TWI695841B

TWI695841B - Prodrugs for the treatment of cancer

Info

Publication number: TWI695841B
Application number: TW108134937A
Authority: TW
Inventors: 理查畢席爾; 安德斯尹爾洛斯; 柏裘恩卡拉森; 佛德瑞克奧柏格
Original assignee: 瑞典商米迪維艾克提伯拉公司
Priority date: 2015-06-22
Filing date: 2015-08-26
Publication date: 2020-06-11
Also published as: TW201700487A; TW201945378A; TWI687431B

Abstract

The invention provides compounds of the formula:

R¹ is OR¹¹ , or NR⁵ R^5’ ;R² is H or F;R⁵ is H, C₁ -C₆ alkyl, OH, C(=O)R⁶ , O(C=O)R⁶ or O(C=O)OR⁶ ;R^5´ is H or C₁ -C₆ alkyl;R⁶ is C₁ -C₆ alkyl or C₃ -C₇ cycloalkyl;R¹³ is H, phenyl, pyridyl, benzyl, indolyl or naphthyl wherein the phenyl, pyridyl, benzyl, indolyl and naphthyl is optionally substituted with 1, 2 or 3 R²² ; and the other variables are as defined in the claims, which are of use in the treatment of cancer, and related aspects.

Description

Pre-medication for cancer

本發明係關於曲沙他濱(troxacitabine)之磷前藥以及其衍生物，其用於治療癌症，尤其肝癌(諸如肝細胞癌(HCC))及繼發性肝癌。本發明進一步係關於包含此等化合物之組合物及組合，以及其用於治療癌症，尤其肝癌(諸如HCC)之方法。The present invention relates to phosphorus prodrugs of troxacitabine and derivatives thereof, which are used to treat cancer, especially liver cancer (such as hepatocellular carcinoma (HCC)) and secondary liver cancer. The present invention further relates to compositions and combinations comprising these compounds, and methods for treating cancer, especially liver cancer, such as HCC.

原發性肝癌為全球第六大最常見癌症且為癌症死亡之第二大起因。佔所有原發性惡性肝癌之約85%且發病率升高之最常見肝癌為肝細胞癌(HCC)，其係藉由肝細胞變成惡性而形成。另一種由肝細胞形成之癌症為肝母細胞瘤，一種多發於兒童之罕見惡性腫瘤，並佔兒童所有癌症之約1%且佔15歲以下所有原發性肝癌之79%。繼發性肝癌或肝轉移為一種在體內某處開始並隨後擴散到肝臟之癌症。繼發性肝癌之實例包括多種常見形式之癌症，諸如結腸癌、直腸癌、肺癌以及乳癌。肝癌亦可由肝臟內之其他結構形成，諸如膽管、血管及免疫細胞。膽管癌(肝膽管型肝癌及膽管細胞囊腺癌)佔原發性肝癌之約6%。雖然對於早期HCC，手術切除及肝移植為可能具治癒性的療法，但20%以上的患者最終將復發或遇到其他問題，並且大多數HCC診斷出時所處之階段對於此等治療而言過於晚期。區域療法(諸如射頻消融)與大於60%之響應速率相關，但是其僅適於一定比例的患者且並不一直具有治癒性。迄今使用之化療對HCC有效性很小且響應速率不超過25%。目前，索拉非尼(sorafenib)為市面上用於治療晚期或不可切除之HCC之唯一有效藥物，因此，對HCC之其他治療以降低復發率並增加總存活率之需求很大。發現多種核苷類似物具有抗癌活性並且其構成廣泛用於治療癌症患者之一類主要之化療劑。稱為抗代謝物之此類藥劑包括具有細胞毒活性之多種嘧啶及嘌呤核苷衍生物。細胞核苷激酶將核苷磷酸化為其相應之5'-單磷酸鹽，5'-單磷酸鹽經進一步轉化為其二磷酸鹽並隨後轉化為醫藥活性三磷酸鹽。已知一些核苷因為不能被激酶有效地磷酸化或根本不為激酶之受質而活性弱。在磷酸化序列中，核苷類似物之第一磷酸化為限速性的，但第二及第三磷酸化對核苷修飾不太敏感。單磷酸核苷(核苷酸)本身一般在血液中不穩定並表現出不良的膜滲透性因此不適於用作藥物。由於核苷及核苷類似物之三磷酸鹽之高不穩定性及不良細胞滲透性，所以其亦不被視為可能的藥物候選。曲沙他濱(β-L-二氧雜環戊烷胞苷)為一種具有非天然L-組態之細胞毒性脫氧胞苷類似物，其在活體外及活體內已顯示針對實體及造血系統惡性腫瘤之廣泛活性。特定言之，已觀察到針對人類癌細胞系及肝細胞源、前列腺源及腎源之異種移植物之突出活性(Cancer Res., 55, 3008-3011, 1995)。曲沙他濱已證實導致通常負責核苷之第一磷酸化步驟之激酶脫氧胞苷激酶(dCK)之突變，導致沒有或極少量曲沙他濱單磷酸鹽產生，進而產生抗性。在2008年，曲沙他濱進入急性髓性白血病適應症之III期臨牀試驗，但未進展至註冊。中斷的曲沙他濱II期試驗包括乳癌、結直腸癌、胰腺癌、黑素瘤、NSCLC、腎瘤、前列腺瘤及卵巢瘤。曲沙他濱通常以靜脈內輸注之形式投與，進而使多種組織暴露於藥物，這與癌症的位置無關。已證實曲沙他濱儘管其為親水性的但仍藉由被動擴散輸送到細胞中，但與其他(經載體輸送的核苷)相比，僅為極緩慢地積聚在癌細胞中。在WO2008/030373中，揭示了在胞嘧啶鹼基部分上攜載前藥基團的曲沙他濱衍生物並評價了前藥之親脂性與其抗腫瘤活性之間的關係。該專利指出需要鹼基修飾以避免脂酶難以經5'-OH修飾。 D-核苷之5'羥基官能基之胺基磷酸酯前藥已成功用於抗病毒藥物，諸如用於治療HCV感染之索菲布韋(sofosbuvir)。對索菲布韋前藥去遮蔽以在細胞內暴露單磷酸鹽為一個複雜的多步驟過程，其涉及呈特定序列之若干水解酶。胺基磷酸酯前藥很少成功地用於癌症核苷。Nucana正開發Acelerin(Nuc-1031)，一種用於治療胰腺癌之D-核苷吉西他濱(gemcitabine)之胺基磷酸酯前藥(對於結構：參見WO2005012327中第71頁)。然而，即使胺基磷酸酯被認為可提高化合物之親脂性及細胞滲透性，但Acelarin前藥必須仍以靜脈內輸注形式投與，因此將多種健康組織暴露於細胞毒性代謝物。甚至很少利用L-核苷(例如曲沙他濱)之單磷酸鹽前藥。WO2008048128揭示了少量曲沙他濱單磷酸鹽前藥，包括實例14之化合物：

在WO2008048128說明書或學術文獻中其他處中，沒有揭示任何化合物之癌症或其他生物活性。沒有關於該種前藥進入臨牀試驗之報導。然而，WO2008048128之發明人公佈了以下各物之大致類似之前藥：D-核苷吉西他濱(Baraniak等人Biorg Med Chem 2014 2133-2040)，其中前藥方法似乎在某些組織中起作用，以及D-核苷曡氮胸苷(Kulic等人Antivir Chem Chemother 2011 21(3) 143-150)，其中前藥比相應的母體核苷之效力低2-20倍。Kulic推測曡氮胸苷前藥傾向於先去磷酸化為核苷隨後僅磷酸化為活性三磷酸鹽物質。鑒於前藥方法作用於吉西他濱(gemcitabine)(其藉由其經取代之2'官能基而類似於RNA)並不作用於曡氮胸苷(其為2'-脫氧，從而類似於DNA)中，假設WO2008048128中曲沙他濱之前藥與曡氮胸苷前藥一樣可能無活性。 Balzarini等人Biochem Biophys Res Comm 225, 363-369 (1996)描述了CF 1109之HIV及HBV活性，CF 1109為一種L-核苷拉米夫定(lamivudine)/3TC之胺基磷酸酯前藥，其具有以下結構式：

Balzarini指出該胺基磷酸酯前藥對抗HIV比其母體核苷3TC的活性低~250倍，但前藥「在Hep G2.2.15細胞中對抗HBV幾乎等效」。換言之，此大(胺基磷酸酯甲酯)基團之添加不改善在肝細胞株中之抗病毒效力。Balzarini未分析前藥在磷酸化為活性三磷酸鹽前是否代謝為3TC。本發明者提供了曲沙他濱之磷前藥，特定言之靶向肝之前藥，諸如胺基磷酸酯，其適於經口投與。此等前藥因相比於曲沙他濱自身之親脂性增加而具有改良之細胞滲透性，並因繞過限速性第一磷酸化步驟而更有效地形成活性三磷酸鹽。此外，本發明化合物在肝臟中主要代謝為活性三磷酸鹽，從而在靶器官中提供高濃度之活性化合物並同時將在其他器官中之因毒性所致之副作用保持在最低。Primary liver cancer is the sixth most common cancer in the world and the second leading cause of cancer death. The most common liver cancer that accounts for about 85% of all primary malignant liver cancers and has an increased incidence is hepatocellular carcinoma (HCC), which is formed by liver cells becoming malignant. Another type of cancer formed by hepatocytes is hepatoblastoma, a rare malignant tumor that occurs frequently in children and accounts for about 1% of all cancers in children and 79% of all primary liver cancers under the age of 15. Secondary liver cancer or liver metastasis is a type of cancer that starts somewhere in the body and then spreads to the liver. Examples of secondary liver cancer include many common forms of cancer, such as colon cancer, rectal cancer, lung cancer, and breast cancer. Liver cancer can also be formed by other structures in the liver, such as bile ducts, blood vessels, and immune cells. Cholangiocarcinoma (hepatobiliary hepatocellular carcinoma and cholangiocarcinoma) accounts for about 6% of primary liver cancer. Although for early HCC, surgical resection and liver transplantation are potentially curative therapies, more than 20% of patients will eventually relapse or encounter other problems, and most HCCs are diagnosed at the stage for these treatments Too late. Regional therapies (such as radiofrequency ablation) are associated with response rates greater than 60%, but they are only suitable for a certain percentage of patients and are not always curative. The chemotherapy used so far has little effect on HCC and the response rate does not exceed 25%. Currently, sorafenib is the only effective drug on the market for the treatment of advanced or unresectable HCC. Therefore, there is a great demand for other treatments of HCC to reduce the relapse rate and increase the overall survival rate. Various nucleoside analogs have been found to have anti-cancer activity and they constitute one of the major chemotherapeutic agents widely used in the treatment of cancer patients. Such agents called antimetabolites include various pyrimidine and purine nucleoside derivatives with cytotoxic activity. The nucleoside kinase phosphorylates the nucleoside to its corresponding 5'-monophosphate, which is further converted into its diphosphate and then into the pharmaceutically active triphosphate. It is known that some nucleosides have weak activity because they cannot be effectively phosphorylated by kinases or are not substrates of kinases at all. In the phosphorylation sequence, the first phosphorylation of the nucleoside analog is rate-limiting, but the second and third phosphorylation are less sensitive to nucleoside modification. Nucleoside monophosphate (nucleotide) itself is generally unstable in blood and exhibits poor membrane permeability and is therefore not suitable for use as a drug. Due to the high instability and poor cell permeability of triphosphates of nucleosides and nucleoside analogs, they are not considered as possible drug candidates. Trisatabine (β-L-dioxolane cytidine) is a cytotoxic deoxycytidine analog with unnatural L-configuration, which has been shown to target entities and hematopoietic systems in vitro and in vivo Extensive activity of malignant tumors. In particular, outstanding activity against human cancer cell lines and hepatocyte-derived, prostate-derived and kidney-derived xenografts has been observed (Cancer Res., 55, 3008-3011, 1995). Triscitabine has been shown to cause mutations in the kinase deoxycytidine kinase (dCK), which is usually responsible for the first phosphorylation step of the nucleoside, resulting in no or minimal production of trisatabine monophosphate, which in turn leads to resistance. In 2008, trisatabine entered the phase III clinical trial for indications of acute myeloid leukemia, but did not progress to registration. The discontinued phase 2 trial of tricabine included breast cancer, colorectal cancer, pancreatic cancer, melanoma, NSCLC, renal tumor, prostate tumor, and ovarian tumor. Trisatabine is usually administered as an intravenous infusion, which exposes multiple tissues to drugs, regardless of the location of the cancer. Trisapitabine has been confirmed to be transported into cells by passive diffusion, although it is hydrophilic, but it accumulates in cancer cells only very slowly compared to others (nucleosides delivered via a carrier). In WO2008/030373, a trastabine derivative carrying a prodrug group on a cytosine base portion is disclosed and the relationship between the lipophilicity of the prodrug and its antitumor activity is evaluated. The patent states that base modification is needed to avoid the difficulty of lipase modification by 5'-OH. Aminophosphate prodrugs of the 5'hydroxyl functional group of D-nucleosides have been successfully used in antiviral drugs, such as sofosbuvir for the treatment of HCV infection. Demasking the prodrugs of Sofibuvir to expose the monophosphate in the cell is a complex multi-step process that involves several hydrolases in a specific sequence. Aminophosphate prodrugs have rarely been successfully used in cancer nucleosides. Nucana is developing Acelerin (Nuc-1031), an aminophosphate prodrug of D-nucleoside gemcitabine for the treatment of pancreatic cancer (for structure: see page 71 in WO2005012327). However, even if aminophosphates are believed to increase the lipophilicity and cell permeability of the compound, Acelarin prodrug must still be administered as an intravenous infusion, thus exposing a variety of healthy tissues to cytotoxic metabolites. The monophosphate prodrugs of L-nucleosides (e.g. trisatabine) are even rarely used. WO2008048128 discloses a small amount of trisatabine monophosphate prodrug, including the compound of Example 14:

In the WO2008048128 specification or elsewhere in the academic literature, no cancer or other biological activity of any compound is disclosed. There are no reports of this prodrug entering clinical trials. However, the inventors of WO2008048128 have published prodrugs that are roughly similar to the following: D-nucleoside gemcitabine (Baraniak et al. Biorg Med Chem 2014 2133-2040), where the prodrug approach seems to work in certain tissues, and D -Nucleoside ribothymidine (Kulic et al. Antivir Chem Chemother 2011 21(3) 143-150), where the prodrug is 2-20 times less potent than the corresponding parent nucleoside. Kulic speculates that azathiothymidine prodrugs tend to dephosphorylate to nucleosides first and then phosphorylate only to active triphosphates. Given that the prodrug method acts on gemcitabine (which is similar to RNA by its substituted 2'functional group) and does not act on azathiothymidine (which is 2'-deoxy, thus similar to DNA), It is assumed that the prodrug of Qusartabine in WO2008048128 may be as inactive as the prodrug of azathymidine. Balzarini et al. Biochem Biophys Res Comm 225, 363-369 (1996) describe the HIV and HBV activities of CF 1109, which is an L-nucleoside lamivudine/3TC aminophosphate prodrug, It has the following structural formula:

Balzarini pointed out that the aminophosphate prodrug is about 250 times less active against HIV than its parent nucleoside 3TC, but the prodrug is "almost equivalent to fighting HBV in Hep G2.2.15 cells." In other words, the addition of this large (methyl phosphoramidate) group does not improve the antiviral efficacy in liver cell lines. Balzarini did not analyze whether the prodrug was metabolized to 3TC before being phosphorylated to active triphosphate. The inventors provided phosphorus prodrugs of trichatabine, specifically targeting liver prodrugs, such as aminophosphates, which are suitable for oral administration. These prodrugs have improved cell permeability due to increased lipophilicity compared to tricatabine itself, and more efficiently form active triphosphates by bypassing the rate-limiting first phosphorylation step. In addition, the compounds of the present invention are mainly metabolized in the liver to active triphosphates, thereby providing high concentrations of active compounds in target organs while keeping side effects due to toxicity in other organs to a minimum.

在一態樣中，本發明提供由式(I)表示之化合物：

R¹ 為OR¹¹ 、或NR⁵ R^5' ； R² 為H或F； R⁵ 為H、C₁ -C₆ 烷基、OH、C(=O)R⁶ 、OC(=O)R⁶ 或OC(=O)OR⁶ ； R^5' 為H或C₁ -C₆ 烷基； R⁶ 為C₁ -C₂₂ 烷基或C₃ -C₇ 環烷基； R¹¹ 為H或C₁ -C₆ 烷基； R¹³ 為H、苯基、吡啶基、苄基、吲哚基或萘基，其中該苯基、吡啶基、苄基、吲哚基及萘基視情況經1、2或3個R²² 取代； R¹⁵ 為H、C₁ -C₆ 烷基、C₃ -C₇ 環烷基、C₃ -C₇ 環烷基C₁ -C₃ 烷基、苯基、苄基或吲哚基； R^15' 為H或C₁ -C₆ 烷基；或 R¹⁵ 與R^15' 連同其所連接的碳原子一起形成C₃ -C₇ 伸環烷基，其中各C₁ -C₆ 烷基視情況經選自鹵基、OR¹⁸ 及SR¹⁸ 的基團取代，且各C₃ -C₇ 環烷基、C₃ -C₇ 伸環烷基、苯基及苄基視情況經一或兩個獨立地選自C₁ -C₃ 烷基、鹵基及OR¹⁸ 的基團取代； R¹⁶ 為H、C₁ -C₁₀ 烷基、C₂ -C₁₀ 烯基、C₃ -C₇ 環烷基、C₃ -C₇ 環烷基C₁ -C₃ 烷基、苄基或苯基，其中任一者視情況經1、2或3個各自獨立地選自鹵基、OR¹⁸ 及N(R¹⁸ )₂ 的基團取代；各R¹⁸ 獨立地為H、C₁ -C₆ 烷基、C₁ -C₆ 鹵烷基或C₃ -C₇ 環烷基；各R²² 獨立地選自鹵基、C₁ -C₆ 烷基、C₂ -C₆ 烯基、C₁ -C₆ 鹵烷基、C₁ -C₆ 烷氧基、C₁ -C₆ 鹵烷氧基、苯基、羥基C₁ -C₆ 烷基、C₃ -C₆ 環烷基、C₁ -C₆ 烷基羰基、C₃ -C₆ 環烷基羰基、羧基C₁ -C₆ 烷基、羥基、胺基CN以及NO₂ ，或連接至相鄰環碳原子的任意兩個R²² 基團可組合形成-O-(CR²³ R^23' )_1-6 -O-； R²³ 及R^23' 獨立地為H或C₁ -C₃ 烷基；或其醫藥學上可接受的鹽及/或溶劑合物。在一個實施例中，本發明提供由式I表示之化合物：

其中： R¹ 為OR¹¹ 或NR⁵ R^5' ； R² 為H或F； R⁵ 為H、C₁ -C₆ 烷基、OH、C(=O)R⁶ 、OC(=O)R⁶ 或OC(=O)OR⁶ ； R^5' 為H或C₁ -C₆ 烷基； R⁶ 為C₁ -C₂₂ 烷基或C₃ -C₇ 環烷基； R¹¹ 為H或C₁ -C₆ 烷基； R¹³ 為H、苯基、吡啶基、苄基、吲哚基或萘基，其中該苯基、吡啶基、苄基、吲哚基及萘基視情況經1、2或3個R²² 取代； R¹⁵ 為H、C₁ -C₆ 烷基、C₃ -C₇ 環烷基、C₃ -C₇ 環烷基C₁ -C₃ 烷基、苯基、苄基或吲哚基； R^15' 為H或C₁ -C₆ 烷基；或 R¹⁵ 與R^15' 連同其所連接的碳原子一起形成C₃ -C₇ 伸環烷基，其中各C₁ -C₆ 烷基視情況經選自鹵基、OR¹⁸ 及SR¹⁸ 的基團取代，且各C₃ -C₇ 環烷基、C₃ -C₇ 伸環烷基、苯基及苄基視情況經一或兩個獨立地選自C₁ -C₃ 烷基、鹵基及OR¹⁸ 的基團取代； R¹⁶ 為H、C₁ -C₁₀ 烷基、C₂ -C₁₀ 烯基、C₃ -C₇ 環烷基、C₃ -C₇ 環烷基C₁ -C₃ 烷基、苄基或苯基，其中任一者視情況經1、2或3個各自獨立地選自鹵素、OR¹⁸ 及N(R¹⁸ )₂ 的基團取代；各R¹⁸ 獨立地為H、C₁ -C₆ 烷基、C₁ -C₆ 鹵烷基或C₃ -C₇ 環烷基；各R²² 獨立地選自鹵基、C₁ -C₆ 烷基、C₂ -C₆ 烯基、C₁ -C₆ 鹵烷基、C₁ -C₆ 烷氧基、C₁ -C₆ 鹵烷氧基、苯基、羥基C₁ -C₆ 烷基、C₃ -C₆ 環烷基、C₁ -C₆ 烷基羰基、C₃ -C₆ 環烷基羰基、羧基C₁ -C₆ 烷基、羥基、胺基CN、NO₂ 以及三甲基矽烷基，或連接至相鄰環碳原子之任意兩個R²² 基團可組合形成-O-(CR²³ R^23' )_1-6 -O-； R²³ 及R^23' 獨立地為H或C₁ -C₃ 烷基；或其醫藥學上可接受的鹽及/或溶劑合物。式(I)化合物可視情況以醫藥學上可接受之鹽及/或溶劑合物形式提供。在一個實施例中，本發明之化合物係以醫藥學上可接受之鹽提供。在第二實施例中，本發明之化合物係以醫藥學上可接受之溶劑合物形式提供。在第三實施例中，本發明之化合物係以其游離形式提供。在本發明之代表性實施例中，R¹ 為NR⁵ R^5' ，諸如NH₂ 或NHC(=O)C₁ -C₆ 烷基。 R² 一般為H。在較佳實施例中，R¹ 為NH₂ 且R² 為H。在替代性實施例中，R¹ 為NH₂ 且R² 為F。典型地，在式(I)化合物中，部分-NHC(R¹⁵ )(R^15' )-C(=O)OR¹⁶ 形成胺基酸酯殘基，包括天然及非天然胺基酸殘基。特別關注其中R^15' 為氫且R¹⁵ 為甲基、異丙基、異丁基或苄基之胺基酸殘基。在一典型組態中，R^15' 為H且R¹⁵ 為C₁ -C₃ 烷基，諸如甲基、乙基、丙基、異丙基。在其中R^15' 為氫且R¹⁵ 不為氫之化合物中，不對稱碳原子之組態一般為L-胺基酸之組態，因此提供具有式(Ia)中所示之立體化學之化合物：

