 220+100 nd 4.99 1.25 4.96 vancomycin once every 6 hours 220 nd 10.21 -3.97 -0.26 2 hours control n/a - 6.24 nd 0.00 3.71 All in all, these data show vancomycin proof Amazingly with N_((s)-3-amino-1-(hydroxyamino)-3-methyl-indole-sideoxybutan-2-yl)4(((12))-2-(hydroxymethyl) In vivo synergistic action of cyclopropyl)butan-1,3-diynyl)benzamide. It will be appreciated that the organic compounds of the invention may exhibit tautomerism. Since the chemical structures within this specification can only represent one possible tautomeric form, it is to be understood that the present invention encompasses any tautomeric form of the depicted structure. In addition, although the specific embodiments of the present invention have been shown and described herein for purposes of illustration, it is understood that the invention is not limited to the specific embodiments. Modifications are made without departing from the spirit and scope of the invention, in the absence of I59849.doc-120-201249786. Accordingly, the invention is not limited except as limited by the scope of the accompanying claims. All of the U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-disclosed disclosures are hereby incorporated by reference in their entirety in This manual will not be inconsistent. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 illustrates the total amount of the compound w and vancomycin in the strain atcc 43816 (AKPN001) and the in vivo synergistic effect in the murine thigh model. 159849.doc 121·

Claims

201249786 七、申請專利範圍： 1. 一種式I化合物，201249786 VII. Patent application scope: 1. A compound of formula I,

或其立體異構體或醫藥學上可接受之鹽，其中 A為經取代之環烧基，其中個取代基為Cl_ c3—級醇； B不存在，或為_CH=CH_、ac_或未經取代之苯基； C為-CH=Cfc_或未經取代之苯基，其中若B為 -CH-CH- ’ 則 C不會亦為 _CH=CH_ ; R1、R2及R3係獨立地選自氫及經取代或未經取代之^ C3燒基，或 R1及R2連同其所連接之碳原子—起形成未經取代之 C3-C6環烧基，或 R2及R3連同其所連接之碳原子及Q_起形成具有U 8個環原子之經取代或未經取代之雜環，其中該雜環之1至2個環原子係選自N、〇及s ;且 Q為Ο或NR，其中r為氫或未經取代之c^C3烷基。 2.如清求項1之化合物，其申q為nr。 3·如請求項_之化合物’其tRl、R^R3係獨立地選自氫及經取代或未經取代之烷基。 4.如請求項1或2中任一項之化合物，其中Rl、尺2及&3係獨 l59S49.doc 201249786 立地選自氫及未經取代之c〗-c3烷基。 5.如請求項礒2中任一項之化合物，其中咖獨立地為未經取代之cvc3烷基。 6·如請求項即中任-項之化合物，其中A為經經甲基取代之[3-(：6環烷基。 7·如請求項1或2中任一項之化合物，其中3為< =(：_且（：為 -C=C-〇 8.