。在式Ia化合物之一較佳組態中，R¹⁵ 為甲基。在式Ia化合物之另一較佳組態中，R¹⁵ 為苄基。在式Ia化合物之一代表性組態中， R¹ 為NH₂ ； R² 為H； R¹³ 為苯基、萘基或吲哚基，其中任一者視情況經鹵基(例如溴)或C₃ -C₄ 環烷基(例如環丙基)取代； R¹⁵ 為C₁ -C₃ 烷基； R¹⁶ 為C₁ -C₈ 烷基。在式Ia化合物之另一代表性組態中， R¹ 為NH₂ ； R² 為H； R¹³ 為萘基； R¹⁵ 為C₁ -C₃ 烷基； R¹⁶ 為C₁ -C₈ 烷基或苄基。在式Ia化合物之另一代表性組態中， R¹ 為NH₂ ； R² 為H； R¹³ 為苯基，其4-位視情況經鹵基(例如溴)或經C₃ -C₄ 環烷基(例如環丙基)取代； R¹⁵ 為甲基； R¹⁶ 為C₃ -C₈ 烷基。在式Ia化合物之另一代表性組態中， R¹ 為NH₂ ； R² 為H； R¹³ 為苯基； R¹⁵ 為甲基； R¹⁶ 為C₃ -C₈ 烷基。在式Ia化合物之另一代表性組態中， R¹ 為NH₂ ； R² 為F； R¹³ 為苯基、萘基或吲哚基，其中任一者視情況經鹵基(例如溴)或C₃ -C₄ 環烷基(例如環丙基)取代； R¹⁵ 為C₁ -C₃ 烷基； R¹⁶ 為C₁ -C₈ 烷基。在式Ia化合物之另一代表性組態中， R¹ 為NH₂ ； R² 為F； R¹³ 為萘基； R¹⁵ 為C₁ -C₃ 烷基； R¹⁶ 為C₁ -C₈ 烷基或苄基。在式Ia化合物之另一代表性組態中， R¹ 為NH₂ ； R² 為F； R¹³ 為苯基，其4-位視情況經鹵基(例如溴)或經C₃ -C₄ 環烷基(例如環丙基)取代； R¹⁵ 為甲基； R¹⁶ 為C₃ -C₈ 烷基。在式Ia化合物之另一代表性組態中， R¹ 為NH₂ ； R² 為F； R¹³ 為苯基； R¹⁵ 為甲基； R¹⁶ 為C₃ -C₈ 烷基。在另一組態中，R¹⁵ 及R^15' 連同其所連接的碳原子形成C₃ -C₇ 環烷基，例如環丙基或環丁基。 R¹⁶ 一般為C₁ -C₁₀ 烷基或C₃ -C₇ 環烷基。 R¹⁶ 之代表性含義包括C₁ -C₃ 烷基，諸如甲基、乙基、丙基、異丙基。R¹⁶ 之較佳含義為甲基，R¹⁶ 之另一較佳含義為異丙基。在一個實施例中，R¹⁶ 為C₃ -C₁₀ 烷基。根據此實施例，R¹⁶ 之代表性含義包括分支鏈C₅ -C₈ 烷基。在一個實施例中，R¹⁶ 之分支點在C₁ 處。在一替代性實施例中，R¹⁶ 之分支點在C² 處。典型地，根據此等實施例，R^15' 為H，R^15' 所連接之碳原子之立體化學為L-胺基酸之立體化學，因此提供以下通式之化合物：

其中R¹⁶¹ 及R¹⁶² 為相同或不同的C₁ -C₃ 烷基，且R¹⁶³ 及R¹⁶⁴ 為相同或不同的C₁ -C₃ 烷基。典型地，在式(Ia')化合物中，R¹⁶ 為2-戊基，即R¹⁶¹ 為丙基且R¹⁶² 為甲基。在式(Ia')化合物之另一典型組態中，R¹⁶ 為2-丁基，即R¹⁶¹ 為乙基且R¹⁶² 為甲基。典型地，在式(Ia")化合物中，R¹⁶ 為2-丙基戊基或2-乙基丁基，即R¹⁶³ 及R¹⁶⁴ 皆分別為丙基或乙基。 R¹⁶ 之其他代表性含義包括C₃ -C₇ 環烷基，例如環己基。 R¹⁶ 之另一代表性含義為環戊基。 R¹⁶ 之另一代表性含義為苄基。 R¹³ 一般為苯基、萘基或吲哚基，其中任一者視情況經1或2個R²² 取代。在本發明之一個實施例中，R¹³ 為苯基或萘基，其中任一者視情況經取代。在本發明之一個實施例中，R¹³ 為萘基。在本發明之一較佳實施例中，R¹³ 為苯基。 R¹³ 之代表性實例包括苯基，其視情況經一個、兩個或三個R²² 取代，因此提供式(II-aa)化合物：

其中各R²² 存在時獨立地選自鹵基、C₁ -C₆ 烷基、C₂ -C₆ 烯基及C₁ -C₆ 烷氧基。典型地，苯環未經取代或經一個R²² 取代。在式(II-aa)化合物之一個組態中，苯環未經取代。在式(II-aa)化合物之另一組態中，苯環經一個R²² 取代。典型地，在此組態中，取代基R²² 位於苯環之4-位。在本發明化合物之一個實施例中，R¹³ 為苯基，其4-位經鹵基(例如溴)或經C₃ -C₄ 環烷基(例如環丙基)取代。在式(II-aa)化合物之一個組態中，苯環經羧基C₁ -C₆ 烷基取代。此組態之一代表性實例以部分式說明：

。在式(II-aa)化合物之另一組態中，苯環經位於相鄰碳原子上之兩個R²² 取代且兩個R²² 組合形成-O-CH₂ -O-，因此形成部分結構：

。 R¹³ 之其他代表性含義包括視情況經取代之吡啶基。典型地，吡啶基部分未經取代或經一或兩個各自獨立地選自鹵基、C₁ -C₆ 鹵烷基、C₁ -C₆ 烷基、C₂ -C₆ 烯基、C₁ -C₆ 烷氧基、羥基、胺基的取代基取代。在式(I)化合物之一典型實施例中， R¹ 為NH₂ 或NHC(=O)C₁ -C₆ 烷基； R¹³ 為苯基、萘基或吲哚基，其中任一者視情況經鹵基、C₁ -C₃ 烷基、C₁ -C₃ 烷氧基、C₃ -C₆ 環烷基或C₁ -C₃ 鹵烷基取代； R^15' 為H且R¹⁵ 為C₁ -C₃ 烷基或苄基； R¹⁶ 為C₁ -C₁₀ 烷基或C₃ -C₇ 環烷基。在式(I)或(Ia)化合物之一典型實施例中， R¹ 為NH₂ 或NHC(=O)C₁ -C₆ 烷基； R¹³ 為苯基或萘基，其中任一者視情況經鹵基、C₁ -C₃ 烷基、C₁ -C₃ 烷氧基、C₃ -C₆ 環烷基或C₁ -C₃ 鹵烷基取代； R^15' 為H且R¹⁵ 為C₁ -C₃ 烷基或苄基； R¹⁶ 為C₂ -C₁₀ 烷基或C₃ -C₇ 環烷基。在式(I)化合物之另一典型實施例中， R¹ 為NH₂ ； R² 為H； R¹³ 為苯基； R^15' 為H且R¹⁵ 為C₁ -C₃ 烷基； R¹⁶ 為C₁ -C₃ 烷基或環己基。在式(I)或(Ia)化合物之另一典型實施例中， R¹ 為NH₂ ； R² 為H； R¹³ 為苯基； R^15' 為H且R¹⁵ 為C₁ -C₃ 烷基或苄基； R¹⁶ 為C₃ -C₈ 烷基、環戊基或環己基。本發明化合物顯示針對癌症(尤其肝癌，諸如HCC)之活性，並可用作治療患有癌症之溫血動物(尤其人類)之藥劑。化合物尤其可用作治療患有肝癌(例如HCC)之人類之藥劑。為了避免所非需之副作用，尤其在其他器官中之毒性，將藥物遞送到腫瘤位置同時減少暴露於正常組織係至關重要的。因為本發明化合物在胃液中係穩定的但易被肝酶代謝，所以其可在胃中被吸收並以經遮蔽之細胞毒性劑輸送到肝臟，在肝臟中吸收、代謝及形成活性細胞毒性三磷酸。因此，本發明提供主要在肝臟中吸收及加工之化合物，因此最大程度地減少暴露於體內其他器官及毒副作用。不希望受理論限制，本發明化合物之抗致癌活性可直接作用於癌症之快速作用的致瘤細胞之細胞過程，但是可額外地或替代地藉由調節腫瘤微環境發揮其作用，例如抑制血管生成，從而餓養供給之腫瘤，導致腫瘤生長之抑制。本發明化合物亦適用於治療繼發性肝癌、肝轉移，即源自體內別處之器官(諸如結腸、肺或乳腺)並遷移到肝之癌症。本發明亦關於一種治療患有癌症，尤其肝癌(諸如HCC)之溫血動物，尤其人類之方法，所述方法包括投與有效量之式(I)化合物或其任意子組。本發明亦關於一種治療患有繼發性肝癌之溫血動物，尤其人類之方法，所述方法包括投與有效量之式(I)化合物或其任意子組。用作藥劑或治療方法之該用途包括向癌症個體全身性投與有效量之式(I)化合物。在一個態樣中，本發明提供一種醫藥組合物，其包含式(I)化合物以及醫藥學上可接受之佐劑、稀釋劑、賦形劑或載劑。在另一態樣中，本發明提供一種用於治療癌症之醫藥組合物，其包含式(I)化合物以及醫藥學上可接受之佐劑、稀釋劑、賦形劑或載劑。在另一態樣中，本發明提供一種用於治療肝癌(諸如HCC)之醫藥組合物，其包含式(I)化合物以及醫藥學上可接受之佐劑、稀釋劑、賦形劑或載劑。在另一態樣中，本發明提供一種用於治療繼發性肝癌之醫藥組合物，其包含式(I)化合物以及醫藥學上可接受之佐劑、稀釋劑、賦形劑或載劑。在另一態樣中，本發明係關於一種用於製備文中所述之醫藥組合物之方法，其包括將醫藥學上可接受之佐劑、稀釋劑、賦形劑及/或載劑與治療有效量之式(I)化合物充分混合。在另一態樣中，本發明提供一種用於如上所述之治療或抑制之醫藥組合物，其進一步包括一或多種額外治療劑。雖然上述醫藥組合物一般包含有效量(例如，對人類而言)之式(I)化合物，但是當與其他藥劑組合使用或多劑量使用時式(I)化合物之亞治療量可能有價值。在此上下文中，治療有效量為足以產生預期結果之量。治療有效量將視在各特定情況下之個人需求而變化。影響劑量之特徵為例如所治療疾病之嚴重程度、所治療個體之年齡、體重、總體健康情況等。關於抗癌效果，此效果可為抑制腫瘤進一步生長，減少轉移可能性或消除轉移或導致腫瘤之細胞死亡，導致腫瘤收縮或防止在患者腫瘤處於緩解期後腫瘤再生長。在另一態樣中，本發明提供用作藥劑之式(I)化合物。在另一態樣中，本發明提供用於治療癌症之式(I)化合物。在另一態樣中，本發明提供用於治療肝癌(諸如HCC)之式(I)化合物。在另一態樣中，本發明提供用於治療繼發性肝癌之式(I)化合物。在另一態樣中，本發明提供用於如上所述治療之式(I)化合物，與一或多種額外癌症治療組合，諸如其他抗癌藥、手術、免疫療法及/或局部療法，如射頻消融。在另一實施例中，另一抗癌治療為放射療法。在一個實施例中，另一抗癌治療為一或多種表現出有效的抗腫瘤活性之其他核苷類似物。在一個態樣中，本發明提供一種醫藥組合，其包含治療有效量之式(I)化合物及一或多種選自由化療劑、多抗藥性逆轉劑及生物應答調節劑組成之群之額外治療劑。在此態樣之一個實施例中，額外治療劑為化療劑。在另一態樣中，本發明提供用於製備藥劑之式(I)化合物。在另一態樣中，本發明提供用於製備治療癌症用之藥劑之式(I)化合物。在另一態樣中，本發明提供用於製備治療肝癌(諸如HCC)用之藥劑之式(I)化合物。在另一態樣中，本發明提供用於製備治療繼發性肝癌用之藥劑之式(I)化合物。在另一態樣中，本發明提供一種用於治療癌症之方法，其包括向有需要之個體(例如人類)投與治療有效量之式(I)化合物。在另一態樣中，本發明提供一種用於治療肝癌(諸如HCC)之方法，其包括向有需要之個體(例如人類)投與治療有效量之式(I)化合物。在另一態樣中，本發明提供一種用於治療繼發性肝癌之方法，其包括向有需要之個體(例如人類)投與治療有效量之式(I)化合物。在另一態樣中，本發明提供一種用於如上所述治療之方法，與額外癌症治療組合，諸如其他抗癌藥、手術、免疫療法及/或局部療法，如射頻消融。在一個態樣中，本發明提供一種用於治療原發性或繼發性肝癌之方法，其包括投與醫藥組合，該組合包含治療有效量之式I化合物，進一步包含一或多種選自由化療劑、多抗藥性逆轉劑及生物反應調節劑組成之群之額外治療劑。在此態樣之一個實施例中，其他治療劑為化療劑。在一個態樣中，本發明提供選自如下所示之化合物之式(I)化合物：