如請求項1之化合物，其中該化合物為： N-((S)-3-胺基-1-(羥胺基）-3-曱基_ι·側氧基丁 _2基）4_ (((1，2-反）-2-(經甲基）環丙基）丁 _1，3_二炔基）笨曱醯胺α- ΐ) ; N-((S)-3-胺基-1-(羥胺基）-3-曱基-1-側氧基丁_2基）4_ (((lR，2R)-2-(經甲基）環丙基）丁-i，3-二炔基）苯甲醯胺（1_ 2); N-((S)_3·胺基-1-(經胺基）-3 -曱基-1-側氧基丁 _2·基）_4_ (((lS，2S)-2-(羥甲基）環丙基）丁-1，3-二炔基）苯甲醯胺（工_ 3) ; N-((S)-3-胺基-1-(羥胺基）-3-甲基-1-側氧基丁 _2_基）_心 (((1，2-順）_2-(經曱基）環丙基）丁 -1，3-二炔基）苯曱醯胺（工_ 4) ； N-((S)-3-胺基-1-(羥胺基）-3-甲基-1-側氧基丁 _2_基）_ 4，-(((1，2-反）-2-(羥甲基）環丙基）乙炔基）聯笨_4-甲醯胺（1_ 5) ; N-((S)-l-(羥胺基）-3-曱基-3-(甲胺基）-1-側氧基丁 _2_ 159849.doc S 201249786 基）-4-(((1,2-反）-2-(羥曱基）環丙基）丁_13_二炔基）苯甲醯胺（1-6); N-((S)-l-(羥胺基）-3-(2-羥基乙胺基）_3_甲基-丨_側氧基丁 -2-基）-4-(((1,2-反）-2-(經曱基）環丙基）丁·13_二炔基）苯甲醯胺（1-7); N-((S)-1 -(羥胺基）-3-(2-羥基乙胺基）·3_曱基_丨_側氧基丁 -2-基）-4-(((1,2-反）-2-(經曱基）環丙基）丁-二炔基）苯甲酿胺（1-8); N-((S)-3-(二甲胺基）-1-(經胺基）_3_甲基_ι_側氧基丁 _2_ 基）-4-((( 1，2-反）-2-(經甲基）環丙基）丁 _1，3_二炔基）苯曱醯胺（1-9); N-((S)-3-羥基-1-(羥胺基）-3-曱基-1-側氧基丁 _2_基）·4_ (((1，2-反）-2-(羥曱基）環丙基）丁-l，3-二炔基）苯甲醯胺（1_ 10); 1<[-((8)-3-胺基-1-(經胺基）-3-曱基-1-側氧基丁_2_基）_4_ (((lS，2R)-2-(羥甲基）-2-曱基環丙基）丁-153_二炔基）苯甲醯胺（1-11); N-((S)-3-胺基-1-(經胺基）-3 -甲基-1-側氧基丁 _2_基） (((1，3-順）-3-(羥甲基）環丁基）丁-1，3-二炔基）苯甲醯胺（1_ 12)； N-((S)-3-胺基-1-(經胺基）-3 -甲基-1-側氧基丁 _2_基）_4_ (((1,3-反）-3-(羥甲基）環丁基）丁-1，3-二炔基）苯甲醯胺（1_ 13)； >1-((8)-3-胺基-1-(經胺基）-3-曱基-1-側氧基丁-2-基）-4- 159849.doc 201249786 (((1，3-反）-3-(羥甲基）環戊基）丁-1，3-二炔基）苯曱酿胺（I- 14) ; N-((S)-3-胺基-1-(羥胺基）-3-甲基-1-側氧基丁-2-基）-4-(((1，4-順）-4-(羥甲基）環己基）丁-1,3-二炔基）苯曱醯胺（I- 15) ； N-(XS)-3-胺基-1-(羥胺基）-3-曱基-1-側氧基丁 -2-基）-4-(((1，4-反）-4-(羥甲基）環己基）丁-1，3-二炔基）苯曱醯胺（I- 16) ； N-((S)-1-(1-胺基環丁基）-2-(羥胺基）-2-側氧基乙基）-4-(((反）-2-(羥曱基）環丙基）丁-1,3-二炔基）苯甲醯胺三氟乙酸鹽（1-17); N-((S)-1-(1-胺基環丁基）-2-(羥胺基）-2-側氧基乙基）-4-(((順）-2-(羥曱基）環丙基）丁-1,3-二炔基）苯曱醯胺三氟乙酸鹽（1-18); N-((S)-1-(1-(二甲胺基）環丁基）-2-(羥胺基）-2-側氧基乙基）_4-(((1，2-反）-2-(羥甲基）環丙基）丁-1，3-二炔基）苯曱醯胺（1-19); N-((S)-2-(經胺基）-l-((S) -嗎琳-3-基）-2 -側氧基乙基）_4-(((1，2-反）-2-(羥甲基）環丙基）丁-1，3-二炔基）苯曱醯胺（I-20)； N-((S)-2-(羥胺基）-l-((S)-4-曱基嗎啉-3-基）-2-側氧基乙基）_4-(((1,2-反）-2-(羥甲基）環丙基）丁-i，3_二炔基）苯甲醯胺（1-21); N-((S)-3-胺基-1-(經胺基）_3_甲基-1·側氧基丁 -2-基）-4- 159849.doc 201249786 ((E)-4-((l，2-反）-:2-(經甲基）環丙基）丁小婦·3_炔基）苯曱醢胺（1-22);或 N-((S)-3-胺基-1-(經胺基）-3 -曱基-1-側氧基丁 _2·基）_4· ((E)-4-((l，2-反）-2-(經甲基）環丙基）丁 _ι_烯_3·炔基）苯甲醯胺（1-23) » 9.如請求項8之化合物，其中該化合物為： N-((<S)-3-胺基-1-(經胺基）-3 -曱基-1-側氧基丁 -2-基）-4-(((I，2·反）_2_(經曱基）環丙基）丁 -1，3-二快基）苯甲酿胺（ι_ 1); N-((S)-3 -胺基-1-(經胺基）-3 -甲基-1-側氧基丁 _2_基）_ 4'-(((1，2-反）-2-(經甲基）環丙基）乙炔基）聯苯_4·曱酿胺（ι_ 5); N-((S)-l-(羥胺基）-3·甲基-3-(曱胺基）4-側氧基丁 _2_ 基）-4-(((1,2-反）-2-(赵甲基）環丙基）丁- i，3_二炔基）苯曱醯胺（1-6); N-((S)-l-(羥胺基）-3-(2-羥基乙胺基）_3_甲基_丨_側氧基丁-2-基）-4-(((1,2-反）-2-(羥甲基）環丙基）丁 ^，3_二炔基）苯甲醯胺（1-7); N-((S)-l-(羥胺基）-3-(2-羥基乙胺基）_3_曱基側氧基丁-2-基）-4-(((1,2-反）-2·(羥甲基）環丙基丨丁一，^二炔基）苯曱醯胺（1-8); N-((S)-3-(二甲胺基）-1-(羥胺基）-3_曱基側氧基丁 _2_ 基）-4-(((1，2-反）-2-(羥曱基）環丙基）丁 4，3_二炔基）笨曱醯胺（1-9); I59849.doc 201249786 N-((S)-3-胺基-W羥胺基）-3-曱基_Μ|ι!氧基丁 _2·基）_4_ (((1，3-順）-3•(經甲基）環T基）丁二炔基）苯甲酿胺⑷ 12)； N-((S)-3·胺基-M羥胺基）·3_甲基-^側氧基丁 _2_基）_4_ (((1，3_反）-3-(經甲基）環丁基）丁二炔基）苯甲醯胺⑷ 13) ;或 N-((s)-3-胺基-M羥胺基）-3_曱基-^側氧基丁_2基）_4_ ((U，3-反(經曱基）環戊基）丁-U·:炔基）苯甲醯胺（Ι· 14) ° 10.如請求項9之化合物，其中該化合物為： Ν-((幻-3-胺基-Μ羥胺基）-3-曱基-I側氧基丁 _2_基）_4· (((1，2-反）-2•(經曱基）環丙基）丁-二炔基）苯曱醢胺（ι· 1); N-((S)-3-胺基-1-(羥胺基）-3-甲基·側氧基丁 _2—基）· 4·-(((1，2·反）-2-(羥甲基）環丙基）乙炔基）聯苯_4甲醯胺（1_ 5); N-((S)-1-(羥胺基）-3 -曱基-3-(甲胺基側氧基丁 _2_ 基）-4-(((1,2-反）-2-(經曱基）環丙基）丁 - i，3_二快基）苯曱醯胺（1-6); N-((S)-l-(經胺基）-3-(2-經基乙胺基）_3_甲基-i_側氧基丁 -2-基）-4-(((1,2-反）-2-(經甲基）環丙基）丁 - i，3_二炔基）苯曱醯胺（1-7); N-((S)-l-(羥胺基）-3-(2-經基乙胺基）-3-甲基側氧基丁 -2-基）-4-(((1，2-反）-2-(經甲基）環丙基）丁 - i，3_二块基） 159849.doc S 201249786 苯甲醯胺（1-8);或 N-((S)-3-(二曱胺基）(經胺基）_3_甲基小側氧基丁-2_ 基）-4-(((1，2-反）-2-(羥甲基）環丙基）丁_1，3_二炔基）苯甲醯胺（1-9)。 11· 一種醫藥組合物，其包含如請求項1至10中任一項之化合物或其醫藥學上可接受之鹽，及醫藥學上可接受之載劑或稀釋劑。 12. —種如請求項1至1〇中任一項之化合物或如請求項醫藥組合物的用途，其用於製造治療患有細菌感染之個體的藥劑。 13. 如請求項11之用途’其中該細菌感染為革蘭氏陰性細菌感染（gram-negative bacterial infection)。 14. 如請求項13之用途’其中該革蘭氏陰性細菌感染為綠膿假單胞菌（PseMc/omowa·? aerwgiwoja)、伯克霍爾德氏菌屬、(五心ha)、腸桿菌科（五wierofeacieriaceae)、弗朗西絲菌屬（FrawchcW/aceae)、沙雷氏菌屬（Serraiza)、變形桿菌屬（Proiew)、克雷伯氏菌屬、腸桿菌屬 (五《ierokcier)、棒檬酸桿菌屬、沙門氏菌 (<Sa/wo«e//a)、普羅威登斯菌屬（ProWA/icia)、耶氏桿菌屬（FemWa)、摩根氏菌屬（Morgawe//a)或大腸桿菌 {Escherichia coli)。 15. 如請求項14之用途，其中該革蘭氏陰性細菌感染為綠膿桿菌綠腹假單胞菌、伯克霍爾德氏菌屬、弗朗西絲菌屬、腸桿菌屬、耶氏桿菌屬或大腸桿菌。 159849.doc 201249786 16. 如請求項15之用途假單胞菌。 17. 如請求項15之用途桿菌。其中該革蘭氏陰性細菌感染為綠膿其中該革蘭氏陰性細菌感染為大腸二抗菌劑及式I化合物Or a stereoisomer or pharmaceutically acceptable salt thereof, wherein A is a substituted cycloalkyl group, wherein one of the substituents is a Cl_c3-ol; B is absent, or is _CH=CH_, ac_ or Unsubstituted phenyl; C is -CH=Cfc_ or unsubstituted phenyl, wherein if B is -CH-CH- ' then C will not be _CH=CH_; R1, R2 and R3 are independent Is selected from hydrogen and substituted or unsubstituted C3 alkyl, or R1 and R2 together with the carbon atom to which they are attached form an unsubstituted C3-C6 cycloalkyl, or R2 and R3 are attached thereto The carbon atom and Q_ form a substituted or unsubstituted heterocyclic ring having U 8 ring atoms, wherein 1 to 2 ring atoms of the hetero ring are selected from N, 〇 and s; and Q is Ο or NR, wherein r is hydrogen or unsubstituted c^C3 alkyl. 2. The compound of claim 1, wherein q is nr. 3. The compound of claim _ wherein tR1, R^R3 are independently selected from hydrogen and substituted or unsubstituted alkyl. 