或其醫藥學上可接受之鹽。此外，本發明係關於一種用於製備式(I)化合物之方法，用於製備式(I)化合物之新穎中間物以及該等中間物之製備。當術語『式(I)化合物』、『本發明化合物』或類似術語用於上文及下文時，其意指包括式(I)化合物以及式(I)化合物之任意子組，包括所有可能之立體化學異構形式，其醫藥學上可接受之鹽、溶劑合物、四級胺及金屬錯合物。可將本發明化合物調配成各種醫藥形式用於投藥目的。作為適宜組合物，可提及常用於經口投與藥物之所有組合物。為了製備本發明之醫藥組合物，使有效量之作為活性成分之特定化合物(視情況以加成鹽形式或溶劑合物)與醫藥學上可接受之載劑以均勻混合物組合，該載劑可採用多種形式，此取決於投藥所需之製劑形式。此等醫藥組合物宜呈適於經口投藥之單位劑型。舉例而言，在製備經口劑型之組合物中，可使用任意常見之醫藥介質，諸如，在經口液體製劑(諸如懸浮液、糖漿、酏劑、乳液及溶液)之情況下，例如水、二醇、油、醇及其類似物；或在粉末、丸劑、膠囊及錠劑之情況下，固體載劑，諸如澱粉、糖、高嶺土、潤滑劑、黏合劑、崩解劑及其類似物。錠劑及膠囊因為易投與所以代表最有利之經口劑量單位形式，在此情況下，顯然使用固體醫藥載劑。亦包括在臨用前轉化為液體形式製劑之固體形式製劑。尤其宜調配單位劑型之上述醫藥組合物以便於使用及劑量均勻。文中所用之單位劑型係指適宜作為單位劑量之實體離散單位，各單位含有經計算以產生所需療效之預定量之活性成分以及所需醫藥載劑。該等單位劑型之實例為錠劑(包括壓痕或包衣錠劑)、膠囊、丸劑、粉末包、薄片及其類似物，以及其分開之多份。一般而言，預期癌症日有效量為約0.01至約700 mg/kg體重、或約0.5至約400 mg/kg體重、或約1至約250 mg/kg體重、或約2至約200 mg/kg體重、或約10至約150 mg/kg體重。可能宜在一天內以適當的間隔分兩個、三個、四個或多個子劑量來投與所需劑量。所述子劑量可調配成單位劑型，例如，每單位劑型含有約1至約5000 mg、或約50至約3000 mg、或約100至約1000 mg，或約200至約600 mg，或約100至約400 mg活性成分。本發明化合物可僅表現出抗癌作用及/或提高另一抗癌劑表現出抗癌作用之能力。本發明化合物表現為定義之立體異構體。該等化合物之絕對組態可利用技術已知之方法確定，諸如，X-射線繞射或NMR及/或自具有已知立體化學之起始材料推斷。根據本發明之醫藥組合物將較佳包含所述立體異構體之實質上立體異構純之製劑。文中所述之化合物及中間物之純立體異構形式定義為實質上不含該等化合物或中間物之相同的基本分子結構之其他對映異構或非對映異構形式。特定言之，術語「立體異構純的」涉及具有至少80%之立體異構過量(即，最少90%之一種異構體以及最多10%之其他可能的異構體)至100%之立體異構過量(即，100%之一種異構體不含其他)之化合物或中間物，更特定言之，具有90%至100%之立體異構過量之化合物或中間物，甚至更特定言之，具有94%至100%之立體異構過量以及最特定言之，具有97%至100%之立體異構過量。術語「對映異構純的」及「非對映異構純的」應以類似方式理解，但分別關於所討論之混合物之對映異構過量以及非對映異構過量。本發明化合物及中間物之純立體異構形式可藉由應用技術已知程序而得到。舉例而言，可藉由利用光學活性酸或鹼選擇性結晶其等非對映異構鹽而將對映異構體彼此分離。其實例為酒石酸、二苯甲醯基酒石酸、二甲苯醯基酒石酸及樟腦磺酸。或者，對映異構體可藉由層析技術利用對掌性固定相分離。該等純的立體化學異構形式亦可由適宜起始物之相應的純的立體化學異構形式得到，條件為反應以立體特異性方式進行。較佳地，如果需要特定立體異構體，該化合物係藉由立體特異性製備方法合成。此等方法宜使用對映異構純的起始物。本發明化合物之非對映異構外消旋體可單獨地藉由習知方法得到。宜使用之適宜的物理分離方法為例如選擇性結晶及層析，例如管柱層析。當磷原子存在於本發明化合物中時，磷原子可表示對掌性中心。根據Cahn-Ingold-Prelog優先規則，該中心之對掌性表示為「R」或「S」。當沒有指出對掌性時，預期意指包括R-及S-異構體，以及兩者之混合物，即非對映異構混合物。在本發明之較佳實施例中，包括磷原子具有S-組態之立體異構體。此等立體異構體表示為S_P 。在本發明之其他實施例中，包括磷原子具有R-組態之立體異構體。此等立體異構體表示為R_P 。在本發明之其他實施例中，包括非對映異構混合物，即磷原子具有R-或S-組態之化合物之混合物。本發明亦包括同位素標記之式(I)化合物，其中一或多個原子經該原子之同位素，即具有與自然中常見之原子相同的原子序數，但不同原子量之原子，置換。可併入式(I)化合物之同位素之實例包括但不限於氫的同位素，諸如² H及³ H(亦分別用D表示氘以及用T表示氚)；碳，諸如¹¹ C、¹³ C及¹⁴ C；氮，諸如¹³ N及¹⁵ N；氧，諸如¹⁵ O、¹⁷ O及¹⁸ O；磷，諸如³¹ P及³² P；硫，諸如³⁵ S；氟，諸如¹⁸ F；氯，諸如³⁶ Cl；溴，諸如⁷⁵ Br、⁷⁶ Br、⁷⁷ Br及⁸² Br；以及碘，諸如¹²³ I、¹²⁴ I、¹²⁵ I及¹³¹ I。同位素標記之化合物中所包括之同位素之選擇將取決於該化合物之特定應用。舉例而言，對於藥物或基質組織分佈分析，其中併入諸如³ H或¹⁴ C之放射性同位素之化合物一般最有用。對於放射成像應用，例如正電子發射斷層攝影術(PET)，諸如¹¹ C、¹⁸ F、¹³ N或¹⁵ O之正電子發射同位素將為有用的。併入較重同位素(諸如氘，即² H)可向式(I)化合物提供更大的代謝穩定性，此可例如增加化合物之體內半衰期或減少劑量需求。本發明之同位素標記之化合物可藉由與下文中流程及/或實例中所述者類似之方法，利用適宜之同位素標記之試劑或起始物替代相應的非同位素標記之試劑或起始物，或藉由技術人員已知之習知技術製備。醫藥學上可接受之加成鹽包含式(I)化合物之治療活性酸及鹼加成鹽形式。所關注的為式(I)化合物或其子組之游離(即非鹽)形式。醫藥學上可接受之酸加成鹽宜藉由利用該適宜酸處理鹼形式得到。適宜酸包括，例如，無機酸，諸如氫鹵酸，例如鹽酸或氫溴酸、硫酸、硝酸、磷酸及類似酸；或有機酸，諸如乙酸、丙酸、羥基乙酸、乳酸、丙酮酸、草酸(即乙二酸)、丙二酸、琥珀酸(即丁二酸)、順丁烯二酸、反丁烯二酸、蘋果酸(即羥基丁二酸)、酒石酸、檸檬酸、甲磺酸、乙磺酸、苯磺酸、對甲苯磺酸、環己胺磺酸、水楊酸、對胺基水楊酸、雙羥萘酸以及類似酸。相反地，該等鹽形式可藉由適宜鹼處理轉化成游離鹼形式。含有酸性質子之式(I)化合物亦可藉由適宜有機及無機鹼處理而轉化為其等非毒性金屬或胺加成鹽形式。適宜鹼鹽形式包含，例如銨鹽、鹼及鹼土金屬鹽，例如鋰、鈉、鉀、鎂、鈣鹽及其類似物，與有機鹼之鹽，例如苄星、N-甲基-D-葡糖胺、海卓胺，以及與胺基酸之鹽，諸如，精胺酸、賴胺酸及其類似物。一些式(I)化合物亦可以其互變異構形式存在。舉例而言，醯胺基(-C(=O)-NH-)之互變異構形式為亞胺基醇(-C(OH)=N-)，其在具有芳香性之環中變得穩定。所述形式雖然在文中所示之結構式中沒有明確指出，但意欲包括在本發明範疇內。在摘要、說明書及申請專利範圍中所用之術語及表述應按照如下定義理解，除非另外指出。各術語之含義在每次出現時係獨立的。此等定義無論術語單獨使用抑或與其他術語組合使用均適用，除非另外指出。文中使用之未明確定義之術語或表述應理解為具有其在該領域中所用之常見含義。化學名稱、普通名稱及化學結構可互換使用來描述同一結構。如果化合物使用化學結構及化學名稱兩者來指稱且結構與名稱之間存在歧義，則以結構為準。「C_m -C_n 烷基」自身或在諸如C_m -C_n 鹵烷基、C_m -C_n 烷基羰基、C_m -C_n 烷基胺等之複合表述中表示具有指定碳原子數之直鏈或分支鏈脂族烴基，例如C₁ -C₄ 烷基意指具有1至4個碳原子之烷基。C₁ -C₆ 烷基具有相應含義，亦包括戊基及己基之所有直鏈及分支鏈異構體。用於本發明之較佳烷基為C₁ -C₆ 烷基，包括甲基、乙基、正丙基、異丙基、正丁基、異丁基、第二丁基、第三丁基、正戊基及正己基，尤其C₁ -C₄ 烷基，諸如甲基、乙基、正丙基、異丙基、第三丁基、正丁基及異丁基。甲基及異丙基一般為較佳的。烷基可未經取代或經一或多個可能相同或不同的取代基取代，各取代基獨立地選自由以下組成之群：鹵基、烯基、炔基、芳基、環烷基、氰基、羥基、-O-烷基、-O-芳基、-伸烷基-O-烷基、烷硫基、-NH₂ 、-NH(烷基)、-N(烷基)₂ 、-NH(環烷基)、-O-C(=O)-烷基、-O-C(=O)-芳基、-O-C(=O)-環烷基、-C(=O)OH以及-C(=O)O-烷基。一般較佳的為烷基未經取代，除非另外指出。「C₂ -C_n 烯基」表示含有至少一個碳碳雙鍵並具有指定碳原子數之直鏈或分支鏈脂族烴基，例如，C₂ -C₄ 烯基意指具有2至4個碳原子之烯基；C₂ -C₆ 烯基意指具有2至6個碳原子之烯基。非限制性烯基包括乙烯基、丙烯基、正丁烯基、3-甲基丁-2-烯基、正戊烯基以及己烯基。烯基可未經取代或經一或多個可能相同或不同的取代基取代，各取代基獨立地選自由以下組成之群：鹵基、烯基、炔基、芳基、環烷基、氰基、羥基、-O-烷基、-O-芳基、-伸烷基-O-烷基、烷硫基、-NH₂ 、-NH(烷基)、-N(烷基)₂ 、-NH(環烷基)、-O-C(=O)-烷基、-O-C(=O)-芳基、-O-C(=O)-環烷基、-C(=O)OH以及-C(=O)O-烷基。一般較佳的為烯基未經取代，除非另外指出。「C₂ -C_n 炔基」表示含有至少一個碳碳三鍵並具有指定碳原子數之直鏈或分支鏈脂族烴基，例如C₂ -C₄ 炔基意指具有2至4個碳原子之炔基；C₂ -C₆ 炔基意指具有2至6個碳原子之炔基。非限制性炔基包括乙炔基、丙炔基、2-丁炔基及3-甲基丁炔基、戊炔基以及己炔基。炔基可未經取代或經一或多個可能相同或不同的取代基取代，各取代基獨立地選自由以下組成之群：鹵基、烯基、炔基、芳基、環烷基、氰基、羥基、-O-烷基、-O-芳基、-伸烷基-O-烷基、烷硫基、-NH₂ 、-NH(烷基)、-N(烷基)₂ 、-NH(環烷基)、-O-C(=O)-烷基、-O-C(=O)-芳基、-O-C(=O)-環烷基、-C(=O)OH以及-C(=O)O-烷基。一般較佳的為炔基未經取代，除非另外指出。如文中所用，術語「C_m -C_n 鹵烷基」表示其中至少一個C原子經鹵素(例如，C_m -C_n 鹵烷基可含有一至三個鹵素原子)，較佳氯或氟取代之C_m -C_n 烷基。典型的鹵烷基為C₁ -C₂ 鹵烷基，其中鹵基宜表示氟。示例性鹵烷基包括氟甲基、二氟甲基及三氟甲基。如文中所用，術語「C_m -C_n 羥烷基」表示其中至少一個C原子經一個羥基取代之C_m -C_n 烷基。典型的C_m -C_n 羥烷基為其中一個C原子經一個羥基取代之C_m -C_n 烷基。示例性羥烷基包括羥甲基及羥乙基。如文中所用，術語「C_m -C_n 胺基烷基」表示其中至少一個C原子經一個胺基取代之C_m -C_n 烷基。典型的C_m -C_n 胺基烷基為其中一個C原子經一個胺基取代之C_m -C_n 烷基。示例性胺基烷基包括胺甲基及胺乙基。如文中所用，術語「C_m -C_n 伸烷基」表示具有指定碳原子數之直鏈或分支鏈二價烷基。用於本發明之較佳C_m -C_n 伸烷基為C₁ -C₃ 伸烷基。伸烷基之非限制實例包括-CH₂ -、-CH₂ CH₂ -、-CH₂ CH₂ CH₂ -、-CH(CH₃ )CH₂ CH₂ -、-CH(CH₃ )-及-CH(CH(CH₃ )₂ )-。術語「Me」意指甲基，「MeO」意指甲氧基。術語「C_m -C_n 烷基羰基」表示其中C_m -C_n 烷基部分如上定義之式C_m -C_n 烷基-C(=O)-之基團。典型地，「C_m -C_n 烷基羰基」為C₁ -C₆ 烷基-C(=O)-。「C_m -C_n 烷氧基」表示其中C_m -C_n 烷氧如上定義之基團C_m -C_n 烷基-O-。特別關注的是C₁ -C₄ 烷氧基，其包括甲氧基、乙氧基、正丙氧基、異丙氧基、第三丁氧基、正丁氧基及異丁氧基。甲氧基及異丙氧基一般為較佳的。C₁ -C₆ 烷氧基具有相應含義，擴展到包括戊氧基及己氧基之所有直鏈及分支鏈異構體。術語「C_m -C_n 烷氧基羰基」表示其中C_m -C_n 烷氧基如上定義之式C_m -C_n 烷氧基-C(=O)-之基團。典型地，「C_m -C_n 烷氧基羰基」為C₁ -C₆ 烷氧基-C(=O)-。術語「胺基」表示基團-NH₂ 。術語「鹵」表示鹵素基團，例如氟、氯、溴或碘。典型地，鹵基為氟或氯。術語「芳基」意指苯基、聯苯基或萘基。術語「雜環烷基」表示含有1-3個獨立地選自O、S及N之雜原子之穩定飽和單環3-7員環。在一個實施例中，穩定的飽和單環3-7員環含有1個選自O、S及N之雜原子。在第二實施例中，穩定的飽和單環3-7員環含有2個獨立地選自O、S及N之雜原子。在第三實施例中，穩定的飽和單環3-7員環含有3個獨立地選自O、S及N之雜原子。含有1-3個獨立地選自O、S及N之雜原子之穩定的飽和單環3-7員環一般為5-7員環，例如5或6員環。雜環烷基可未經取代或經一或多個可能相同或不同的取代基取代，各取代基獨立地選自由以下組成之群：鹵基、烯基、炔基、芳基、環烷基、氰基、羥基、-O-烷基、-O-芳基、-伸烷基-O-烷基、烷硫基、-NH₂ 、-NH(烷基)、-N(烷基)₂ 、-NH(環烷基)、-O-C(=O)-烷基、-O-C(=O)-芳基、-O-C(=O)-環烷基、-C(=O)OH以及-C(=O)O-烷基。一般較佳的為雜環烷基未經取代，除非另外指出。術語「雜芳基」表示含有1-4個獨立地選自O、S及N之雜原子之穩定的單環或雙環芳族環系統，各環具有5或6個環原子。在本發明之一個實施例中，穩定的單環或雙環芳族環系統含有一個選自O、S及N之雜原子，各環具有5或6個環原子。在本發明之第二實施例中，穩定的單環或雙環芳族環系統含有兩個獨立地選自O、S及N之雜原子，各環具有5或6個環原子。在第三實施例中，穩定的單環或雙環芳族環系統含有三個獨立地選自O、S及N之雜原子，各環具有5或6個環原子。在第四實施例中，穩定的單環或雙環芳族環系統含有四個獨立地選自O、S及N之雜原子，各環具有5或6個環原子。雜芳基之一個實施例包括黃酮。術語「C₃ -C_n 環烷基」表示具有指定碳原子數之環狀單價烷基，例如C₃ -C₇ 環烷基意指具有3至7個碳原子之環狀單價烷基。用於本發明之較佳環烷基為C₃ -C₄ 烷基，即環丙基及環丁基。環烷基可未經取代或經一或多個可能相同或不同的取代基取代，各取代基獨立地選自由以下組成之群：鹵基、烯基、炔基、芳基、環烷基、氰基、羥基、-O-烷基、-O-芳基、-伸烷基-O-烷基、烷硫基、-NH₂ 、-NH(烷基)、-N(烷基)₂ 、-NH(環烷基)、-O-C(=O)-烷基、-O-C(=O)-芳基、-O-C(=O)-環烷基、-C(=O)OH以及-C(=O)O-烷基。一般較佳的為環烷基未經取代，除非另外指出。術語「胺基C_m -C_n 烷基」表示經胺基取代之如上定義之C_m -C_n 烷基，即烷基中之一個氫原子經NH₂ -基團置換。典型地，「胺基C_m -C_n 烷基」為胺基C₁ -C₆ 烷基。術語「胺基C_m -C_n 烷基羰基」表示如上定義之C_m -C_n 烷基羰基，其中烷基中之一個氫原子經NH₂ -基團置換。典型地，「胺基C_m -C_n 烷基羰基」為胺基C₁ -C₆ 烷基羰基。胺基C_m -C_n 烷基羰基之實例包括但不限於甘胺醯基：C(=O)CH₂ NH₂ 、丙胺醯基：C(=O)CH(NH₂ )CH₃ 、纈胺醯基：C=OCH(NH₂ )CH(CH₃ )₂ 、白胺醯基：C(=O)CH(NH₂ )(CH₂ )₃ CH₃ 、異白胺醯基：C(=O)CH(NH₂ )CH(CH₃ )(CH₂ CH₃ )及正白胺酸基：C(=O)CH(NH₂ )(CH₂ )₃ CH₃ 及其類似基團。此定義不限於天然存在的胺基酸。如文中所用，術語「(=O)」當連接到碳原子時形成羰基部分。應注意，當原子價允許時，一個原子只能攜載一個側氧基。術語「單磷酸酯、二磷酸酯及三磷酸酯」係指如下基團：

及

。如文中所用，定義中所用之任意分子部分上之基團位置可為該部分上之任意處，主要其化學上穩定。當存在之任何變數在任何部分中多次出現時，各定義係獨立的。術語「溶劑合物」涵蓋式(I)化合物以及其鹽能夠形成之任何醫藥學上可接受之溶劑合物。所述溶劑合物為例如水合物、醇化物，例如乙醇化物、丙醇化物及其類似物，尤其水合物。如文中所用，術語「前藥」表示向個體投與後在體內易於藉由代謝及/或化學過程轉化以得到活性化合物之藥物前驅體。如文中所用，表述「肝靶向前藥」表示主要在肝內代謝成其活性物質之前藥。如文中所用，表述「肝癌」意欲包括原發性及繼發性肝癌，即分別地為源自肝臟之癌症，以及來自其他器官之癌症之肝轉移。相關術語應結合以上提供之定義及技術領域之常見用途理解。一般而言，用於本申請案中之化合物之名稱係利用ChemDraw Ultra 12.0.產生。此外，如果結構或結構之一部分之立體化學未經例如加粗或虛線表示，則該結構或該結構之部分應理解成涵蓋其所有立體異構體。一般合成方法 本發明可藉由多種方法製備，例如如以下所示以及所述之說明性合成流程中所述者。所有起始物及試劑可購自供應商或可根據參考文獻中所述之文獻程序利用熟習此項技術者熟知之方法製備。流程1說明式(I)化合物之一般路徑。

流程 1 如上所述製備之市售曲沙他濱衍生物(1a)與所需胺基磷酸酯試劑(1b)(其中Lg為適宜的離去基團，例如鹵素(如氯)或活性苯酚(如五氯苯酚、對硝基苯酚、五氟苯酚或其類似物))在惰性溶劑(諸如醚，例如乙醚或THF)，或鹵化烴(例如二氯甲烷)中，在鹼(諸如N-甲基咪唑(NMI))或格林納試劑(Grignard reagent)(如氯化第三丁基鎂或其類似物)存在下縮合得到胺基磷酸酯衍生物(1c)。用於以上流程之胺基磷酸酯試劑(1b)(其中Lg為氯，即氯化胺基磷酸酯)可以三氯氧磷(POCl₃ )為起始物按照流程2所述以兩步反應製備。

流程 2 POCl₃ 與所需醇R¹³ OH在惰性溶劑(如Et₂ O)中縮合得到烷氧基或芳氧基二氯化磷酸酯(2a)。隨後與胺基酸衍生物(2b)反應得到其中R^3' 為H之氯胺基磷酸酯(2c)。如需要，可如流程3所概述，將獲得之氯胺基磷酸酯(2c)轉化為具有活性苯酚作為離去基團(例如五氟苯酚或對-NO₂ -苯酚)之相應的磷酸化劑。

流程 3 此轉化宜藉由氯衍生物(2c)與所需之活性苯酚在鹼(如三乙胺或類似物)存在下反應由此提供磷酸化劑(3a)及(3b)進行。用於以上流程之多種保護基(PG)之使用對熟習此項技術者而言為已知的，且其應用及另外替代物在文獻中詳述，參見例如Greene T.W., Wuts P.G.M. Protective groups in organic synthesis, 第2版 New York: Wiley; 1995。如文中所用，術語「N-保護基」或「N-經保護的」係指意欲保護胺基酸或肽之N-端或保護胺基酸免於在合成程序期間發生非所需反應之彼等基團。常用之N-保護基團如Greene中所揭示。N-保護基團包括醯基，諸如甲醯基、乙醯基、丙醯基、特戊醯基、第三丁基乙醯基、2-氯乙醯基、2-溴乙醯基、三氟乙醯基、三氯乙醯基、鄰苯二甲醯基、鄰硝基苯氧基乙醯基、α-氯丁醯基、苯甲醯基、4-氯苯甲醯基、4-溴苯甲醯基、4-硝基苯甲醯基及其類似基團；磺醯基，諸如苯磺醯基、對甲苯磺醯基及其類似基團；胺基甲酸酯形成基團，諸如苄氧基羰基、對氯苄氧基-羰基、對甲氧基苄氧基羰基、對硝基苄氧基羰基、2-硝基苄氧基羰基、對溴苄氧基羰基、3,4-二甲氧基苄氧基羰基、4-甲氧基苄氧基羰基、2-硝基-4,5-二甲氧基苄氧基羰基、3,4,5-三甲氧基苄氧基羰基、1-(對聯苯基)-1-甲基乙氧基羰基、α,α-二甲基-3,5-二甲氧基苄氧基羰基、二苯甲氧基羰基、第三丁氧基羰基、二異丙基甲氧基羰基、異丙氧基羰基、乙氧基羰基、甲氧基羰基、烯丙氧基羰基、2,2,2-三氯乙氧基羰基、苯氧基羰基、4-硝基苯氧基羰基、芴基-9-甲氧基羰基、環戊氧基羰基、金剛烷氧基羰基、環己氧基羰基、苯硫基羰基及其類似基團；烷基，諸如苄基、三苯基甲基、苄氧基甲基及其類似基團；以及矽烷基，諸如三甲基矽烷基及其類似基團。偏好的N-保護基包括甲醯基、乙醯基、苯甲醯基、特戊醯基、第三丁基乙醯基、苯基磺醯基、苄基(Bz)、第三丁氧基羰基(BOC)以及苄氧基羰基(Cbz)。羥基及/或羧基保護基團亦在Greene上文中詳述並包括醚，諸如甲基醚；經取代之甲基醚，諸如甲氧基甲基、甲硫基甲基、苄氧基甲基、第三丁氧基甲基、2-甲氧基乙氧基甲基及其類似基團；矽烷基醚，諸如三甲基矽烷基(TMS)、第三丁基二甲基矽烷基(TBDMS)、三苄基矽烷基、三苯基矽烷基、第三丁基二苯基矽烷基、三異丙矽烷基及其類似基團；經取代之乙基醚，諸如1-乙氧基甲基、1-甲基-1-甲氧基乙基、第三丁基、烯丙基、苄基、對甲氧基苄基、二苯基甲基、三苯基甲基及其類似基團；芳烷基，例如三苯甲基以及甲哌啶基(pixyl)(9-羥基-9-苯基二苯并哌喃衍生物，尤其氯化物)。酯羥基保護基團包括酯，諸如甲酸酯、苄基甲酸酯、氯乙酸酯、甲氧基乙酸酯、苯氧基乙酸酯、特戊酸酯、金剛酸酯、菜酸酯(mesitoate)、苯甲酸酯及其類似物。碳酸酯羥基保護基團包括甲基乙烯基、烯丙基、桂皮基、苄基及其類似基團。In one aspect, the invention provides a compound represented by formula (I):

R ¹ is OR ¹¹ , or NR ⁵ R ^5' ; R ² is H or F; R ⁵ is H, C ₁ -C ₆ alkyl, OH, C(=O)R ⁶ , OC(=O)R ⁶ Or OC(=O)OR ⁶ ; R ^5′ is H or C ₁ -C ₆ alkyl; R ⁶ is C ₁ -C ₂₂ alkyl or C ₃ -C ₇ cycloalkyl; R ¹¹ is H or C ₁ -C ₆ alkyl; R ¹³ is H, phenyl, pyridyl, benzyl, indolyl or naphthyl, wherein the phenyl, pyridyl, benzyl, indolyl and naphthyl are optionally 1, 2 Or 3 R ²² substitutions; R ¹⁵ is H, C ₁ -C ₆ alkyl, C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl C ₁ -C ₃ alkyl, phenyl, benzyl Or indolyl; R ^15′ is H or C ₁ -C ₆ alkyl; or R ¹⁵ and R ^15′ together with the carbon atom to which they are attached form C ₃ -C ₇ cycloalkyl, wherein each C _1- The C ₆ alkyl group is optionally substituted with a group selected from halogen, OR ¹⁸ and SR ¹⁸ , and each C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl, phenyl and benzyl are optionally Substitution by one or two groups independently selected from C ₁ -C ₃ alkyl, halo and OR ¹⁸ ; R ¹⁶ is H, C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl C ₁ -C ₃ alkyl, benzyl or phenyl, any one of which is independently selected from halo, 1, 2 or 3, as appropriate OR ¹⁸ and N(R ¹⁸ ) ₂ group substitution; each R ^{18 is} independently H, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, or C ₃ -C ₇ cycloalkyl; each R ^{22 is} independently selected from halo, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy Group, phenyl, hydroxyl C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ alkylcarbonyl, C ₃ -C ₆ cycloalkylcarbonyl, carboxyl C ₁ -C ₆ alkyl , Hydroxyl, amine CN and NO ₂ , or any two R ²² groups attached to adjacent ring carbon atoms can be combined to form -O-(CR ²³ R ^23' ) _1-6 -O-; R ²³ and R ^{23' is} independently H or C ₁ -C ₃ alkyl; or a pharmaceutically acceptable salt and/or solvate thereof. In one embodiment, the present invention provides compounds represented by formula I:

Where: R ¹ is OR ¹¹ or NR ⁵ R ^5' ; R ² is H or F; R ⁵ is H, C ₁ -C ₆ alkyl, OH, C(=O)R ⁶ , OC(=O)R ⁶ or OC(=O)OR ⁶ ; R ^5′ is H or C ₁ -C ₆ alkyl; R ⁶ is C ₁ -C ₂₂ alkyl or C ₃ -C ₇ cycloalkyl; R ¹¹ is H or C ₁ -C ₆ alkyl; R ¹³ is H, phenyl, pyridyl, benzyl, indolyl or naphthyl, wherein the phenyl, pyridyl, benzyl, indolyl and naphthyl are optionally 1, 2 or 3 R ²² substitutions; R ¹⁵ is H, C ₁ -C ₆ alkyl, C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl C ₁ -C ₃ alkyl, phenyl, benzyl Group or indolyl group; R ^15′ is H or C ₁ -C ₆ alkyl; or R ¹⁵ and R ^15′ together with the carbon atom to which they are attached form C ₃ -C ₇ cycloalkyl, wherein each C ₁ -C ₆ alkyl is optionally substituted with a group selected from halo, OR ¹⁸ and SR ¹⁸ , and each C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl, phenyl and benzyl The case is substituted with one or two groups independently selected from C ₁ -C ₃ alkyl, halo and OR ¹⁸ ; R ¹⁶ is H, C ₁ -C ₁₀ alkyl, C ₂ -C ₁₀ alkenyl, C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl C ₁ -C ₃ alkyl, benzyl or phenyl, any of which is independently selected from halogen, 1, 2 or 3, as appropriate OR ¹⁸ and N(R ¹⁸ ) ₂ group substitution; each R ^{18 is} independently H, C ₁ -C ₆ alkyl, C ₁ -C ₆ haloalkyl, or C ₃ -C ₇ cycloalkyl; each R ^{22 is} independently selected from halo, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkoxy, C ₁ -C ₆ haloalkoxy Group, phenyl, hydroxyl C ₁ -C ₆ alkyl, C ₃ -C ₆ cycloalkyl, C ₁ -C ₆ alkylcarbonyl, C ₃ -C ₆ cycloalkylcarbonyl, carboxyl C ₁ -C ₆ alkyl , Hydroxyl, amine CN, NO ₂ and trimethylsilyl, or any two R ²² groups attached to adjacent ring carbon atoms can be combined to form -O-(CR ²³ R ^23' ) _1-6 -O -; R ²³ and R ^{23' are} independently H or C ₁ -C ₃ alkyl; or a pharmaceutically acceptable salt and/or solvate thereof. The compound of formula (I) is optionally provided in the form of pharmaceutically acceptable salts and/or solvates. In one embodiment, the compound of the present invention is provided as a pharmaceutically acceptable salt. In the second embodiment, the compound of the present invention is provided as a pharmaceutically acceptable solvate. In a third embodiment, the compound of the present invention is provided in its free form. In a representative embodiment of the invention, R ¹ is NR ⁵ R ^5′ , such as NH ₂ or NHC(=0)C ₁ -C ₆ alkyl. R ^{2 is} generally H. In a preferred embodiment, R ¹ is NH ₂ and R ² is H. In an alternative embodiment, R ¹ is NH ₂ and R ² is F. Typically, in compounds of formula (I), a portion of -NHC(R ¹⁵ )(R ^15' )-C(=O)OR ¹⁶ forms amino acid ester residues, including natural and unnatural amino acid residues. Particular attention is paid to amino acid residues where R ^15' is hydrogen and R ¹⁵ is methyl, isopropyl, isobutyl or benzyl. In a typical configuration, R ^15' is H and R ¹⁵ is C ₁ -C ₃ alkyl, such as methyl, ethyl, propyl, isopropyl. In compounds where R ^15' is hydrogen and R ^{15 is} not hydrogen, the configuration of asymmetric carbon atoms is generally the configuration of L-amino acids, thus providing compounds with the stereochemistry shown in formula (Ia) :

. In one preferred configuration of the compound of formula Ia, R ¹⁵ is methyl. In another preferred configuration of the compound of formula Ia, R ¹⁵ is benzyl. In one representative configuration of the compound of formula Ia, R ¹ is NH ₂ ; R ² is H; R ¹³ is phenyl, naphthyl or indolyl, any of which is optionally halogenated (eg bromine) or C ₃ -C ₄ cycloalkyl (eg cyclopropyl) substitution; R ¹⁵ is C ₁ -C ₃ alkyl; R ¹⁶ is C ₁ -C ₈ alkyl. In another representative configuration of the compound of formula Ia, R ¹ is NH ₂ ; R ² is H; R ¹³ is naphthyl; R ¹⁵ is C ₁ -C ₃ alkyl; R ¹⁶ is C ₁ -C ₈ alkyl Radical or benzyl. In another representative configuration of the compound of formula Ia, R ¹ is NH ₂ ; R ² is H; R ¹³ is phenyl, its 4-position is optionally halogenated (eg bromine) or C ₃ -C ₄ Cycloalkyl (eg cyclopropyl) substitution; R ¹⁵ is methyl; R ¹⁶ is C ₃ -C ₈ alkyl. In another representative configuration of the compound of Formula Ia, R ¹ is NH ₂ ; R ² is H; R ¹³ is phenyl; R ¹⁵ is methyl; R ¹⁶ is C ₃ -C ₈ alkyl. In another representative configuration of the compound of formula Ia, R ¹ is NH ₂ ; R ² is F; R ¹³ is phenyl, naphthyl or indolyl, any of which is optionally halogenated (eg bromine) Or C ₃ -C ₄ cycloalkyl (eg cyclopropyl) substitution; R ¹⁵ is C ₁ -C ₃ alkyl; R ¹⁶ is C ₁ -C ₈ alkyl. In another representative configuration of the compound of formula Ia, R ¹ is NH ₂ ; R ² is F; R ¹³ is naphthyl; R ¹⁵ is C ₁ -C ₃ alkyl; R ¹⁶ is C ₁ -C ₈ alkyl Radical or benzyl. In another representative configuration of the compound of formula Ia, R ¹ is NH ₂ ; R ² is F; R ¹³ is phenyl, the 4-position of which is optionally halogenated (eg bromine) or C ₃ -C ₄ Cycloalkyl (eg cyclopropyl) substitution; R ¹⁵ is methyl; R ¹⁶ is C ₃ -C ₈ alkyl. In another representative configuration of the compound of formula Ia, R ¹ is NH ₂ ; R ² is F; R ¹³ is phenyl; R ¹⁵ is methyl; R ¹⁶ is C ₃ -C ₈ alkyl. In another configuration, R ¹⁵ and R ^15′ together with the carbon atom to which they are attached form a C ₃ -C ₇ cycloalkyl group, such as cyclopropyl or cyclobutyl. R ^{16 is} generally C ₁ -C ₁₀ alkyl or C ₃ -C ₇ cycloalkyl. Representative meanings of R ¹⁶ include C ₁ -C ₃ alkyl groups such as methyl, ethyl, propyl, isopropyl. Preferred meaning of R ¹⁶ is methyl, another preferred meaning of R ¹⁶ is isopropyl. In one embodiment, R ¹⁶ is C ₃ -C ₁₀ alkyl. According to this embodiment, the representative meaning of R ¹⁶ includes branched C ₅ -C ₈ alkyl. In one embodiment, the branch point of R ¹⁶ is at C ₁ . In an alternative embodiment, the branch point of R ¹⁶ is at C ² . Typically, according to these embodiments, R ^15′ is H, and the stereochemistry of the carbon atom to which R ^15′ is attached is the stereochemistry of L-amino acid, thus providing compounds of the general formula:

Wherein R ¹⁶¹ and R ¹⁶² are the same or different C ₁ -C ₃ alkyl groups, and R ¹⁶³ and R ¹⁶⁴ are the same or different C ₁ -C ₃ alkyl groups. Typically, in the compound of formula (Ia'), R ¹⁶ is 2-pentyl, ie R ¹⁶¹ is propyl and R ¹⁶² is methyl. In another typical configuration of the compound of formula (Ia'), R ¹⁶ is 2-butyl, that is, R ¹⁶¹ is ethyl and R ¹⁶² is methyl. Typically, in the compound of formula (Ia"), R ¹⁶ is 2-propylpentyl or 2-ethylbutyl, ie, R ¹⁶³ and R ¹⁶⁴ are respectively propyl or ethyl. Other representatives of R ¹⁶ is meant to include C ₃ -C ₇ cycloalkyl, such as cyclohexyl. another representative of the meaning of R ¹⁶ is cyclopentyl. another representative of the meaning of R ¹⁶ is benzyl. R ¹³ typically is phenyl, naphthyl or Indolyl, either of which is optionally substituted with 1 or 2 R ^22. In one embodiment of the present invention, R ¹³ is phenyl or naphthyl, and any of them is optionally substituted. In one embodiment, R ¹³ is a naphthyl group. In a preferred embodiment of the present invention, R ¹³ is a phenyl group. Representative examples of R ¹³ include a phenyl group, which is optionally substituted by one, two, or three R ²² substitution, thus providing compounds of formula (II-aa):

Where each R ^{22 is} present, it is independently selected from halo, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl and C ₁ -C ₆ alkoxy. Typically, the benzene ring is unsubstituted or substituted with one R ²² . In one configuration of the compound of formula (II-aa), the benzene ring is unsubstituted. In another configuration of the compound of formula (II-aa), the benzene ring is substituted with one R ²² . Typically, in this configuration, the substituent R ²² is located at the 4-position of the benzene ring. In an embodiment of the compound of the present invention, R ¹³ is phenyl, and its 4-position is substituted with a halogen group (eg, bromine) or a C ₃ -C ₄ cycloalkyl group (eg, cyclopropyl). In one configuration of the compound of formula (II-aa), the benzene ring is substituted with carboxyl C ₁ -C ₆ alkyl. A representative example of this configuration is explained in part:

. In another configuration of the compound of formula (II-aa), the benzene ring is substituted by two R ²² located on adjacent carbon atoms and the two R ^{22 are} combined to form -O-CH ₂ -O-, thus forming a partial structure :

. Other representative meanings of R ¹³ include optionally substituted pyridyl. Typically, the pyridyl moiety is unsubstituted or is independently selected from one or two of halo, C ₁ -C ₆ haloalkyl, C ₁ -C ₆ alkyl, C ₂ -C ₆ alkenyl, C ₁ -C ₆ alkoxy, hydroxy, amine substituent substitution. In a typical embodiment of the compound of formula (I), R ¹ is NH ₂ or NHC(=O)C ₁ -C ₆ alkyl; R ¹³ is phenyl, naphthyl or indolyl, any of which is regarded as The case is substituted with halo, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, C ₃ -C ₆ cycloalkyl or C ₁ -C ₃ haloalkyl; R ^15′ is H and R ¹⁵ is C ₁ -C ₃ alkyl or benzyl; R ¹⁶ is C ₁ -C ₁₀ alkyl or C ₃ -C ₇ cycloalkyl. In a typical embodiment of a compound of formula (I) or (Ia), R ¹ is NH ₂ or NHC(=O)C ₁ -C ₆ alkyl; R ¹³ is phenyl or naphthyl, either of which is The case is substituted with halo, C ₁ -C ₃ alkyl, C ₁ -C ₃ alkoxy, C ₃ -C ₆ cycloalkyl or C ₁ -C ₃ haloalkyl; R ^15′ is H and R ¹⁵ is C ₁ -C ₃ alkyl or benzyl; R ¹⁶ is C ₂ -C ₁₀ alkyl or C ₃ -C ₇ cycloalkyl. In another exemplary embodiment of the compound of formula (I), R ¹ is NH ₂ ; R ² is H; R ¹³ is phenyl; R ^15′ is H and R ¹⁵ is C ₁ -C ₃ alkyl; R ¹⁶ It is C ₁ -C ₃ alkyl or cyclohexyl. In another exemplary embodiment of the compound of formula (I) or (Ia), R ¹ is NH ₂ ; R ² is H; R ¹³ is phenyl; R ^15′ is H and R ¹⁵ is C ₁ -C ₃ alkane Radical or benzyl; R ¹⁶ is C ₃ -C ₈ alkyl, cyclopentyl or cyclohexyl. The compounds of the present invention show activity against cancer (especially liver cancer, such as HCC) and can be used as an agent for treating warm-blooded animals (especially humans) with cancer. The compounds are particularly useful as medicaments for treating humans with liver cancer (eg HCC). To avoid undesirable side effects, especially toxicity in other organs, it is essential to deliver drugs to the tumor site while reducing exposure to normal tissue systems. Because the compound of the present invention is stable in gastric juice but easily metabolized by liver enzymes, it can be absorbed in the stomach and transported to the liver as a masked cytotoxic agent, where it is absorbed, metabolized, and forms active cytotoxic triphosphate . Therefore, the present invention provides compounds that are mainly absorbed and processed in the liver, thereby minimizing exposure to other organs in the body and toxic and side effects. Without wishing to be bound by theory, the anti-carcinogenic activity of the compounds of the present invention can directly act on the cellular processes of fast-acting tumorigenic cells of cancer, but can additionally or alternatively exert its effects by regulating the tumor microenvironment, such as inhibiting angiogenesis In order to feed the supplied tumors, the tumor growth is inhibited. The compounds of the present invention are also suitable for the treatment of secondary liver cancer, liver metastasis, ie cancers that originate from organs elsewhere in the body (such as colon, lung or breast) and migrate to the liver. The invention also relates to a method of treating warm-blooded animals, especially humans, suffering from cancer, especially liver cancer (such as HCC), which method comprises administering an effective amount of a compound of formula (I) or any subgroup thereof. The invention also relates to a method for treating warm-blooded animals, especially humans, with secondary liver cancer, which method comprises administering an effective amount of a compound of formula (I) or any subgroup thereof. The use as a medicament or method of treatment includes systemically administering an effective amount of a compound of formula (I) to a cancer individual. In one aspect, the invention provides a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable adjuvant, diluent, excipient or carrier. In another aspect, the present invention provides a pharmaceutical composition for treating cancer, which comprises a compound of formula (I) and a pharmaceutically acceptable adjuvant, diluent, excipient or carrier. In another aspect, the present invention provides a pharmaceutical composition for treating liver cancer (such as HCC), which comprises a compound of formula (I) and a pharmaceutically acceptable adjuvant, diluent, excipient or carrier . In another aspect, the invention provides a pharmaceutical composition for the treatment of secondary liver cancer, which comprises a compound of formula (I) and a pharmaceutically acceptable adjuvant, diluent, excipient or carrier. In another aspect, the present invention relates to a method for preparing a pharmaceutical composition described herein, which comprises combining a pharmaceutically acceptable adjuvant, diluent, excipient and/or carrier with a treatment An effective amount of the compound of formula (I) is thoroughly mixed. In another aspect, the present invention provides a pharmaceutical composition for treatment or inhibition as described above, which further includes one or more additional therapeutic agents. Although the above pharmaceutical composition generally contains an effective amount (eg, for humans) of the compound of formula (I), the sub-therapeutic amount of the compound of formula (I) may be valuable when used in combination with other agents or in multiple doses. In this context, a therapeutically effective amount is an amount sufficient to produce the desired result. The therapeutically effective amount will vary depending on individual needs in each specific situation. The characteristics that affect the dose are, for example, the severity of the disease being treated, the age, weight, and overall health of the individual being treated. Regarding the anti-cancer effect, this effect may be to inhibit the further growth of the tumor, reduce the possibility of metastasis or eliminate the metastasis or cause cell death of the tumor, cause the tumor to contract or prevent the tumor from regrowth after the patient's tumor is in remission. In another aspect, the present invention provides a compound of formula (I) for use as a medicament. In another aspect, the present invention provides compounds of formula (I) for use in the treatment of cancer. In another aspect, the invention provides compounds of formula (I) for use in the treatment of liver cancer, such as HCC. In another aspect, the present invention provides compounds of formula (I) for use in the treatment of secondary liver cancer. In another aspect, the present invention provides compounds of formula (I) for treatment as described above in combination with one or more additional cancer treatments, such as other anti-cancer drugs, surgery, immunotherapy, and/or local therapy, such as radiofrequency Ablation. In another embodiment, another anti-cancer treatment is radiation therapy. In one embodiment, another anti-cancer treatment is one or more other nucleoside analogs that exhibit effective anti-tumor activity. In one aspect, the invention provides a pharmaceutical combination comprising a therapeutically effective amount of a compound of formula (I) and one or more additional therapeutic agents selected from the group consisting of chemotherapeutic agents, multi-drug resistance reversal agents, and biological response modifiers . In one embodiment of this aspect, the additional therapeutic agent is a chemotherapeutic agent. In another aspect, the present invention provides compounds of formula (I) for use in the preparation of medicaments. In another aspect, the present invention provides compounds of formula (I) for use in the preparation of medicaments for the treatment of cancer. In another aspect, the present invention provides a compound of formula (I) for use in the preparation of a medicament for the treatment of liver cancer, such as HCC. In another aspect, the present invention provides compounds of formula (I) for use in the preparation of a medicament for the treatment of secondary liver cancer. In another aspect, the present invention provides a method for treating cancer, which comprises administering a therapeutically effective amount of a compound of formula (I) to an individual in need, such as a human. In another aspect, the present invention provides a method for treating liver cancer, such as HCC, which comprises administering a therapeutically effective amount of a compound of formula (I) to an individual in need, such as a human. In another aspect, the present invention provides a method for treating secondary liver cancer, which comprises administering a therapeutically effective amount of a compound of formula (I) to an individual in need, such as a human. In another aspect, the invention provides a method for treatment as described above, in combination with additional cancer treatments, such as other anti-cancer drugs, surgery, immunotherapy, and/or local therapy, such as radiofrequency ablation. In one aspect, the present invention provides a method for treating primary or secondary liver cancer, which comprises administering a pharmaceutical combination comprising a therapeutically effective amount of a compound of formula I, further comprising one or more selected from the group consisting of chemotherapy Agents, multidrug resistance reversal agents, and biological response modifiers. In one embodiment of this aspect, the other therapeutic agent is a chemotherapeutic agent. In one aspect, the invention provides a compound of formula (I) selected from the following compounds:

Or its pharmaceutically acceptable salts. In addition, the present invention relates to a method for preparing the compound of formula (I), for preparing novel intermediates of the compound of formula (I) and the preparation of such intermediates. When the terms "compound of formula (I)", "compound of the invention" or similar terms are used above and below, it is meant to include compounds of formula (I) and any subgroup of compounds of formula (I), including all possible Stereochemically isomeric forms, pharmaceutically acceptable salts, solvates, quaternary amines and metal complexes. The compound of the present invention can be formulated into various pharmaceutical forms for administration purposes. As suitable compositions, all compositions commonly used for oral administration of drugs can be mentioned. In order to prepare the pharmaceutical composition of the present invention, an effective amount of a specific compound as an active ingredient (in the form of an addition salt or solvate as appropriate) is combined with a pharmaceutically acceptable carrier in a uniform mixture, the carrier may A variety of forms are used, depending on the form of preparation required for administration. These pharmaceutical compositions are preferably in unit dosage form suitable for oral administration. For example, in preparing compositions for oral dosage forms, any common pharmaceutical medium may be used, such as in the case of oral liquid preparations (such as suspensions, syrups, elixirs, emulsions, and solutions), such as water, Glycols, oils, alcohols and the like; or in the case of powders, pills, capsules and lozenges, solid carriers such as starch, sugar, kaolin, lubricants, binders, disintegrants and the like. Lozenges and capsules represent the most advantageous oral dosage unit form because they are easy to administer. In this case, solid pharmaceutical carriers are obviously used. Also included are solid form preparations that are converted to liquid form preparations immediately before use. It is especially suitable to formulate the above-mentioned pharmaceutical composition in unit dosage form for ease of use and uniform dosage. The unit dosage form used herein refers to a physical discrete unit suitable as a unit dose, each unit containing a predetermined amount of active ingredient calculated to produce a desired therapeutic effect and a required pharmaceutical carrier. Examples of such unit dosage forms are lozenges (including indented or coated lozenges), capsules, pills, powder packs, flakes and the like, and multiple portions thereof. In general, the expected daily effective amount of cancer is about 0.01 to about 700 mg/kg body weight, or about 0.5 to about 400 mg/kg body weight, or about 1 to about 250 mg/kg body weight, or about 2 to about 200 mg/kg kg body weight, or about 10 to about 150 mg/kg body weight. It may be appropriate to administer the required dose in two, three, four or more sub-doses at appropriate intervals throughout the day. The sub-dose can be formulated into unit dosage forms, for example, containing about 1 to about 5000 mg, or about 50 to about 3000 mg, or about 100 to about 1000 mg, or about 200 to about 600 mg, or about 100 per unit dosage form To about 400 mg of active ingredient. The compounds of the present invention may only exhibit anticancer effects and/or enhance the ability of another anticancer agent to exhibit anticancer effects. The compounds of the present invention appear as defined stereoisomers. The absolute configuration of these compounds can be determined using methods known in the art, such as X-ray diffraction or NMR and/or inferred from starting materials with known stereochemistry. The pharmaceutical composition according to the invention will preferably comprise a substantially stereoisomerically pure preparation of said stereoisomer. The pure stereoisomeric forms of the compounds and intermediates described herein are defined as other enantiomeric or diastereoisomeric forms that are substantially free of the same basic molecular structure of the compounds or intermediates. In particular, the term "stereomerically pure" relates to a stereoisomer excess of at least 80% (ie, a minimum of 90% of one isomer and a maximum of 10% of other possible isomers) to 100% of the stereo Compounds or intermediates with heterogeneous excess (ie, 100% of one isomer contains no other), more specifically, compounds or intermediates with a stereoisomeric excess of 90% to 100%, or even more specifically , With a stereoisomeric excess of 94% to 100% and most specifically, a stereoisomeric excess of 97% to 100%. The terms "enantiomerically pure" and "diastereomerically pure" should be understood in a similar manner, but with regard to the enantiomeric excess and diastereomeric excess of the mixture in question, respectively. Pure stereoisomeric forms of the compounds and intermediates of the present invention can be obtained by applying procedures known in the art. For example, enantiomers can be separated from each other by selectively crystallizing their diastereoisomeric salts using optically active acids or bases. Examples thereof are tartaric acid, dibenzoyl tartaric acid, xylyl tartaric acid and camphorsulfonic acid. Alternatively, enantiomers can be separated by chromatographic techniques using a palmar stationary phase. Such pure stereochemically isomeric forms can also be obtained from the corresponding pure stereochemically isomeric forms of suitable starting materials, provided that the reaction proceeds in a stereospecific manner. Preferably, if a specific stereoisomer is required, the compound is synthesized by a stereospecific preparation method. These methods preferably use enantiomerically pure starting materials. The diastereomeric racemates of the compounds of the present invention can be obtained by conventional methods alone. Suitable physical separation methods suitable for use are, for example, selective crystallization and chromatography, such as column chromatography. When a phosphorus atom is present in the compound of the present invention, the phosphorus atom may represent an opposite palm center. According to the Cahn-Ingold-Prelog priority rule, the opposition of the center is expressed as "R" or "S". When no palmarity is indicated, it is intended to include R- and S-isomers, and mixtures of both, ie, diastereoisomeric mixtures. In a preferred embodiment of the present invention, it includes a stereoisomer whose phosphorus atom has an S-configuration. These stereoisomers represented as S _P. In other embodiments of the present invention, stereoisomers with phosphorus atoms having an R-configuration are included. These stereoisomers are expressed as R _P. In other embodiments of the present invention, it includes diastereoisomeric mixtures, ie mixtures of compounds with phosphorus atoms having an R- or S-configuration. The present invention also includes isotopically-labeled compounds of formula (I) in which one or more atoms are replaced by an isotope of the atom, that is, an atom having the same atomic number as the atoms commonly found in nature, but different atomic weights. Examples of isotopes that can be incorporated into compounds of formula (I) include, but are not limited to, isotopes of hydrogen, such as ² H and ³ H (also deuterium D and tritium T, respectively); carbon, such as ¹¹ C, ¹³ C, and ¹⁴ C; nitrogen, such as ¹³ N and ¹⁵ N; oxygen, such as ¹⁵ O, ¹⁷ O, and ¹⁸ O; phosphorus, such as ³¹ P and ³² P; sulfur, such as ³⁵ S; fluorine, such as ¹⁸ F; chlorine, such as ³⁶ Cl; Bromine, such as ⁷⁵ Br, ⁷⁶ Br, ⁷⁷ Br, and ⁸² Br; and iodine, such as ¹²³ I, ¹²⁴ I, ¹²⁵ I, and ¹³¹ I. The choice of isotopes included in an isotope-labeled compound will depend on the specific application of the compound. For example, for drug or matrix tissue distribution analysis, compounds in which radioisotopes such as ³ H or ¹⁴ C are incorporated are generally most useful. For radiographic applications, such as positron emission tomography (PET), positron emitting isotopes such as ¹¹ C, ¹⁸ F, ¹³ N, or ¹⁵ O will be useful. Incorporation of heavier isotopes (such as deuterium, ie ² H) can provide the compound of formula (I) with greater metabolic stability, which can, for example, increase the compound's in vivo half-life or reduce the dosage requirements. The isotopically-labeled compounds of the present invention can be substituted for the corresponding non-isotopically-labeled reagents or starting materials by methods similar to those described in the processes and/or examples below, using appropriate isotopically-labeled reagents or starting materials, Or prepared by conventional techniques known to the skilled person. Pharmaceutically acceptable addition salts include the therapeutically active acid and base addition salt forms of compounds of formula (I). Of interest is the free (ie non-salt) form of the compound of formula (I) or a subgroup thereof. Pharmaceutically acceptable acid addition salts are preferably obtained by treating the alkali form with the appropriate acid. Suitable acids include, for example, inorganic acids, such as hydrohalic acids, such as hydrochloric acid or hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and similar acids; or organic acids, such as acetic acid, propionic acid, glycolic acid, lactic acid, pyruvic acid, oxalic acid ( (I.e. oxalic acid), malonic acid, succinic acid (i.e. succinic acid), maleic acid, fumaric acid, malic acid (i.e. hydroxysuccinic acid), tartaric acid, citric acid, methanesulfonic acid, Ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclohexylaminesulfonic acid, salicylic acid, p-aminosalicylic acid, pamoic acid and similar acids. Conversely, these salt forms can be converted to the free base form by suitable base treatment. Compounds of formula (I) containing acidic protons can also be converted into their non-toxic metal or amine addition salt forms by treatment with suitable organic and inorganic bases. Suitable alkali salt forms include, for example, ammonium salts, alkali and alkaline earth metal salts such as lithium, sodium, potassium, magnesium, calcium salts and the like, and salts with organic bases such as benzathine, N-methyl-D-glucose Glycosamine, Hydrazone, and salts with amino acids, such as arginine, lysine, and the like. Some compounds of formula (I) may also exist in their tautomeric forms. For example, the tautomeric form of an amide group (-C(=O)-NH-) is an imino alcohol (-C(OH)=N-), which becomes stable in the aromatic ring . Although the form is not explicitly indicated in the structural formula shown herein, it is intended to be included in the scope of the present invention. The terms and expressions used in the abstract, specification and patent application scope should be understood in accordance with the following definitions unless otherwise indicated. The meaning of each term is independent at each occurrence. These definitions apply regardless of whether the terms are used alone or in combination with other terms, unless otherwise indicated. Terms or expressions used in the text that are not clearly defined should be understood to have their common meanings used in the field. Chemical names, common names, and chemical structures can be used interchangeably to describe the same structure. If a compound is referred to using both a chemical structure and a chemical name and there is an ambiguity between the structure and the name, the structure prevails. "C _m -C _n alkyl" itself or in a compound expression such as C _m -C _n haloalkyl, C _m -C _n alkylcarbonyl, C _m -C _n alkylamine, etc. indicates that it has the specified number of carbon atoms The straight-chain or branched-chain aliphatic hydrocarbon group, for example, C ₁ -C ₄ alkyl means an alkyl group having 1 to 4 carbon atoms. C ₁ -C ₆ alkyl has the corresponding meaning, and also includes all linear and branched chain isomers of pentyl and hexyl. Preferred alkyl groups for use in the present invention are C ₁ -C ₆ alkyl groups, including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, second butyl, third butyl , N-pentyl and n-hexyl, especially C ₁ -C ₄ alkyl, such as methyl, ethyl, n-propyl, isopropyl, third butyl, n-butyl and isobutyl. Methyl and isopropyl are generally preferred. The alkyl group may be unsubstituted or substituted with one or more substituents which may be the same or different, and each substituent is independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyanide Group, hydroxyl, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH ₂ , -NH (alkyl), -N (alkyl) ₂ ,- NH(cycloalkyl), -OC(=O)-alkyl, -OC(=O)-aryl, -OC(=O)-cycloalkyl, -C(=O)OH and -C(= O) O-alkyl. It is generally preferred that the alkyl group is unsubstituted unless otherwise indicated. "C ₂ -C _n alkenyl" means a straight or branched aliphatic hydrocarbon group containing at least one carbon-carbon double bond and having the specified number of carbon atoms, for example, C ₂ -C ₄ alkenyl means having 2 to 4 carbons Atom alkenyl; C ₂ -C ₆ alkenyl means alkenyl having 2 to 6 carbon atoms. Non-limiting alkenyl groups include vinyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl, and hexenyl. The alkenyl group may be unsubstituted or substituted with one or more substituents which may be the same or different, and each substituent is independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyanide Group, hydroxyl, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH ₂ , -NH (alkyl), -N (alkyl) ₂ ,- NH(cycloalkyl), -OC(=O)-alkyl, -OC(=O)-aryl, -OC(=O)-cycloalkyl, -C(=O)OH and -C(= O) O-alkyl. It is generally preferred that the alkenyl group is unsubstituted unless otherwise indicated. "C ₂ -C _n alkynyl" means a straight or branched aliphatic hydrocarbon group containing at least one carbon-carbon triple bond and having the specified number of carbon atoms, for example, C ₂ -C ₄ alkynyl means having 2 to 4 carbon atoms C ₂ -C ₆ alkynyl means an alkynyl group having 2 to 6 carbon atoms. Non-limiting alkynyl groups include ethynyl, propynyl, 2-butynyl and 3-methylbutynyl, pentynyl, and hexynyl. The alkynyl group may be unsubstituted or substituted with one or more substituents which may be the same or different, and each substituent is independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, cyanide Group, hydroxyl, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH ₂ , -NH (alkyl), -N (alkyl) ₂ ,- NH(cycloalkyl), -OC(=O)-alkyl, -OC(=O)-aryl, -OC(=O)-cycloalkyl, -C(=O)OH and -C(= O) O-alkyl. It is generally preferred that the alkynyl group is unsubstituted, unless otherwise indicated. As used herein, the term "C _m -C _n haloalkyl" means that at least one C atom is substituted with halogen (eg, C _m -C _n haloalkyl may contain one to three halogen atoms), preferably chlorine or fluorine. C _m -C _n alkyl. A typical haloalkyl group is a C ₁ -C ₂ haloalkyl group, wherein the halo group preferably represents fluorine. Exemplary haloalkyl groups include fluoromethyl, difluoromethyl, and trifluoromethyl. As used herein, the term "C _m -C _n hydroxyalkyl group" means wherein at least one C atom is substituted with a hydroxyl group of C _m -C _n alkyl. Typical C _m -C _n hydroxyalkyl group in which a C atom is a substituted hydroxy of C _m -C _n alkyl. Exemplary hydroxyalkyl groups include hydroxymethyl and hydroxyethyl. As used herein, the term "C _m -C _n alkyl group" represents a group wherein at least one C atom of the amine group via a group C _m -C _n. Typical C _m -C _n alkyl group wherein one C atom is substituted with one of alkyl group C _m -C _n. Exemplary aminoalkyl groups include aminomethyl and aminoethyl. As used herein, the term "C _m -C _n alkylene" refers to a straight or branched chain divalent alkyl group having the specified number of carbon atoms. The preferred C _m -C _n alkylene group used in the present invention is a C ₁ -C ₃ alkylene group. Non-limiting examples of alkylene include -CH ₂ -, -CH ₂ CH ₂ -, -CH ₂ CH ₂ CH ₂ -, -CH(CH ₃ )CH ₂ CH ₂ -, -CH(CH ₃ )- and- CH(CH(CH ₃ ) ₂ )-. The term "Me" means methyl and "MeO" means nailoxy. The term "C _m -C _n alkylcarbonyl" refers to a group in which the C _m -C _n alkyl portion is as defined above, of the formula C _m -C _n alkyl-C(=O)-. Typically, "C _m -C _n alkylcarbonyl" is C ₁ -C ₆ alkyl-C(=O)-. "C _m -C _n alkoxy" means a group C _m -C _n alkyl-O- in which C _m -C _n alkoxy is as defined above. Of particular interest are C ₁ -C ₄ alkoxy groups, which include methoxy, ethoxy, n-propoxy, isopropoxy, third butoxy, n-butoxy and isobutoxy. Methoxy and isopropoxy are generally preferred. C ₁ -C ₆ alkoxy has the corresponding meaning and extends to all linear and branched chain isomers including pentyloxy and hexyloxy. The term "C _m -C _n alkoxycarbonyl" means a group in which C _m -C _n alkoxy is as defined above and is of the formula C _m -C _n alkoxy-C(=O)-. Typically, "C _m -C _n alkoxycarbonyl" is C ₁ -C ₆ alkoxy-C(=O)-. The term "amino" refers to the group -NH ₂ . The term "halo" means a halogen group such as fluorine, chlorine, bromine or iodine. Typically, the halogen group is fluorine or chlorine. The term "aryl" means phenyl, biphenyl or naphthyl. The term "heterocycloalkyl" refers to a stable saturated monocyclic 3-7 membered ring containing 1-3 heteroatoms independently selected from O, S and N. In one embodiment, the stable saturated monocyclic 3-7 member ring contains 1 heteroatom selected from O, S, and N. In the second embodiment, the stable saturated monocyclic 3-7 membered ring contains 2 heteroatoms independently selected from O, S and N. In the third embodiment, the stable saturated monocyclic 3-7 membered ring contains 3 heteroatoms independently selected from O, S and N. Stable saturated monocyclic 3-7 membered rings containing 1-3 heteroatoms independently selected from O, S and N are generally 5-7 membered rings, such as 5 or 6 membered rings. Heterocycloalkyl may be unsubstituted or substituted with one or more substituents which may be the same or different, and each substituent is independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl , Cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH ₂ , -NH (alkyl), -N (alkyl) ₂ , -NH(cycloalkyl), -OC(=O)-alkyl, -OC(=O)-aryl, -OC(=O)-cycloalkyl, -C(=O)OH and -C (=O)O-alkyl. It is generally preferred that the heterocycloalkyl group is unsubstituted unless otherwise indicated. The term "heteroaryl" refers to a stable monocyclic or bicyclic aromatic ring system containing 1-4 heteroatoms independently selected from O, S, and N, each ring having 5 or 6 ring atoms. In one embodiment of the invention, the stable monocyclic or bicyclic aromatic ring system contains one heteroatom selected from O, S, and N, each ring having 5 or 6 ring atoms. In a second embodiment of the invention, the stable monocyclic or bicyclic aromatic ring system contains two heteroatoms independently selected from O, S and N, each ring having 5 or 6 ring atoms. In a third embodiment, the stable monocyclic or bicyclic aromatic ring system contains three heteroatoms independently selected from O, S, and N, each ring having 5 or 6 ring atoms. In a fourth embodiment, the stable monocyclic or bicyclic aromatic ring system contains four heteroatoms independently selected from O, S and N, each ring having 5 or 6 ring atoms. An example of a heteroaryl group includes flavonoids. The term "C ₃ -C _n cycloalkyl" means a cyclic monovalent alkyl group having a specified number of carbon atoms, for example, C ₃ -C ₇ cycloalkyl means a cyclic monovalent alkyl group having 3 to 7 carbon atoms. The preferred cycloalkyl groups used in the present invention are C ₃ -C ₄ alkyl groups, namely cyclopropyl and cyclobutyl. The cycloalkyl group may be unsubstituted or substituted with one or more substituents which may be the same or different, and each substituent is independently selected from the group consisting of halo, alkenyl, alkynyl, aryl, cycloalkyl, Cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH ₂ , -NH (alkyl), -N (alkyl) ₂ , -NH(cycloalkyl), -OC(=O)-alkyl, -OC(=O)-aryl, -OC(=O)-cycloalkyl, -C(=O)OH and -C( =O)O-alkyl. It is generally preferred that the cycloalkyl group is unsubstituted unless otherwise indicated. The term "alkyl group C _m -C _n" represents the amino substituted by C _m -C _n alkyl group as defined above it, i.e., the alkyl group of a hydrogen atom is NH ₂ - a displaceable group. Typically, "amino C _m -C _n alkyl" is an amine C ₁ -C ₆ alkyl. The term "C _m -C _n alkyl amino carbonyl" denotes C _m -C _n alkyl group as defined above, of a carbonyl group, wherein the alkyl group of a hydrogen atom is NH ₂ - a displaceable group. Typically, "amino C _m -C _n alkylcarbonyl" is an amine C ₁ -C ₆ alkylcarbonyl. Examples of the amino group C _m -C _n alkylcarbonyl include, but are not limited to, glycinyl group: C(=O)CH ₂ NH ₂ , propylamine group: C(=O)CH(NH ₂ )CH ₃ , valamine Acyl group: C=OCH(NH ₂ )CH(CH ₃ ) ₂ , white amine acetyl group: C(=O)CH(NH ₂ )(CH ₂ ) ₃ CH ₃ , iso white amine acetyl group: C(=O ) CH(NH ₂ )CH(CH ₃ )(CH ₂ CH ₃ ) and n-leucine acid group: C(=O)CH(NH ₂ )(CH ₂ ) ₃ CH ₃ and its similar groups. This definition is not limited to naturally occurring amino acids. As used herein, the term "(=O)" when attached to a carbon atom forms a carbonyl moiety. It should be noted that when the valence allows, one atom can only carry one pendant oxygen group. The terms "monophosphate, diphosphate and triphosphate" refer to the following groups:

and

. As used herein, the position of the group on any molecular part used in the definition can be anywhere on that part, mainly because it is chemically stable. When there are any variables that appear multiple times in any part, each definition is independent. The term "solvate" encompasses any pharmaceutically acceptable solvate that the compound of formula (I) and its salts can form. The solvates are, for example, hydrates, alcoholates, such as ethanolates, propanolates and the like, especially hydrates. As used herein, the term "prodrug" refers to a drug precursor that is easily transformed in the body through metabolism and/or chemical processes to obtain an active compound after administration to an individual. As used in the text, the expression "liver-targeted prodrug" means a prodrug that is metabolized into its active substance mainly in the liver. As used herein, the expression "liver cancer" is intended to include primary and secondary liver cancer, that is, liver metastases from cancers derived from the liver and cancers from other organs, respectively. Related terms should be understood in conjunction with the definitions provided above and common uses in the technical field. In general, the names of the compounds used in this application were generated using ChemDraw Ultra 12.0. In addition, if the stereochemistry of a structure or a part of a structure is not indicated by, for example, bold or dashed lines, the structure or a part of the structure should be understood to cover all stereoisomers thereof. General Synthesis Methods The present invention can be prepared by a variety of methods, such as those shown below and described in the illustrative synthetic schemes. All starting materials and reagents can be purchased from suppliers or can be prepared according to the literature procedures described in the references using methods well known to those skilled in the art. Scheme 1 illustrates the general route of the compound of formula (I).

Scheme 1 The commercially available trasatabine derivative (1a) prepared as described above and the desired aminophosphate reagent (1b) (where Lg is a suitable leaving group, such as halogen (such as chlorine) or activated phenol ( Such as pentachlorophenol, p-nitrophenol, pentafluorophenol or the like)) in an inert solvent (such as ether, such as ether or THF), or halogenated hydrocarbons (such as dichloromethane), in a base (such as N-methyl Condensation in the presence of imidazole (NMI)) or Grignard reagent (such as tertiary butylmagnesium chloride or the like) gives an amino phosphate derivative (1c). Aminophosphate reagent (1b) used in the above process (where Lg is chlorine, that is, chlorinated aminophosphate) can be prepared in two steps as described in Scheme 2 using phosphorus oxychloride (POCl ₃ ) as the starting material .

Scheme 2 POCl ₃ is condensed with the desired alcohol R ¹³ OH in an inert solvent (such as Et ₂ O) to obtain an alkoxy or aryloxy dichloride phosphate (2a). Amino acid derivative followed by reaction with (2b) wherein R ^{3 'is} H of chloramine-yl phosphate (2c). If necessary, the obtained chloramine phosphoric acid ester (2c) can be converted into the corresponding phosphorylating agent with active phenol as the leaving group (such as pentafluorophenol or p-NO ₂ -phenol) as outlined in Scheme 3. .

Scheme 3 This conversion is preferably carried out by reacting the chlorine derivative (2c) with the desired active phenol in the presence of a base (such as triethylamine or the like) to provide phosphorylating agents (3a) and (3b). The use of various protective groups (PG) for the above process is known to those skilled in the art, and its applications and alternatives are detailed in the literature, see eg Greene TW, Wuts PGM Protective groups in organic synthesis, Second Edition New York: Wiley; 1995. As used herein, the term "N-protecting group" or "N-protected" refers to one intended to protect the N-terminus of an amino acid or peptide or protect the amino acid from undesired reactions during synthetic procedures And other groups. Commonly used N-protecting groups are disclosed in Greene. N-protecting groups include acetyl groups, such as methyl acetyl, acetyl propyl, propyl acetyl, tert-pentyl acetyl, tertiary butyl acetyl acetyl, 2-chloroethyl acetyl, 2-bromo acetyl acetyl, tri Fluoroethylenyl, trichloroethylenyl, o-xylylene, o-nitrophenoxyethylenyl, α-chlorobutyryl, benzoyl, 4-chlorobenzyl, 4-bromobenzene Formyl, 4-nitrobenzyl and similar groups; sulfonyl, such as benzenesulfonyl, p-toluenesulfonyl and similar groups; carbamate-forming groups, such as benzyl Oxycarbonyl, p-chlorobenzyloxy-carbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3,4-di Methoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl, 3,4,5-trimethoxybenzyloxycarbonyl, 1-(p-biphenyl)-1-methylethoxycarbonyl, α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, dibenzyloxycarbonyl, third butoxy Carbonyl, diisopropylmethoxycarbonyl, isopropoxycarbonyl, ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl, 2,2,2-trichloroethoxycarbonyl, phenoxycarbonyl , 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl, adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and the like; alkyl , Such as benzyl, triphenylmethyl, benzyloxymethyl and similar groups; and silane groups, such as trimethylsilyl and similar groups. Preferred N-protecting groups include methyl acetyl, ethyl acetyl, benzoyl acetyl, tert-pentyl acetyl, third butyl acetyl acetyl, phenyl sulfonyl acetyl, benzyl (Bz), third butyloxy Carbonyl (BOC) and benzyloxycarbonyl (Cbz). Hydroxy and/or carboxyl protecting groups are also detailed above in Greene and include ethers such as methyl ether; substituted methyl ethers such as methoxymethyl, methylthiomethyl, benzyloxymethyl, Third butoxymethyl, 2-methoxyethoxymethyl and similar groups; silane ethers, such as trimethylsilyl (TMS), third butyldimethylsilyl (TBDMS) , Tribenzylsilyl, triphenylsilyl, tert-butyldiphenylsilyl, triisopropylsilyl and similar groups; substituted ethyl ethers, such as 1-ethoxymethyl, 1-methyl-1-methoxyethyl, tertiary butyl, allyl, benzyl, p-methoxybenzyl, diphenylmethyl, triphenylmethyl and similar groups; aromatic Alkyl groups such as trityl and pixyl (9-hydroxy-9-phenyldibenzopiperan derivatives, especially chloride). Ester hydroxy protecting groups include esters, such as formate, benzyl formate, chloroacetate, methoxyacetate, phenoxyacetate, pivalate, adamantate, oleate (mesitoate), parabens and their analogs. Carbonate hydroxyl protecting groups include methylvinyl, allyl, cinnamyl, benzyl and similar groups.