4. The compound of any one of claims 1 or 2, wherein R1, ft 2 and & 3 are independently selected from the group consisting of hydrogen and unsubstituted c-c3 alkyl. 5. The compound of any one of claims 2, wherein the coffee is independently an unsubstituted cvc3 alkyl group. 6. A compound of the above-mentioned claim, wherein A is a methyl-substituted [3-(:6-cycloalkyl). 7. A compound according to any one of claims 1 or 2, wherein 3 is <=(:_ and (: is -C=C-〇8. The compound of claim 1, wherein the compound is: N-((S)-3-amino-1-(hydroxylamino)-3) -曱基_ι·侧oxybutan-2-yl)4_(((1,2-trans)-2-(methyl)cyclopropyl)butyl-1,3-diynyl) Α-ΐ); N-((S)-3-amino-1-(hydroxyamino)-3-indol-1-yloxybutan-2-yl)4_(((lR,2R)-2- (Methyl)cyclopropyl)butyl-i,3-diynyl)benzamide (1_ 2); N-((S)_3·Amino-1-(amino)-3曱-1-yloxybutan-2-yl)_4_(((lS,2S)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide (工_ 3) ; N-((S)-3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)_heart ((1,2-cis)_2 -(via fluorenyl)cyclopropyl)butan-1,3-diynyl)benzamide (Working _ 4); N-((S)-3-Amino-1-(hydroxylamino)-3 -Methyl-1-oxobutan-2-yl)-4,-((1,2-trans)-2-(hydroxymethyl)cyclopropyl)B )) _ 4-carbamamine (1_ 5); N-((S)-l-(hydroxyamino)-3-mercapto-3-(methylamino)-1- oxetane _2 2012 49 159 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 2012 -((S)-l-(hydroxylamino)-3-(2-hydroxyethylamino)_3_methyl-indole_p-oxybutan-2-yl)-4-(((1,2-reverse) )-2-(sulfenyl)cyclopropyl)butyl-13-diynyl) benzamide (1-7); N-((S)-1 -(hydroxylamino)-3-(2- Hydroxyethylamine)·3_mercapto-丨_sideoxybutan-2-yl)-4-(((1,2-trans)-2-(sulfenyl)cyclopropyl)butane-diyne Benzoylamine (1-8); N-((S)-3-(dimethylamino)-1-(amino)_3_methyl_ι_ oxetyl-2-yl) -4-((( 1,2-trans)-2-(methyl)cyclopropyl)butyr-7,3-diynyl)benzamide (1-9); N-((S) 3-hydroxy-1-(hydroxyamino)-3-indolyl-1-yloxybutan-2-yl)·4_(((1,2-trans)-2-(hydroxyindenyl)cyclopropyl D-l,3-diynyl)benzamide (1-10); 1<[-((8)-3-amino-1-(amino)-3-mercapto-1-one side Oxybutan-2-yl)_4_(((lS,2R)-2-(hydroxymethyl)-2-fluorenyl) Propyl)butyl-153-diynyl)benzamide (1-11); N-((S)-3-amino-1-(amino)-3-methyl-1-oxo Ketin-2-yl) (((1,3-cis)-3-(hydroxymethyl)cyclobutyl)butan-1,3-diynyl)benzamide (1-12); N-( (S)-3-amino-1-(amino)-3-methyl-1-oxobutan-2-yl)_4_(((1,3-))-3-(hydroxymethyl) Cyclobutyl)butane-1,3-diynyl)benzamide (1_13); > 1-((8)-3-amino-1-(amino)-3-indenyl -1-Sideoxybutan-2-yl)-4- 159849.doc 201249786 (((1,3-trans)-3-(hydroxymethyl)cyclopentyl)butane-1,3-diynyl) Benzoquinone (I-14); N-((S)-3-amino-1-(hydroxyamino)-3-methyl-1-oxobutan-2-yl)-4-(( (1,4-cis)-4-(hydroxymethyl)cyclohexyl)butyl-1,3-diynyl)phenylhydrazine (I-15); N-(XS)-3-amino-1 -(hydroxyamino)-3-mercapto-1-oxobutan-2-yl)-4-(((1,4-trans)-4-(hydroxymethyl)cyclohexyl)butane-1,3 -diynyl)phenylhydrazine (I-16); N-((S)-1-(1-aminocyclobutyl)-2-(hydroxylamino)-2-yloxyethyl)- 4-(((trans)-2-(hydroxyl)cyclopropyl)butane-1,3-diyne Benzoguanamine trifluoroacetate (1-17); N-((S)-1-(1-aminocyclobutyl)-2-(hydroxylamino)-2-yloxyethyl)- 4-(((cis)-2-(hydroxyindenyl)cyclopropyl)butan-1,3-diynyl)benzoguanamine trifluoroacetate (1-18); N-((S)- 1-(1-(Dimethylamino)cyclobutyl)-2-(hydroxylamino)-2-yloxyethyl)-4-((1,2-trans)-2-(hydroxymethyl) Cyclopropyl)butan-1,3-diynyl)benzamide (1-19); N-((S)-2-(amino)-l-((S)-Merlin-3 -yl)-2-oxoethyl) 4-((1,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-1,3-diynyl)benzamide I-20); N-((S)-2-(hydroxylamino)-l-((S)-4-indolylmorpholin-3-yl)-2-yloxyethyl)_4-(( (1,2-trans)-2-(hydroxymethyl)cyclopropyl)butan-i,3-diynyl)benzamide (1-21); N-((S)-3-amino -1-(amino group)_3_methyl-1·sideoxybutan-2-yl)-4- 159849.doc 201249786 ((E)-4-((l,2-re)-:2- (Methyl)cyclopropyl)butanyl-3-ynyl)benzamide (1-22); or N-((S)-3-amino-1-(amino)-3 - mercapto-1-yloxybutan-2-yl)_4· ((E)-4-((l,2-trans)-2-( ))))))))))))))))))))))))))))))) 3-amino-1-(amino-amino)-3-indolyl-1-yloxybutan-2-yl)-4-(((I,2·trans)_2_(fluorenyl)cyclopropyl) )-1,3-1,3-diyl)benzamide (ι-1); N-((S)-3-amino-1-(amino)-3-methyl-1-oxyl _2_2_yl)_ 4'-(((1,2-trans)-2-(methyl)cyclopropyl)ethynyl)biphenyl_4·曱-amine (ι_ 5); N-( (S)-l-(hydroxylamino)-3.methyl-3-(decylamino)4-oxobutan-2-yl)-4-(((1,2-trans)-2-(radiomethyl) Cyclopropyl)butan-i,3-diynyl)phenylhydrazine (1-6); N-((S)-l-(hydroxylamino)-3-(2-hydroxyethylamino)_3 _Methyl_丨_Phenyloxybutan-2-yl)-4-(((1,2-trans)-2-(hydroxymethyl)cyclopropyl)butane, 3-diynyl)benzamide Indoleamine (1-7); N-((S)-l-(hydroxylamino)-3-(2-hydroxyethylamino)_3_indolyloxybutan-2-yl)-4-(( (1,2-trans)-2·(hydroxymethyl)cyclopropyl hydrazin-, di-diynyl) benzoguanamine (1-8); N-((S)-3-(dimethylamine) Base-1-(hydroxylamino)-3_fluorenyl Oxybutan-2-yl)-4-(((1,2-trans)-2-(hydroxyindenyl)cyclopropyl)butane 4,3-diynyl) azainamine (1-9); I59849.doc 201249786 N-((S)-3-Amino-Whydroxylamino)-3-indolyl_Μ|ι!oxybutan-2-yl)_4_ (((1,3-cis)-3 • (via methyl) ring T-based) butadiynyl) benzoic acid (4) 12); N-((S)-3·amino-Mhydroxylamino)·3_methyl-^-oxylated _2_yl)_4_(((1,3_trans)-3-(methyl)cyclobutyl)butadiynyl)benzamide (4) 13) ; or N-((s)-3- Amino-M-hydroxylamino)-3_indolyl-^-oxybutylbutanyl)_4_((U,3-trans(sulfenyl)cyclopentyl)butan-U·:alkynyl)benzimidazole Amine (Ι·14) ° 10. The compound of claim 9, wherein the compound is: Ν-((Phase-3-amino-hydroxylamino)-3-indolyl-I-side oxybutane_2_ Base)_4·(((1,2-trans)-2•(sulfenyl)cyclopropyl)butan-diynyl)benzamide (ι·1); N-((S)-3- Amino-1-(hydroxyamino)-3-methyl-t-oxybutan-2-yl)·4·-((1,2·trans)-2-(hydroxymethyl)cyclopropyl)acetylene Biphenyl) 4-carbamamine (1_ 5); N-((S)-1-(hydroxylamino)-3-mercapto-3-(A) Alkyl oxybutyryl-2-yl)-4-(((1,2-trans)-2-(sulfenyl)cyclopropyl)butan-i,3-di-furyl)benzamide (1- 6); N-((S)-l-(amino-)-3-(2-aminoethylamino)_3_methyl-i-sideoxybutan-2-yl)-4-(( (1,2-trans)-2-(methyl)cyclopropyl)butan-i,3-diynyl) benzoguanamine (1-7); N-((S)-l-(hydroxylamine) 3-(2-ethylethylamino)-3-methyl-oxobutan-2-yl)-4-(((1,2-trans)-2-(methyl)cyclopropane )) - i,3_二块基) 159849.doc S 201249786 Benzamide (1-8); or N-((S)-3-(diodino) (amino group)_3_ Methyl-small-oxybutyl-2-yl)-4-(((1,2-trans)-2-(hydroxymethyl)cyclopropyl)butyr-7,3-diynyl)benzamide ( 1-9). A pharmaceutical composition comprising the compound of any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent. 12. Use of a compound according to any one of claims 1 to 1 or a pharmaceutical composition according to claim 1 for the manufacture of a medicament for treating a subject having a bacterial infection. 13. The use of claim 11 wherein the bacterial infection is a gram-negative bacterial infection. 14. For the use of claim 13 'where the Gram-negative bacterial infection is Pseudomonas aeruginosa (PseMc/omowa·? aerwgiwoja), Burkholderia, (five-heart ha), Enterobacter Branch (five wierofeacieriaceae), Frawchc W/aceae, Serraiza, Proiew, Klebsiella, Enterobacter (five "ierokcier", citrate Bacillus, Salmonella (<Sa/wo«e//a), ProWA/icia, FemWa, Morgawe//a or Escherichia coli {Escherichia coli). 15. The use of claim 14, wherein the Gram-negative bacterial infection is Pseudomonas aeruginosa, Pseudomonas cepacia, Francis, Enterobacter, Yersinia or E. coli. 159849.doc 201249786 16. Use of claim 15 Pseudomonas. 17. Use of bacillus as claimed in item 15. Wherein the Gram-negative bacterial infection is P. aeruginia, wherein the Gram-negative bacterial infection is a large intestine antibacterial agent and a compound of formula I