因此，本發明之多種實施例及中間物現將藉由以下實例說明。該等實例僅用於進一步說明本發明並不以任何方式限制本發明之範疇。化合物名稱係藉由ChemDraw Ultra軟體, Cambridgesoft, 12.0.2版產生。除了以上定義，以下縮寫用於以上合成流程及以下實例中。若文中所用之縮寫未定義，則其具有其公認含義。

曲沙他濱之製備

步驟 1) ((2,2- 二甲氧基乙氧基 ) 甲基 ) 苯 (Tr-1) 在0℃下，將苄基溴(56.03 mL，0.471 mol)及NaOH(20.7 g，0.518 mol)添加至2,2-二甲氧基乙醇(50 g，0.471 mol)於DMF(200 mL)中之攪拌溶液中並在室溫下攪拌反應混合物16小時。反應完全後(TLC)，添加飽和氯化鈉溶液(500 mL)並利用DCM(1 L)萃取反應混合物，使有機相乾燥(Na₂ SO₄ )並濃縮且所得粗產物藉由矽膠管柱層析在60-120二氧化矽上利用4-6% EtOAc/己烷純化以得到呈液體狀之標題化合物(60 g，60%)。步驟 2) (5S)-5-((4S)-2-(( 苄氧基 ) 甲基 )-1,3- 二氧雜環戊烷 -4- 基 )-3,4- 二羥基呋喃 -2(5H)- 酮 (Tr-2) 將L-抗壞血酸(44.9 g，0.255 mol)添加至化合物Tr-1(60 g，0.306 mol)於無水乙腈(898 mL)中之溶液中，隨後添加pTSA單水合物(15.5 g，0.076 mol)並將反應混合物在90℃下加熱1小時。反應完全後(TLC)，蒸餾掉一半體積之乙腈並重複該過程兩次。完全去除溶劑並得到呈立體異構體混合物形式之標題化合物(91 g)。產物不作進一步純化直接用於下一步驟。步驟 3) (2R)-2-((4S)-2-(( 苄氧基 ) 甲基 )-1,3- 二氧雜環戊烷 -4- 基 )-2- 羥基乙酸 (Tr-3) 在室溫下，將化合物Tr-2(91.7 g，0.297 mol)添加至K₂ CO₃ (86.3 g，0.625 mol)於H₂ O(509 mL)中之攪拌溶液中。緩慢添加H₂ O₂ (80 mL，0.71 mol，30% v/v)並將溶液冷卻至0℃並隨後攪拌24小時。減壓下移除溶劑，添加EtOH(100 mL)並使混合物在迴流下加熱30分鐘，隨後過濾。添加EtOH(100 mL)以得到固體殘餘物並使混合物在迴流下加熱30分鐘(兩次)。使所收集之過濾物在真空下濃縮，得到呈固體狀之標題化合物(90 g)。步驟 4) (2S,4S)-2-(( 苯氧基 ) 甲基 )-1,3- 二氧雜環戊烷 -4- 甲酸 (Tr-4a) 及 (2R,4S)-2-(( 苯氧基 ) 甲基 )-1,3- 二氧雜環戊烷 -4- 甲酸 (Tr-4b) 在30分鐘內，將次氯酸鈉(650 ml，0.881 mol，9-10%水溶液)逐滴添加至化合物Tr-3(90 g，0.294 mol)及RuCl3,xH₂ O(1.22 g，0.0058 mol)在水(ml pH=8室溫)中之劇烈攪拌溶液中。藉由添加1 M NaOH溶液使pH保持在8。室溫下攪拌該反應混合物3小時，隨後在35℃下加熱12小時。反應完全後(TLC)，在0℃下，將1.5 N HCl添加至該反應混合物中直至達到pH 6，隨後添加EtOAc(1 L)。利用鹽水(2×100 mL)洗滌有機相，乾燥(Na₂ SO₄ )，過濾並濃縮。所得粗產物藉由矽膠管柱層析在230-400二氧化矽上利用20% EtOAc/石油醚純化以得到呈異構體混合物形式之化合物4a+4b。隨後，異構體藉由管柱層析在二氧化矽230-400上利用0.9% MeOH/DCM及0.1% AcOH作為溶離劑加以分離以得到2R異構體(20 g，28%)。步驟 5) 乙酸 (2S)-2-(( 苄氧基 ) 甲基 )-1,3- 二氧雜環戊烷 -4- 基酯 (Tr-5) 將吡啶(13.2 ml)及乙酸鉛(79.8 g，0.180 mol)添加至化合物Tr-4a(33 g，0138 mol)於乙腈(660 mL)中之溶液中並使該混合物在室溫下攪拌16小時。反應完全後(TLC)，過濾反應混合物，濃縮濾液並將殘餘物溶於EtOAc(500 mL)中，用水(100 mL)及飽和氯化鈉溶液(100 mL)洗滌並經Na₂ SO₄ 乾燥。移除溶劑後，粗產物藉由管柱層析在60-120二氧化矽上利用12-15% EtOAc/石油醚梯度加以純化以得到呈液體狀之標題化合物(16 g，47%)。步驟 6) 乙酸 (2S)-2-( 羥甲基 )-1,3- 二氧雜環戊烷 -4- 基酯 (Tr-6) 將Pd/C(3.2 g，20% w/w)添加至化合物Tr-5(16 g)在無水甲醇(160 mL)中之攪拌溶液中，使該反應混合物氫化3小時。反應完全後(TLC)，藉由矽藻土過濾反應混合物。在減壓下濃縮濾液並將所得粗製之標題化合物(10 g，97%)直接用於下一步驟。步驟 7) 乙酸 ((2S)-4- 乙醯氧基 -1,3- 二氧雜環戊烷 -2- 基 ) 甲酯 (Tr-7) 在0℃下，將乙酸酐(8.22 ml，0.080 mol)添加至化合物Tr-6(5.74 g，0.0354 mol)在吡啶(107 ml)中之攪拌溶液中並使該反應混合物在室溫下攪拌16小時。反應完全後(TLC)，利用稀HCl(10 mL)淬滅該反應混合物並萃取至EtOAc(100 mL)中。分離有機相，乾燥(Na₂ SO₄ )、過濾並濃縮。所得粗產物藉由管柱層析在230-400二氧化矽上利用10-15% EtOAc/石油醚梯度溶離加以純化以得到呈液體狀之標題化合物(4.97 g，68%)。步驟 8) 乙酸 ((2S,4S)-4-(4-( 苄胺基 )-2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲酯 (Tr-8a) 使N-苯甲醯基胞嘧啶(12.1 g，56.3mmol)、硫酸銨(催化量)及六甲基二矽氮烷(HMDS)(67.4 ml，418 mmol)之混合物迴流1小時。在40℃下，減壓下移除HMDS並將殘餘物溶於無水1,2-二氯乙烷(57 ml)中並添加化合物Tr-7(5.7 g，27.9 mmol)在無水1,2-二氯乙烷(57 ml)中之溶液，隨後逐滴添加TMSOTf(10.2 ml，45.7 mmol)。使該反應混合物在室溫下攪拌1小時，隨後添加NaHCO₃ 水溶液並使該混合物攪拌30分鐘。藉由矽藻土過濾出所得固體並將濾液溶於EtOAc(200 mL)中，利用水(50 mL)洗滌並乾燥(Na₂ SO₄ )。減壓下移除溶劑後，粗產物藉由管柱層析在230-400二氧化矽上利用10-15% EtOAc/石油醚梯度加以純化以得到變旋異構體之混合物，該變旋異構體之混合物藉由SFC純化進一步分離以得到呈白色固體狀之標題化合物(3 g，30%)。步驟 9) 4- 胺基 -1-((2S,4S)-2-( 羥甲基 )-1,3- 二氧雜環戊烷 -4- 基 ) 嘧啶 -2(1H)- 酮 (Tr-9) 在室溫下，使化合物Tr-8a(3 g)、飽和甲醇氨溶液(180 ml)之混合物在密封管中攪拌16小時。反應完全後(TLC)，減壓下移除溶劑且粗產物藉由管柱層析在230-400二氧化矽上利用10-13% MeOH/DCM梯度溶離加以純化，得到呈固體狀之標題化合物(1.5 g，85%)。¹ H NMR 400 MHz DMSO-d₆ δ: 3.63-3.65 (2H), 4.04-4.07 (2H), 4.92-4.94 (1H), 5.18-5.21 (1H), 5.72-5.74 (1H), 6.16-6.18 (1H), 7.14 (1H), 7.26 (1H), 7.80-7.82 (1H)。5-F- 曲沙他濱之製備

步驟 1) 苯甲酸 ((2S,4R)-4-(4- 苯甲醯胺基 -5- 氟 -2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲酯 (5-F-Tr-1a) 及 苯甲酸 ((2S,4S)-4-(4- 苯甲醯胺基 -5- 氟 -2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲酯 (5-F-Tr-1b) 使5-氟苯甲醯基胞嘧啶(9.1 g，39.5 mmol)、硫酸銨(催化量)及六甲基二矽氮烷(140 ml)之混合物迴流14小時。在40℃下，減壓下移除HMDS並將殘餘物溶於無水1,2-二氯乙烷(50 ml)中並添加化合物苯甲酸((2S)-4-乙醯氧基-1,3-二氧雜環戊烷-2-基)甲酯(7 g，26.30 mmol)在無水1,2-二氯乙烷(50 ml)中之溶液隨後逐滴添加TMS-OTf(11.6 g，52.6 mmol)。使該反應混合物在室溫下攪拌2小時，隨後將NaHCO₃ 水溶液添加至反應混合物中並再將該混合物攪拌30分鐘。藉由矽藻土過濾所得固體並將濾液溶於EtOAc(500 mL)中，利用水(50 mL)洗滌並乾燥(Na₂ SO₄ )。減壓下移除溶劑且粗產物藉由管柱層析在230-400二氧化矽上利用50-60% EtOAc/石油醚梯度加以純化以得到呈固體狀之純標題化合物(1.7g，18%)。步驟 2) 4- 胺基 -5- 氟 -1-((2S,4S)-2-( 羥甲基 )-1,3- 二氧雜環戊烷 -4- 基 ) 嘧啶 -2(1H)- 酮 (5-F-Tr) 在室溫下，使化合物5-F-Tr-1b(1.7 g)、飽和甲醇氨溶液(34 ml)之混合物在密封管中攪拌16小時，隨後減壓下移除溶劑且粗產物藉由管柱層析在230-400二氧化矽上利用5% MeOH/DCM梯度加以純化以得到呈固體狀之標題化合物(0.8 g，68%)。製備以下苯酚並用於製備本發明化合物之中間物。苯酚 1

步驟 a) 1-(3-(( 第三丁基二甲基矽烷基 ) 氧基 ) 苯基 ) 乙酮 (Ph1-a) 將咪唑(4.46 g，65.5 mmol)添加至3-羥基苯乙酮(4.46 g，32.8 mmol)在DMF(6 mL)中之溶液中。5分鐘後，添加TBDMS-Cl (4.69 g，31.1 mmol)於DMF(4 mL)中之溶液。使該反應混合物在室溫下攪拌90分鐘，隨後傾入含有5% EtOAc之己烷(200 mL)中並利用1M HCl (60 mL)、水(60 mL)、飽和碳酸氫鈉(2×60 mL)、水(60 mL)及鹽水(60 mL)洗滌。有機層經Na₂ SO₄ 乾燥，過濾並濃縮且所得殘餘物藉由矽膠快速層析利用己烷/EtOAc溶離加以純化以得到標題化合物(5.7 g，69%)。步驟 b) 第三丁基二甲基 (3-( 丙 -1- 烯 -2- 基 ) 苯氧基 ) 矽烷 (Ph1-b) 在氮氣下，將甲基(三苯基鏻)溴化物(10.2 g，28.4 mmol)懸浮在無水THF(30 mL)中並將懸浮液冷卻至0℃。將正丁基鋰(17.8 mL，28.4 mmol)逐滴添加至該混合物中並在室溫下使所得溶液攪拌30分鐘。將Ph1-a(5.7 g，22.8 mmol)添加至該混合物中並使反應在室溫下繼續進行60分鐘。利用碳酸氫鈉水溶液淬滅反應並利用乙醚(50 mL)萃取。利用碳酸氫鈉溶液洗滌有機層，乾燥(Na₂ SO₄ )，過濾並濃縮。所得殘餘物藉由矽膠塞利用己烷溶離加以純化以得到標題化合物(3.9 g，69%)。步驟 c) 第三丁基二甲基 (3-(1- 甲基環丙基 ) 苯氧基 ) 矽烷 (Ph1-c) 在氮氣下，在10分鐘內，將二乙基鋅之己烷溶液(439.2 mmol)逐滴添加至烯烴Ph1-b(3.9 g，15.7 mmol)在1,2-二氯乙烷(60 mL)中之冷卻(0℃)溶液中。逐滴添加二碘甲烷(6.32 mL，78.5 mmol)並使所得混合物在0℃下攪拌30分鐘隨後使其達到室溫隔夜。將該混合物傾入氯化銨之冰冷溶液中並利用乙醚萃取。利用飽和碳酸氫鈉洗滌有機層，乾燥(Na₂ SO₄ )，過濾並濃縮。將粗產物溶於己烷中並將剩餘的二碘甲烷丟棄。將己烷層濃縮為粗產物，該粗產物未作進一步純化即用於下一步驟。步驟 d) 3-(1- 甲基環丙基 ) 苯酚 ( 苯酚 1) 將Ph1-c(3.45 g，13.1 mmol)溶於氟化四丁基銨在THF(20 mL，20 mmol)中之1 M溶液中並使所得溶液在室溫下攪拌隔夜。利用1M HCl(50 mL)淬滅反應並利用乙酸乙酯(100 mL)萃取。利用鹽水(2×50 mL)洗滌有機層，乾燥(Na₂ SO₄ )，過濾並濃縮。殘餘物藉由矽膠快速層析利用2-丙醇、EtOAc及己烷之混合物溶離加以純化以得到標題化合物(0.56 g，29%)。MS 147.1 [M-H]^- 。苯酚 2

標題化合物係由4-羥基苯乙酮(6.0 g，44.1 mmol)利用苯酚1之製備所述之方法製備。產率為53%。苯酚 3

步驟 a) 1-(3-( 苄氧基 ) 苯基 ) 環戊醇 (Ph3-a) 將利用鎂升溫之碘添加至鎂屑(1.29 g，52.8 mmol)在無水THF(50 mL)中之懸浮液中。使該混合物迴流並添加約5%之3-溴苯酚(13.9 g，52.8 mmol)溶液。當反應開始時，逐滴添加溴化物溶液並隨後使該混合物迴流一小時以上。將該混合物冷卻至約5℃並逐滴添加環戊酮(4.44 g，52.8 mmol)於THF(50 mL)中之溶液。使該混合物在室溫下攪拌72小時，隨後利用冷卻的飽和氯化銨溶液淬滅反應並利用乙醚(×3)萃取。利用鹽水洗滌有機相，乾燥(Na₂ SO₄ )，過濾並濃縮。所得產物藉由矽膠層析(異己烷/EtOAc)加以純化以得到標題化合物(8.5 g，54%)。步驟 b) 1-( 苄氧基 )-3-( 環戊 -1- 烯 -1- 基 ) 苯 (Ph3-b) 將對甲苯磺酸添加至Ph3-a(8.4 g，28.2 mmol)於苯(100 mL)中之溶液中。用DMF阱使該混合物迴流3小時，隨後冷卻至rt，利用乙醚稀釋並利用飽和碳酸氫鈉溶液及鹽水洗滌。乾燥有機相(Na₂ SO₄ )，過濾並濃縮。產物藉由矽膠層析(異己烷/EtOAc)加以純化以得到標題化合物(6.45 g，91%)。MS 249.4 [M-H]^- 。步驟 c) 3- 環戊基苯酚 ( 苯酚 3) 在22℃及40 PSI下，在10% Pd/碳(1.5 g)存在下，在帕爾裝置(Parr)中使Ph3-b(6.4 g，26 mmol)在EtOAc (75 mL)及EtOH(75 mL)中之溶液氫化隔夜。過濾出催化劑並利用EtOAc及EtOH洗滌。減壓下蒸發溶劑且產物藉由矽膠層析(異己烷/EtOAc)加以純化以得到標題化合物(3.6 g，82%)。MS 161.2 [M-H]^- 。苯酚 4

步驟 a) 第三丁基 (3- 環丙基苯氧基 ) 二甲基矽烷 (Ph4-a) 在110℃下，使(3-溴苯氧基)(第三丁基)二甲基矽烷(5.46 g，19 mmol)、環丙基

酸(2.12 g，24.7 mmol)、磷酸三鉀(14.1 g，66.5 mmol)、三環己基膦(0.53 g，1.9 mmol)以及Pd(OAc)₂ (0.21 g，0.95 mmol)在甲苯(80 mL)及水(4 mL)中之懸浮液攪拌隔夜。利用乙醚稀釋漿液並利用水及鹽水洗滌。乾燥有機相(MgSO₄ )，乾燥並濃縮。粗產物藉由快速管柱層析(EtOAc/己烷)加以純化以得到標題化合物(1.94 g，41%)。步驟 b) 3- 環丙基苯酚 ( 苯酚 4) 將1 M氟化四丁基銨(10.1 ml，10.1 mmol)添加至Ph4-a(1.94 g，7,81 mmol)於THF(25 ml)中之溶液中。使該溶液攪拌2小時，隨後蒸發溶劑並將殘餘物溶於EtOAc中並利用濃NH₄ Cl(水性)洗滌兩次並利用鹽水洗滌一次。乾燥有機相(MgSO₄ )，過濾並濃縮。粗產物藉由快速管柱層析(己烷/乙酸乙酯9：1，含有1%異丙醇)加以純化以得到稍微不純之標題化合物(1.24 g，119%)。苯酚 5

步驟 a) 2-(4- 溴苯氧基 ) 四氫 -2H- 哌喃 (Ph5-a) 將4-溴苯酚(3.75 g，21.7 mmol)溶於3,4-二氫-2H-哌喃(16 ml，175 mmol)中，添加催化量之對甲苯磺酸(15 mg，0.09 mmol)並在22℃下，使該混合物攪拌45分鐘。利用乙醚稀釋該混合物並利用1 M NaOH (水溶液)×2、水洗滌，乾燥(Na₂ SO₄ )並濃縮，得到標題化合物(5.57 g，99%)。步驟 b) 2-(4- 環丙基苯氧基 ) 四氫 -2H- 哌喃 (Ph5-b) 在15分鐘內，將0.5 M溴化環丙基鎂之THF溶液(6.5 ml，3.25 mmol)添加至Ph5-a(552.5 mg，2.15 mmol)、ZnBr(144 mg，0.64 mmol)、三第三丁基膦四氟硼酸鹽(35.6 mg，0.12 mmol)及Pd(OAc)₂ (29.5 mg，0.13 mmol)於THF(4 ml)中之溶液中。在22℃下，將該混合物攪拌90分鐘，隨後在冰浴上冷卻並添加冰水(10 ml)。利用EtOAc×3萃取混合物並利用鹽水洗滌萃取物，隨後乾燥(Na₂ SO₄ )，過濾並濃縮。殘餘物藉由矽膠管柱層析(石油醚/EtOAc)加以純化以得到標題化合物(292 mg，62%)。步驟 c) 4- 環丙基苯酚 ( 苯酚 5) 將對甲苯磺酸單水合物(18.9 mg，0.1 mmol)添加至Ph5-b(2.28 g，10.45 mmol)於MeOH(15 ml)中之溶液中。在120℃下，使該混合物在微波反應器中加熱5分鐘，隨後濃縮並藉由矽膠管柱層析(石油醚/EtOAc)純化。所得固體自石油醚中結晶，得到標題化合物(1.08 g，77%)。苯酚 6

步驟 a) 1-(3- 甲氧基苯基 ) 環丁醇 (Ph6-a) 在0℃與10℃之間，將1 M 溴化3-甲氧基苯基鎂之THF溶液(2.11 g，99.8 mmol)逐滴添加至環丁酮(6.66 g，95 mmol)於乙醚(65 mL)中之攪拌溶液中。使該混合物在0-10℃下攪拌3小時，隨後將該混合物添加至冰冷飽和NH₄ Cl溶液(300 mL)及水(300 mL)中。使該混合物攪拌10分鐘，隨後利用乙醚萃取三次。乾燥有機相(Na₂ SO₄ )，過濾並濃縮。所得粗產物藉由矽膠層析(異己烷/EtOAc)進行純化以得到標題化合物(16.9 g，86%)。步驟 b) 1- 環丁基 -3- 甲氧基苯 (Ph6-b) 將10% Pd/碳(2.5 g)添加至Ph6-a(15.4 g，86.1 mmol)於乙醇(200 mL)中之溶液中，並使該混合物在帕爾裝置中在60 psi下氫化。18小時後，添加另外10% pd/碳(1.5 g)並使該混合物在60 psi下再氫化18小時。過濾出催化劑並利用EtOH及EtOAc洗滌。減壓下濃縮溶液，並粗產物藉由矽膠層析(異己烷/EtOAc)加以分離以得到標題化合物(14.0 g，77%)。步驟 c) 3- 環丁基苯酚 ( 苯酚 6) 在0℃下，將1 M三溴化硼(18.1 g，72.2 mmol)之DCM溶液逐滴添加至Ph6-b(10.6 g，65.6 mmol)於無水DCM(65 mL)中之溶液中。使該混合物在-5℃下攪拌2.5小時，隨後利用冷卻飽和NH₄ Cl溶液淬滅反應並利用DCM萃取三次。乾燥有機相(Na₂ SO₄ )，過濾並濃縮。所得粗產物藉由矽膠層析(異己烷/EtOAc)進行純化以得到標題化合物(9.73 g，88%)。苯酚 7

步驟 a) 1-(4-( 苄氧基 ) 苯基 ) 環丁醇 (Ph7-a) 在迴流下，在約1 h內，將1-(苄氧基)-4-溴苯(2.63 g，100 mmol)於乙醚:THF 1:1 (100 mL)中之溶液逐滴添加至鎂屑(2.43 g)及痕量碘在乙醚(50 mL)中之懸浮液中。添加完成時，使該混合物迴流四小時，隨後冷卻至約0℃。添加無水THF(50 ml)，隨後緩慢添加環丁酮(7.01 g，100 mmol)於乙醚(50 mL)中之溶液並使該混合物達到室溫。攪拌兩小時後，添加冷卻飽和NH₄ Cl溶液(500 ml)並攪拌該混合物15分鐘，隨後利用EtOAc萃取兩次。利用鹽水洗滌有機相，利用硫酸鈉乾燥並在減壓下蒸發。產物藉由矽膠管柱層析加以純化以得到標題化合物(12.5 g，42%)。步驟 b) 4- 環丁基苯酚 ( 苯酚 7) 在氬氣下，將10% Pd/碳(2.55 g，21.5 mmol)添加至Ph7-a(12.4 g，41.4 mmol)於無水EtOH(110 mL)中之溶液中並在45 psi下，在室溫下，使該混合物氫化18小時。過濾出催化劑，利用乙醇洗滌並使溶液濃縮。該產物藉由矽膠層析(異己烷- EtOAc)純化。合併適宜溶離份並濃縮且殘餘物自石油醚中結晶以得到標題化合物(3.15 g，51%)。苯酚 8

4-(1- 甲基環戊基 ) 苯酚 ( 苯酚 8) 在30分鐘內，將1-甲基環戊醇(2.00 g，20.0 mmol)及苯酚(2.07 g，22.0 mmol)在戊烷(50 mL)中之溶液逐滴添加至新鮮AlCl₃ (1.33 g，10 mmol)在戊烷(100 mL)中之懸浮液中。在N₂ 下，在室溫下，將所得混合物攪拌72小時，隨後將反應混合物傾入水/冰及HCl(12 M，20 mmol，1.66 mL)中。利用水(50 mL)及鹽水(50 mL)洗滌有機相，乾燥(Na₂ SO₄ )，過濾並濃縮。粗產物藉由矽膠管柱層析(MeOH – DCM)加以純化以得到標題化合物(426 mg，12%)。苯酚 9

步驟 a) 2-(4- 溴 -3- 甲基苯氧基 ) 四氫 -2H- 哌喃 (Ph9-a) 將pTs(16 mg，0.086 mmol)添加至4-溴-3-甲基苯酚(4.0 g，21.4 mmol)於3,4-二氫-2-H-哌喃(16 mL，175 mmol)中之溶液。使該反應混合物在室溫下攪拌1小時，隨後利用乙醚稀釋並利用1M NaOH(水溶液)及水洗滌。乾燥有機相(Na₂ SO₄ )，過濾及濃縮。粗產物藉由矽膠管柱層析(EtOAc/庚烷)進行純化以產生標題化合物(3.32 g，57%)。步驟 b) 2-(4- 環丙基 -3- 甲基苯氧基 ) 四氫 -2H- 哌喃 (Ph9-b) 將Ph9-a(3.12 g，11.5 mmol)、ZnBr₂ (2.59 g，11.5 mmol)、三第三丁基膦四氟硼酸鹽(0.2 g，0.69 mmol)及Pd(OAc)₂ (258 mg，1.15 mmol)置於燒瓶中並利用N₂ 沖洗燒瓶數次。在攪拌下添加THF(10 mL)，隨後在5分鐘內逐滴添加0.5 M溴化環丙基鎂之THF溶液(35 mL，17.4 mmol)。在室溫下攪拌該混合物，隨後經矽藻土塞過濾，利用MeOH溶離。濃縮溶液且粗產物藉由矽膠管柱層析(EtOAc/庚烷)進行純化以得到標題化合物(1.69 g，57%)。步驟 c) 4- 環丙基 -3- 甲基苯酚 ( 苯酚 9) 將Ph9-b(1.70 g，7.30 mmol)溶於MeOH(20 ml)中並添加pTsxH₂ O(318 mg，1.67 mmol)。使該混合物在22℃下攪拌30分鐘，隨後濃縮。粗產物藉由管柱層析(EtOAc/庚烷)進行純化以產生標題化合物(704 mg，65%)。苯酚 10

步驟 a) 4- 環丙基 -1- 甲氧基 -2- 甲基苯 (Ph10-a) 根據Ph9步驟b中所述程序，使4-溴-1-甲氧基-2-甲基苯(4.39 g，21.9 mmol)與溴化環丙基鎂反應，得到標題化合物(1.54 g，43%)。步驟 b) 4- 環丙基 -2- 甲基苯酚 ( 苯酚 10) 在N₂ 下，在0℃下，將BBr₃ (5 mL，5 mmol)添加至Ph10-a(1.54 g，9.49 mmol)於DCM(7.5 mL)中之溶液中。使該反應物攪拌2小時，隨後利用MeOH(3 mL)淬滅並濃縮。將粗產物溶於EtOAc中並利用鹽水洗滌。乾燥有機相(Na₂ SO₄ )，過濾並濃縮。粗產物藉由矽膠管柱層析加以純化以得到標題化合物(826 mg，59%)。MS 147.11 [M-H]^- 。苯酚 11

4- 環丙基 -3- 甲氧基苯酚 ( 苯酚 11) 標題化合物係根據苯酚9之製備所述之程序由4-溴-3-甲氧基苯酚(1.11 g，5.49 mmol)製備。產率為40%。苯酚 12

步驟 a) 3-( 二甲胺基 )-1-(3- 羥基苯基 ) 丙 -1- 酮 (Ph12-a) 將幾滴HCl添加至3-羥基苯乙酮(4.08 g，30 mmol)、多聚甲醛(4.05 g，45 mmol)及二甲胺鹽酸鹽(2.69 g，33 mmol)在無水EtOH(100 mL)中之溶液中並使該反應混合物迴流18小時。另添加二甲胺鹽酸鹽(0.55當量，1.22 g)、多聚甲醛(0.5當量，1.35 g)及HCl(0.5 mL)並使該反應混合物再迴流4小時，隨後冷卻至室溫。收集沈積之白色固體並利用冷EtOH(50 mL)及冷丙酮(10 mL)洗滌，隨後冷凍乾燥，得到標題化合物(2.59 g，38%)，該化合物未作進一步純化用於下一步驟。步驟 b) 環丙基 (3- 羥基苯基 ) 甲酮 ( 苯酚 12) 在室溫下，將NaH(60%礦物油分散液)(1.13 g，28.2 mmol)分批添加至碘化三甲基氧化鋶(6.20 g，28.2 mmol)在DMSO(100 mL)中之攪拌懸浮液中。1小時後，在攪拌及冷卻下，分批添加固體Ph12-a(2.59 g，11.3 mmol)。使反應混合物在室溫下攪拌40小時，隨後傾入冷水(200 mL)中並利用DCM(3×100 mL)萃取。利用飽和NH₄ Cl水溶液(2×100 mL)洗滌有機相，乾燥(Na₂ SO₄ )，過濾並濃縮。所得粗產物藉由矽膠管柱層析(MeOH/DCM)加以純化以得到標題化合物(883 mg，48%)。苯酚 13

步驟 a) 環丙基 (4- 羥基苯基 ) 甲酮 (Ph13) 在約30分鐘內，將對羥基-γ-氯丁醯苯酮(4.95 g)分批添加至NaOH溶液(8 mL，水溶液，50% w/w)中，隨後添加NaOH(35 mL，水溶液，25% w/w)，隨後一次性添加對羥基-γ-氯丁醯苯酮(4.95 g)。使溫度降到140℃並添加NaOH(8 g)。90分鐘後，添加H₂ O(10 mL)，並再過60分鐘後，使反應混合物冷卻，利用H₂ O稀釋並利用HOAc(約27-30ml)中和到pH約7。過濾所形成之沈澱，利用H₂ O洗滌並真空乾燥。在40℃下，使固體在CHCl₃ (200 ml)中研磨10分鐘，隨後在室溫下隔夜。在30分鐘內將漿液加熱到40℃，隨後過濾。乾燥濾液(MgSO₄ )，過濾並濃縮至約70 ml。添加己烷並形成油狀物，油狀物最終變成晶體。過濾漿液，利用CHCl₃ /己烷洗滌固體並乾燥，得到標題化合物(4.15 g，51%)。苯酚 14

步驟 a) 3-(1- 羥基 -2,2- 二甲基丙基 ) 苯酚 (Ph14-a) 在30分鐘內，將t.Bu-MgBr(1.5當量)逐滴添加至3-羥基苯甲醛(2.00 g，16.4 mmol)在乙醚(20 mL)中之冷(-10℃)混合物中。添加期間，添加THF(20 mL)。使該混合物達到23℃並攪拌6小時。添加更多t.Bu-MgBr(0.7當量)並使該混合物攪拌隔夜，隨後冷卻並利用飽和NH₄ Cl水溶液淬滅該反應。將EtOAc添加至該混合物中，隨後添加1 M HCl水溶液直至得到均勻混合物。分離各相並利用鹽水洗滌有機相，乾燥(Na₂ SO₄ )，過濾並濃縮。所得粗產物藉由管柱層析加以純化以得到標題化合物(1.1 g，37%)。步驟 b) 1-(3- 羥基苯基 )-2,2- 二甲基丙 -1- 酮 (Ph14) 相繼將3 Å MS及氯鉻酸吡錠(PCC)(1.97 g，9.15 mmol)以及無水DCM(5 mL)添加至烘箱乾燥之圓底燒瓶中。使該混合物在20℃下攪拌5分鐘，隨後緩慢地添加AA8019(1.10 g，6.10 mmol)在DCM(5 mL)中之混合物。完全氧化後，經由矽藻土墊過濾混合物，利用乙醚洗滌墊子。濃縮濾液。粗產物藉由管柱層析加以純化以得到標題化合物(402 mg，37%)。MS 179.25 [M+H]⁺ 。苯酚 15

1-(4- 羥基苯基 )-2,2- 二甲基丙 -1- 酮 (Ph15) 根據苯酚14之製備所述之程序使4-羥基苯甲醛(3 g，24.6 mmol)反應，得到標題化合物(538 mg，17%)。胺基酸 1

步驟 a) (S)-(S)-2-(( 第三丁氧基羰基 ) 胺基 ) 丙酸第二丁酯 (AA1-a) 將L-Boc-丙胺酸(2.18 g，11.5 mmol)溶於無水DCM(40 mL)中並添加醇(R)-丁-2-醇(938 mg，12.6 mmol)。使該混合物冷卻至約5℃並一次性添加EDC(3.31 g，17.2 mmol)，隨後逐份添加DMAP(140 mg，1.15 mmol)。使該混合物保持在室溫下並攪拌隔夜，隨後利用乙酸乙酯(約300 ml)稀釋並利用飽和碳酸氫鈉溶液洗滌有機相三次並利用鹽水洗滌一次。經硫酸鈉乾燥有機相並在減壓下濃縮。產物藉由矽膠層析利用異己烷及10%乙酸乙酯溶離進行分離以得到標題化合物(2.78 g，98%)。步驟 b) (S)-(S)-2- 胺基丙酸第二丁酯 (AA1-b) 在65℃下，使AA1-a(2.77 g，11.3 mmol)及對甲苯磺酸單水合物(2.15 g，11.3 mmol)在EtOAc(45 mL)中之混合物攪拌16小時，隨後在減壓下濃縮。所得殘餘物自乙醚中結晶，得到標題化合物(3.20 g，89%)。胺基酸 2

(S)-(R)-2- 胺基丙酸戊 -2- 基酯 (AA2) 根據AA1之製備所述之程序，但使用(R)-戊-2-醇替代(R)-丁-2-醇，得到標題化合物(4.6 g)。胺基酸 3

(S)-(S)-2- 胺基丙酸戊 -2- 基酯 (AA3) 根據AA1之製備所述之程序，但使用(S)-戊-2-醇替代(R)-丁-2-醇，得到標題化合物(8.3 g)。製備以下中間物並可用於製備本發明化合物：中間物 1

步驟 a) (R)-2-(( 第三丁氧基羰基 ) 胺基 ) 丙酸 4- 氟苄酯 (I-1a) 將Boc-L-AlaOH(19.92 mmol)、DMAP(1.99 mmol)及(4-氟苯基)甲醇(23.9 mmol)溶於CH₂ Cl₂ (100 mL)中。相繼將三乙胺(23.9 mmol)及EDCI(23.9 mmol)添加至此溶液中並在N₂ 下，在室溫下，將所得反應混合物攪拌隔夜。利用CH₂ Cl₂ (100 mL)稀釋該反應混合物，利用飽和NaHCO₃ 水溶液(2×50 mL)，飽和NaCl水溶液(2×50 mL)洗滌，乾燥(Na₂ SO₄ )並濃縮。所得殘餘物藉由矽膠管柱層析利用正己烷-EtOAc(95:5至60:40)溶離加以純化以得到呈白色蠟樣固體狀之標題化合物(4.44 g)。MS: 296 [M-H]^- 。步驟 b) (R)-2- 胺基丙酸 4- 氟苄酯 (I-1b) 將化合物I-1a(14.93 mmol)溶於4M HCl/二噁烷(40 mL)中並在室溫下攪拌30分鐘並蒸發至乾，得到呈白色粉末狀之標題化合物之鹽酸鹽(3.4 g)。MS: 198 [M+H]⁺ 。步驟 c) (2R)-2-(( 氯 ( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸 4- 氟苄酯 (I-1) 在-78℃下，將PhOPOCl₂ (4.28 mmol)逐滴添加至化合物I-5b(4.28 mmol)之CH₂ Cl₂ 溶液中，隨後逐滴添加三乙胺(8.56 mmol)。在Ar下，在-78℃下，攪拌所得反應混合物並保持室溫下隔夜。使反應混合物在矽膠上蒸發並藉由層析(正己烷/EtOAc(88:12)-(0:100))加以純化以得到標題化合物(769 mg)。³¹ P-NMR (CDCl₃ ) δ: 7.85 (s)及7.54 (s) (R_P 及S_P 非對映異構體)。中間物 2

步驟 a) (S)-(R)-2-(( 第三丁氧基羰基 ) 胺基 ) 丙酸第二丁酯 (I-2a) 將L-Boc-丙胺酸(2.18 g，11.5 mmol)溶於無水DCM(40 mL)中並添加醇(R)-丁-2-醇(938 mg，12.6 mmol)。將該混合物冷卻至約5℃並一次性添加EDC(3.31 g，17.2 mmol)，隨後分批添加DMAP(140 mg，1.15 mmol)。使該混合物保持在室溫下並攪拌隔夜，隨後利用乙酸乙酯(約300 ml)稀釋並利用飽和碳酸氫鈉溶液洗滌有機相三次並利用鹽水洗滌一次。經硫酸鈉乾燥有機相並在減壓下濃縮。產物藉由矽膠層析利用異己烷及10%乙酸乙酯溶離進行分離以得到標題化合物(2.78 g，98%)。步驟 b) (S)-(R)-2- 胺基丙酸第二丁酯 (I-2b) 在65℃下，將I-10a(2.77 g，11.3 mmol)及對甲苯磺酸單水合物(2.15 g，11.3 mmol)在EtOAc(45 mL)中之混合物攪拌16小時，隨後在減壓下濃縮。所得殘餘物自乙醚中結晶，得到標題化合物(3.20 g，89%)。步驟 c) (2S)-(R)-2-(((4- 硝基苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸第二丁酯 (I-2) 在氮氣下，在-30℃下，將二氯磷酸苯酯(1當量)添加至化合物I-10b(3.15 g，9.92 mmol)在DCM(75 ml)中之溶液中，隨後逐滴添加三乙胺(2當量)。使該混合物保持室溫並攪拌隔夜，隨後冷卻至約5℃並以固體添加4-硝基苯酚(1當量，15 mmol)，隨後逐滴添加三乙胺(1 eq g，15 mmol)，並在室溫下攪拌該混合物4小時，隨後減壓下濃縮，利用乙酸乙酯(40 ml)及乙醚(40 ml)稀釋並保持在室溫下隔夜。過濾出三乙胺-HCl鹽並在減壓下濃縮濾液。所得殘餘物藉由矽膠管柱層析利用異己烷-乙酸乙酯溶離進行純化以得到標題化合物(4.19 g，79%)。以下化合物係根據製備I-2所述之程序利用適宜醇製備：

中間物 6 ，非對映異構體 -1 及 -2 利用SFC分離化合物I-6之兩個非對映異構體，得到I-6-非對映異構體-1及I-6-非對映異構體-2。中間物 7

步驟 a) (S)-2- 胺基丙酸環辛酯 (I-7a) 將對甲苯磺酸單水合物(3.6 g，19.1 mmol)添加至L-丙胺酸(1.7 g，19.1 mmol)及環辛醇(25 ml，191 mmol)在甲苯(100 ml)中之漿液中。使該反應混合物在迴流溫度下加熱25小時並利用Dean-Stark分離器自反應中移除水。減壓下濃縮該混合物並使殘餘物保持在真空下隔夜。將乙醚(100 ml)添加至殘餘物(27 g)中。藉由過濾收集白色沈澱，利用乙醚(3×50 ml)洗滌並在真空下乾燥以得到標題化合物(4.84 g，68%)。步驟 b) (2S)-2-(((4- 硝基苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸環辛酯 (I-7) 根據製備I-2步驟c所述之方法使化合物I-7a反應，得到標題化合物(4.7 g，76%)。中間物 8

(2S)-2-(((4- 硝基苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸環庚酯 (I-22) 按照製備化合物I-7所述之程序，但使用環庚醇(27 ml，224 mmol)替代環辛醇，得到標題化合物(5.72 g，55%)。中間物 9

按照製備I-2步驟c所述之程序，但使用(S)-2-胺基丙酸環己基酯替代(S)-2-胺基丙酸3,3-二甲基丁酯，得到標題化合物(10.6 g，82%)。中間物 10

(S)-2-(( 二 (4- 硝基苯氧基 ) 磷醯基 ) 胺基 ) 丙酸 2- 乙基丁酯 (I-10) 將(S)-2-胺基丙酸2-乙基丁酯(5 g，14.49 mmol)添加至氯磷酸雙(4-硝基苯基)酯(6.14 g，17.1 mmol)在DCM(50 ml)中之溶液中，在冰浴中使該混合物冷卻並逐滴添加Et₃ N(4.77 mL，34.2 mmol)。15分鐘後，移除冷卻並在23℃下攪拌該反應混合物，直至根據TLC顯示反應完全。隨後添加乙醚，過濾混合物並濃縮濾液且藉由矽膠管柱層析進行純化以得到標題化合物(2.05 g，82%)。中間物 11

步驟 a) (S)-2- 胺基丙酸異丙酯 (I-11a) 在0℃下，將SOCl₂ (29 mL，400 mmol)逐滴添加至L-丙胺酸之HCl鹽(17.8 g，200 mmol)在異丙醇(700 mL)中之懸浮液中。使該懸浮液在室溫下攪拌隔夜，隨後濃縮，得到標題化合物(29.2 g，87%)。步驟 b) (2S)-2-(((((S)-1- 異丙氧基 -1- 側氧基丙 -2- 基 ) 胺基 )(4- 硝基苯氧基 ) 磷醯基 )- 胺基 ) 丙酸異丙酯 (I-11) 在-60℃下，將二氯磷酸4-硝基苯酯(1.8 g，7 mmol)在DCM中之溶液逐滴添加至胺I-11a (2.35 g，14 mmol)及三乙胺(7.7 mL，56 mmol)在DCM中之溶液中。使該反應混合物保持室溫，攪拌隔夜，濃縮隨後利用乙酸乙酯及乙醚稀釋並保持在室溫下隔夜。過濾出三乙胺HCl鹽，減壓下濃縮濾液且所得殘餘物藉由矽膠層析利用異己烷-乙酸乙酯溶離進行純化，得到標題化合物(1.6 g，50%)。中間物 12

步驟 a) (S)-2-(( 第三丁氧基羰基 ) 胺基 ) 丙酸新戊酯 (I-12a) 在-5℃下，將EDAC及DMAP分批添加至Boc-丙胺酸(18.9 g，100 mmol)及新戊醇(13.0 mL，120 mmol)在DCM(200 mL)中之溶液中。使該反應混合物保持在室溫下並攪拌72小時。添加EtOAc(700 mL)並利用飽和NaHCO₃ 溶液洗滌有機相三次且利用鹽水洗滌一次，隨後濃縮。所得殘餘物藉由管柱層析利用己烷-EtOAc 90/10至80/20溶離進行純化以得到標題化合物(21 g，81%)。步驟 b) (S)-2- 胺基丙酸新戊酯 (I-12b) 在-65℃下，將對甲苯磺酸(15.6 g，82.0 mmol)添加至Boc保護之胺I-12a(21.1 g，82.0 mmol)在EtOAc(330 mL)中之溶液中。使該反應混合物在-65℃下攪拌8小時，隨後保持室溫隔夜。隨後，過濾混合物並濃縮以得到標題化合物(21 g，78%)。(2S)-2-(((((S)-1-( 新戊基氧基 )-1- 側氧基丙 -2- 基 ) 胺基 )(4- 硝基苯氧基 )- 磷醯基 ) 胺基 ) 丙酸新戊酯 (I-12) 在-50℃下，在1小時內，將4-硝基苯酚二氯磷酸酯逐滴添加至胺I-12b(3.90 g，24.5 mmol)在DCM(100 mL)中之溶液中。使該反應混合物保持室溫，攪拌隔夜，濃縮隨後利用乙醚稀釋並於室溫下隔夜。過濾該混合物，減壓下濃縮濾液且所得殘餘物藉由矽膠層析於利用異己烷-乙酸乙酯溶離進行純化以得到標題化合物(4.8 g，77%)。中間物 32