或其立體異構體或醫藥學上可接受之鹽，其中 18· —種醫藥組合物，其包含第 A為經取代之C3_C6環院基，其中至卜個取代基為^ C3—級醇； B不存在，終，.…。或未經取代之苯基； C為-CH=CH_、《_或未經取代之苯基其中若b為 -CH=CH- ’ 則 C不會亦為 _CH=CH-; R、R2及R3係獨立地選自氫及經取代或未經取代之 C3燒基，或 R丨及R2連同其所連接之碳原子一起形成未經取代之匸3-〇：6環烧基，或 R2及R3連同其所連接之碳原子及Q—起形成具有5至 8個環原子之經取代或未經取代之雜環，彡中該雜環之1至2個環原子係選自n、〇及s ;且义 Q為0或NR，其中R為氫或未經取代之C1_C3烷基。 19.如請求項丨8之醫藥組合物，其中該抗菌劑為萬古 I59849.doc 201249786 (vancomycin)或利福平（rifampin)。 20.如請求項1 8之醫藥組合物’其中該組合證明活體内協同作用。 21. —種如請求項1至10中任一項之化合物或如請求項^之醫藥組合物的用途’其用於製造抑制革蘭氏陰性細菌中之去乙醯基酶的藥劑。 22. 如請求項21之用途，其中該革蘭氏陰性細菌為綠膿假單胞菌、伯克霍爾德氏菌屬、腸桿菌科、弗朗西絲菌屬、沙雷氏菌屬、變形桿菌屬、克雷伯氏菌屬、腸桿菌屬、檸檬酸桿菌屬、沙門氏菌、普羅咸登㈣屬、屬、摩根氏菌屬或大腸桿菌。干 23. 一種如請求項^中任-項之化合物或如請醫藥組合物的用途，其用於製造項之 LPXC的藥劑。㈣㈣氏陰性細菌中 159849.docOr a stereoisomer or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition comprises a substituted C3_C6 ring-based group, wherein the substituent is a C-C 3 -alcohol; B does not exist, at the end, .... Or unsubstituted phenyl; C is -CH=CH_, "_ or unsubstituted phenyl wherein if b is -CH=CH-' then C will not be _CH=CH-; R, R2 and R3 is independently selected from hydrogen and substituted or unsubstituted C3 alkyl, or R and R2 together with the carbon atom to which they are attached form an unsubstituted 匸3-〇:6-ring group, or R2 and R3, together with the carbon atom to which it is attached, and Q, form a substituted or unsubstituted heterocyclic ring having 5 to 8 ring atoms, wherein 1 to 2 ring atoms of the hetero ring are selected from n, fluorene and And wherein Q is 0 or NR, wherein R is hydrogen or an unsubstituted C1_C3 alkyl group. 19. The pharmaceutical composition according to claim 8, wherein the antibacterial agent is Ivana I59849.doc 201249786 (vancomycin) or rifampin. 20. The pharmaceutical composition of claim 18 wherein the combination demonstrates in vivo synergy. 21. Use of a compound according to any one of claims 1 to 10 or a pharmaceutical composition according to claim 1 for use in the manufacture of a medicament for inhibiting deacetylase in Gram-negative bacteria. 22. The use of claim 21, wherein the Gram-negative bacterium is Pseudomonas aeruginosa, Burkholderia, Enterobacteriaceae, Francis, Serratia, Proteus , Klebsiella, Enterobacter, Citrobacter, Salmonella, Proton (four) genus, genus, Morganella or Escherichia coli. Dry 23. A compound of any of the claims or a pharmaceutical composition for use in the manufacture of a LPXC agent. (d) (four) negative bacteria 159849.doc