(2S)-(R)-2-((( 全氟苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸第二丁酯 (I-32) 在-70℃下，在氮氣下，在15分鐘內將Et₃ N(10.9 mL，78.1 mmol)逐滴添加至(S)-(R)-2-胺基丙酸第二丁酯之pTs鹽(12.0 g，37.7 mmol)在DCM(50 mL)中之攪拌溶液中。在1小時內，將二氯磷酸苯酯(5.61 mL，37.7 mmol)在DCM(50 mL)中之溶液添加至該混合物中。在-70℃下，使反應混合物再攪拌30分鐘，隨後在2小時內升溫至0℃並攪拌1小時。在20分鐘內，將五氟苯酚(6.94 g，37.7 mmol)及Et₃ N(5.73 mL，41.1 mmol)在DCM(30 mL)中之溶液添加至該混合物中。使粗製混合物在0℃下攪拌18小時，隨後濃縮。將殘留物溶於THF(100 mL)中，過濾出不溶物並利用THF洗滌多次。蒸發溶劑並來利用第三丁基甲醚研磨殘留物。過濾出不溶物並利用第三丁基甲醚洗滌。濃縮合併之濾液並利用正己烷/EtOAc(80:20；100 mL)超音處理粗製固體。過濾固體，利用正己烷/ EtOAc(80:20)洗滌，得到呈白色固體狀之標題化合物之純磷立體異構體(2.3 g，13%)。中間物 33

(2S)-2-((( 全氟苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸乙酯 (I-33) 標題化合物之純磷立體異構體係根據針對I-32所述之方法製備，但以(S)-2-胺基丙酸乙酯之HCl鹽(11.0 g，71.1 mmol)為起始物。產率為8.56 g，27%。中間物 34

(2S)-2-((( 全氟苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸 2- 乙基丁酯 (I-34) 標題化合物之純磷立體異構體係根據針對I-32所述之方法製備，但以(S)-2-胺基丙酸2-乙基丁酯之pTs鹽(18.8 g，54.4 mmol)為起始物。產率為27.0 g，99%。 LC-MS 496.44 [M+H]⁺ 。中間物 35

(2S)-2-((( 全氟苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸丁酯 (I-35) 將二氯磷酸苯酯(12.4 mL，83.1 mmol)添加至(S)-2-胺基丙酸丁酯(26.4 g，83.1 mmol)在二氯甲烷(200 mL)中之冷(-20℃)漿液中。使該混合物攪拌10分鐘，隨後在15分鐘內逐滴添加Et₃ N(25.5 mL，183 mmol)。在-20℃下攪拌該混合物1小時，隨後在0℃下攪拌30分鐘。使該混合物在冰浴中冷卻並添加全氟苯酚(15.3 g，0.08 mol)，隨後逐滴添加Et₃ N(11.6 mL，0.08 mol)。攪拌該混合物隔夜並緩慢升至20℃。添加乙醚並藉由矽藻土過濾該混合物，濃縮並藉由矽膠管柱層析利用石油醚/EtOAc(9:1 -＞ 8:2)溶離進行純化。合併適宜溶離份，濃縮並自石油醚/EtOAc(9:1)中結晶，得到呈白色固體狀之標題化合物之純磷立體異構體(2.23 g，5.8%)。中間物 36

步驟 a) L- 丙胺酸異丙酯鹽酸鹽 (I-36a) 在-7至0℃下，經30分鐘之時段，在冷卻下，將亞硫醯氯(80.2 g，0.674 mol，1.5當量)添加至2-丙醇(400 mL)中，隨後在0℃下，添加L-丙胺酸(40.0 g，0.449 mol)。將流量指示器及具有27.65%氫氧化鈉(228 g)與水(225 g)之混合物之洗滌器連接到出口。使該反應混合物在67℃下攪拌2小時，隨後在70℃下攪拌1小時並在20-25℃下攪拌隔夜。在47-50℃下，減壓(250 - 50毫巴)下自60℃浴中蒸餾出反應混合物。當蒸餾變得非常緩慢時，將甲苯(100 mL)添加至殘留油狀物中，繼續在48-51℃下，在減壓(150 - 50毫巴)下自60℃浴中蒸餾，直至變得非常緩慢。將第三丁基甲醚(tBME)(400 mL)添加至殘留油狀物中，在34-35℃下，在有效攪拌下，對兩相系統種晶。當觀察到結晶時，經一小時之時段將該混合物冷卻至23℃，藉由過濾分離出沈澱。利用tBME(100 mL)洗滌濾餅並在減壓下不加熱下乾燥到恆重，得到呈白色固體狀之標題化合物(67.7 g，90%)。步驟 b) (S)-2-(((S)-( 全氟苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸異丙酯 (I-36) 在0℃下，在氮氣下，將二氯磷酸苯酯(62.88 g，0.298 mol，1.0當量)添加至L-丙胺酸異丙酯鹽酸鹽(50.0 g，0.298 mol)在DCM(310 mL)中之溶液中，該添加藉由用DCM(39 mL)洗滌完成。使該混合物冷卻並經70分鐘之時段，在冷卻下，添加三乙胺(63.35 g，0.626 mol，2.1當量)，保持溫度不超出-14℃，該添加藉由用DCM(39 mL)洗滌完成。在-15至-20℃下，將該混合物攪拌1小時，隨後加熱至-8℃並經42分鐘之時段，添加全氟苯酚(60.38 g，0.328 mol，1.1當量)與三乙胺(33.19 g，0.328 mol，1.1當量)在DCM(78 mL)中之溶液，保持溫度不超出0℃，該添加藉由用DCM(39 mL)洗滌完成。使該混合物在0℃下攪拌1小時，隨後在+5℃下攪拌隔夜。藉由過濾移除所形成之沈澱，並利用DCM(95 mL)洗滌濾餅。在5℃下，利用水(2×190 mL)洗滌合併之濾液。在32-38℃下，減壓(650 - 600毫巴)下蒸餾有機相，繼續蒸餾直至剩餘體積為約170 mL。得到部分結晶物。添加乙酸乙酯(385 mL)，使所得之澄清溶液在43-45℃下在減壓(300 - 250毫巴)下蒸餾。繼續蒸餾直至剩餘體積為約345 mL。將澄清溶液冷卻至36℃，藉由添加根據J. Org. Chem., 2011, 76, 8311 – 8319所述製備之(S)-2-(((S)-(全氟苯氧基)(苯氧基)磷醯基)胺基)丙酸異丙酯之晶種(20 mg)誘導結晶。經1小時之時段，將該混合物冷卻至27℃，隨後經47之時段添加正庚烷(770 mL)，再攪拌該混合物37分鐘。添加三乙胺(6.03 g，0.2當量)，使該混合物在23-25℃下攪拌隔夜。藉由過濾分離沈澱。利用乙酸乙酯：正庚烷(1:9，80 mL)洗滌濾餅並在減壓(低於0.1毫巴)下不加熱下乾燥至恆重，得到呈白色結晶物狀之標題化合物(75.64 g，56%)。¹ H NMR (CDCl₃ , 300 MHz) δ 7.38-7.32 (m, 2 H), 7.27-7.24 (m, 2 H), 7.23-7.19 (m, 1 H), 5.10-4.98 (m, 1 H), 4.20-4.08 (m, 1 H), 4.03-3.96 (m, 1 H), 1.46 (dd, 7.2, 0.6 Hz, 3 H), 1.26-1.23 (2xd, 6 H)；¹³ CNMR (CDCl₃ , 100 MHz) δ 172.7 (d, J = 8.8 Hz), 150.4 (d, J = 7.1 Hz), 143.4-143.0 (m), 141.0-140.2 (m), 140.0-139.8 (m), 137.6-137.2 (m), 136.8-136.2 (m), 130.0 (d, J = 0.82 Hz), 125.8 (d, J = 1.4 Hz), 120.3 (d, J = 5.0 Hz), 69.8, 50.6, (d, J = 1.9 Hz), 21.8 (d, J = 1.9 Hz), 21.2 (d, J = 4.4 Hz)；標題化合物之結晶性質及NMR光譜資料與公佈資料(J. Org. Chem., 2011, 76, 8311-8319)一致，因此證實標題化合物之磷原子之S立體化學。中間物 37

步驟 a) (S)-2- 胺基丙酸環己酯 (I-37a) 將乙醯氯(4.2 mL，59.3 mmol)逐滴添加至環己醇之攪拌溶液(50 ml)中，隨後添加L-苯丙胺酸(4.0 g，24.2 mmol)。將該反應混合物加熱至100℃，保持16小時，隨後減壓下濃縮，利用乙醚/己烷(1:1)研磨並乾燥以得到呈白色固體狀之標題化合物(6 g，88%)，該化合物未作進一步純化用於下一步驟。步驟 b) (S)-2-(((S)-( 全氟苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸環己酯 (I-37) 在-70℃下，經30分鐘，將三乙胺(7.17 mL，51.5 mmol)逐滴添加至化合物I-37a(7.0 g，24.6 mmol)在無水DCM(42 mL)中之攪拌溶液中，隨後經1小時添加二氯磷酸苯酯(5.15 g，34.5 mmol)在無水DCM(21 mL)中之溶液。在-70℃下，使該反應混合物再攪拌30分鐘，隨後經2小時升溫至0℃並攪拌1小時。經1小時，將全氟苯酚(4.94 g，26.8 mmol)及三乙胺(3.74 mL，26.8 mmol)在無水DCM(28 mL)中之溶液添加至該混合物中。使該混合物在0℃下攪拌4小時，隨後在5℃下攪拌16小時。過濾反應混合物並在減壓下濃縮濾液。將殘餘固體溶於EtOAc(300 mL)中，利用水(50 mL)洗滌，乾燥並在減壓下移除溶劑。利用20% EtOAc之己烷溶液研磨所得固體，過濾，利用己烷洗滌並乾燥以得到呈固體狀之單一非對映異構體之標題化合物(3.0 g，21%)。中間物 38

(2S)-2-(((4- 硝基苯氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸異丙酯 (I-38) 在-78℃下，經30分鐘之時段，將苯酚(1.86 g，19.8 mmol)及三乙胺(3 mL，21.8 mmol)在無水DCM(50 mL)中之溶液添加至二氯磷酸4-硝基苯酯(5 g，19.8 mmol)在無水DCM(40 ml)中之攪拌溶液中。使該混合物在此溫度下攪拌60分鐘，隨後在-5℃下，經15分鐘之時段，轉移到包含化合物(S)-2-胺基丙酸異丙酯(3.3 g，19.8 mmol)在無水DCM(40 mL)中之溶液之另一燒瓶中。在-5℃下，經20分鐘之時段，將第二份TEA(6 mL，43.3 mmol)添加至該混合物中。使該混合物在0℃下攪拌3小時，隨後減壓下移除溶劑。將殘餘物溶於EtOAc(200 mL)中並利用水(50 mL)洗滌，經Na₂ SO₄ 乾燥並在減壓下移除溶劑以得到呈油狀之粗產物，隨後藉由管柱層析利用0-20% EtOAc/己烷梯度以及230-400目矽膠進行純化以得到約1:1比率之非對映異構體之混合物。藉由SFC分離非對映異構體，得到標題化合物，呈固體狀之異構體1(1.5 g，20%)及異構體2(1.5 g，18%)。表1中所列之化合物利用適宜之胺基酸酯及苯酚，根據製備中間物1-38所述之程序製備，並分離非對映異構體。表1

實例 1

步驟 a) 乙酸 ((2S,4S)-4-(2,4- 二側氧基 -3,4- 二氫嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲酯 (1a) 在125℃下，使化合物Tr-8(0.15 g，0.41 mmol)、1,2-二甲氧基乙烷(1.5 mL)及水(0.96 mL)之混合物在密封管中加熱48小時。反應完全後(TLC)，將該反應混合物冷卻至室溫並在減壓下移除溶劑。粗製殘餘物藉由管柱層析在230-400矽膠上利用3-7% MeOH/DCM梯度進行純化以得到呈固體狀之化合物1a(0.08 g，80%)以及呈固體狀之化合物1b(0.02 g)。步驟 b) 1-((2S,4S)-2-( 羥甲基 )-1,3- 二氧雜環戊烷 -4- 基 ) 嘧啶 -2,4(1H,3H)- 二酮 (1b) 在室溫下，使含化合物1a(0.08 g，0.31 mmol)之飽和NH₃ 之MeOH溶液(1.6 mL)在密封管中攪拌4小時。反應完全後(TLC)，減壓下移除溶劑且殘餘物藉由管柱層析在60-120矽膠上利用5-7% MeOH/DCM進行純化以得到呈固體狀之標題化合物(0.06 g，90%)。步驟 c) (2S)-2-(((((2S,4S)-4-(2,4- 二側氧基 -3,4- 二氫嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸異丙酯 (1c) 在-5℃下，將氯化第三丁基鎂(0.57 mL，0.98 mmol，1.7 M THF)逐滴添加至化合物1b(60 mg，0.28 mmol)在DMPU(0.6 mL)中之攪拌溶液中。使該混合物在-5℃下攪拌30分鐘，隨後在室溫下攪拌30分鐘。在-5℃下，添加((全氟苯氧基)(苯氧基)磷醯基)-L-丙胺酸異丙酯(0.25 g，0.56 mmol)在無水THF(2.5 mL)中之溶液並在室溫下攪拌該反應混合物8小時。反應完全後(TLC)，加水(15 mL)並利用EtOAc(30 mL)萃取該混合物。利用飽和氯化鈉溶液(10 mL)洗滌有機相，乾燥(Na₂ SO₄ )，過濾並濃縮，所得粗製物藉由管柱層析在230-400矽膠上利用4-5% MeOH/DCM梯度進行純化以得到呈固體狀之標題化合物(55 mg，38%)。MS (ES+) [484.0]⁺ 。¹ H NMR (DMSO-d₆ , 400 MHz) δ 1.15-1.20 (10H), 3.73-3.75 (1 H), 4.11-4.27 (4H),4.84-4.90 (1H), 5.14 (1H), 5.51-5.53 (1H), 6.06-6.12 (1H), 6.26-6.27 (1H), 7.17-7.23 (3H), 7.36-7.40 (2H), 7.57-7.60 (1H), 11.37 (1H)。實例 2

(2S)-2-(((((2S,4S)-4-(4- 胺基 -2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸異丙酯 (2) 根據實例1步驟c所述之程序，使曲沙他濱(TR-9)(50 mg，0.23 mmol)與磷酸化劑I-36(0.26 g，0.58 mmol)反應，得到呈固體狀之標題化合物(30 mg，26%)。MS (ES+) 483.34 [M+H]⁺ 。¹ H NMR (DMSO-d₆ , 400 MHz) δ 1.14-1.24 (9H), 3.32-3.38 (1H), 4.05-4.21 (4H), 4.84-4.26 (1H), 5.14 (1H), 5.68-5.70 (1H), 6.07-6.13 (1H), 6.23-6.25 (1H), 7.16-7.24 (5H), 7.34-7.39 (2H), 7.59-7.61 (1H)。實例 3

(2S)-2-(((((2S,4S)-4-(4- 胺基 -2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸異丙酯 (3) 根據實例1步驟c所述之程序，使曲沙他濱(50 mg，0.23 mmol)與磷酸化劑I-38(0.24 g，0.58 mmol)反應，得到呈固體狀之標題化合物(40 mg，35%)。MS (ES+) 481.0 [M-H]^- 。¹ H NMR (DMSO-d₆ , 400 MHz) δ 1.14-1.20 (9H), 3.76-3.77 (1H), 4.10-4.18 (2H), 4.22-4.25 (2H), 4.84-4.87 (1H), 5.17-5.186 (1H), 5.69-5.70 (1H), 6.03-6.08 (1H), 6.24-6.26 (1H), 7.17-7.25 (5H), 7.36-7.40 (2H), 7.62-7.64 (1H)。實例 4

(2S)-2-(((((2S,4S)-4-(4- 胺基 -2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸異丙酯 (4) 根據實例1步驟c所述之程序，使曲沙他濱(50 mg，0.23 mmol)與磷酸化劑I-37(0.33 g，0.58 mmol)反應，得到呈固體狀之標題化合物(30 mg，22%)。MS (APCI) 599.47 [M+H]⁺ 。表2中所列之化合物利用適宜中間物，根據實例1步驟c所述之程序以純非對映異構體製備，I-編號非對映異構體-1或I-編號非對映異構體-2。表2

類似地，表3中所列之化合物利用適宜中間物，根據實例1步驟c所述之程序以純非對映異構體製備。表3

記錄所有示例化合物之NMR及MS數據從而確證其結構。實例 35

(2S)-2-(((((2S,4S)-4-(2- 側氧基 -4- 棕櫚醯胺嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸異丙酯 (35 非對映異構體 1 及 35 非對映異構體 -2) 根據WO2008/030373中所述方法，利用棕櫚酸酐分別醯化化合物2及3，得到標題化合物。實例 36

(2S)-2-(((((2S,4S)-4-(2- 側氧基 -4- 棕櫚醯胺嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲氧基 )( 苯氧基 ) 磷醯基 ) 胺基 ) 丙酸甲酯 (36) 根據WO2008/030373中所述之方法，利用棕櫚酸酐醯化化合物27非對映異構體-2，得到標題化合物。比較實例

步驟 a) (2S)-2-(( 雙 (4- 甲氧基苯基 )( 苯基 ) 甲基 ) 胺基 )-N-(2- 氧離子基 -1,3,2- 氧硫磷雜環戊烷 -2- 基 ) 丙醯胺 在氮氣下，將2-氯-1,3,2-氧硫磷雜環戊烷溶液(0.542 g，3.80 mmol)逐滴添加至(S)-2-((雙(4-甲氧基苯基)(苯基)甲基)胺基)丙醯胺(1.40 g，3.58 mmol)及三乙胺(0.60 ml，4.30 mol)在二氯甲烷(8 ml)中之冰冷溶液中。使反應維持在室溫並攪拌過週末。將該溶液冷卻至0℃並緩慢地添加(第三丁基過氧基)三甲基矽烷(1.16 g，7.17 mmol)之庚烷溶液。使該反應混合物攪拌90分鐘，隨後真空濃縮。使殘餘物懸浮在乙酸乙酯(10 mL)中，藉由過濾移除鹽酸鹽並真空移除溶劑。使殘餘物溶解在無水乙腈(10 mL)中且所得溶液未作進一步純化用於下一步。基於³¹ P-NMR確定定量產率及80%純度。步驟 b) ((S)-2-(( 雙 (4- 甲氧基苯基 )( 苯基 ) 甲基 ) 胺基 ) 丙醯基 ) 胺基磷酸氫 ((2S,4S)-4-(4- 胺基 -2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲酯在氮氣下，將DMAP(229 mg，1.88 mmol)添加至化合物Tr-9(100 mg，0.469 mmol)在無水嘧啶(5 mL)中之溶液中，隨後緩慢地添加(2S)-2-((雙(4-甲氧基苯基)(苯基)甲基)胺基)-N-(2-側氧基-1,3,2-氧硫磷雜環戊烷基)丙醯胺(361 mg，0.563 mmol)在無水乙腈(2 mL)中之溶液中。在氮氣下，使所得溶液在室溫下攪拌46小時，隨後濃縮。殘餘物藉由製備型HPLC在Gemini-NX 5m C18 (100×30 mm)上利用在17分鐘內20% B至80% B之梯度以35 mL/min之流速進行純化。溶劑A：95%水，5%乙腈(10 mM乙酸銨)；溶劑B：10%水，90%乙腈(10 mM乙酸銨)。合併含有產物之溶離份並冷凍乾燥，得到標題化合物(80 mg，26%)。MS (ES+) 664.26 [M+H]⁺ 。步驟 c) ((S)-2- 胺基丙醯基 ) 胺基磷酸氫 ((2S,4S)-4-(4- 胺基 -2- 側氧基嘧啶 -1(2H)- 基 )-1,3- 二氧雜環戊烷 -2- 基 ) 甲酯將水(50 mL)添加至來自先前步驟之化合物(80.5 mg，0.121 mmol)之二氯甲烷溶液中，隨後添加乙酸(500 mL)。室溫下，將該溶液攪拌12分鐘，隨後添加TFA(75 mL)並使所得溶液在室溫下攪拌5分鐘，利用甲苯(10 mL)稀釋，濃縮至乾並真空下乾燥。將殘留物溶於含10%乙腈之水(10 mL)中並利用含10%己烷之第三丁基甲醚(2×10 mL)洗滌。收集水層並冷凍乾燥隔夜以得到呈雙-TFA鹽形式之所需產物(80 mg)，該產物根據LC-MS具有約75%之純度。所需殘留物藉由製備型HPLC在Hypercarb(21.2× 100 mm, I=271 nm)上利用0%至35%乙腈/水梯度液進行進一步純化。合併含有產物之溶離份並冷凍乾燥。MS (ES+) 364.10 [M+H]⁺ 。利用¹ H及¹³ C NMR證實結構。所選擇之示例性化合物之NMR資料：化合物 8 非對映異構體 -1 ¹ H NMR (DMSO-d₆ , 400 MHz) δ 0.81-0.84 (6H), 1.20-1.22 (11H), 1.59 (1H), 3.82-3.97(3H), 4.08-4.16 (2H), 4.22-4.23 (2H), 5.16 (1H), 5.67-5.69 (1H), 6.05-6.10 (1H), 6.23-6.24 (1H), 7.16-7.23 (m, 5H), 7.34-7.38 (m, 2H), 7.60-7.62 (m, 1H)。化合物 8 非對映異構體 -2 ¹ H NMR (DMSO-d₆ , 400 MHz) δ0.81-0.84 (6H), 1.22-1.27 (11H), 1.57(1H), 3.81-3.89 (2H), 3.95-3.98 (1H), 4.05-4.07 (1H), 4.10-4.20 (3H), 5.128 (1H), 5.68-5.69 (1H), 6.13-6.14 (1H), 6.22-6.24 (1H), 7.16-7.21 (5H), 7.34-7.38 (2H), 7.58-7.60 (1H)。化合物 9 非對映異構體 -1 ³¹ P NMR (DMSO-d₆ ) δ 4.354。¹ H NMR (DMSO-d₆ , 400 MHz) δ1.24-1.26 (3H), 3.98-4.01 (1H), 4.12-4.14 (2H), 4.27-4.29 (2H), 5.00-5.08 (2H), 5.16-5.18 (1H), 5.64-5.66 (2H), 6.25-6.27 (1H), 6.34 (1H), 7.17-7.22 (2H), 7.31-7.33 (5H), 7.45-7.46 (2H), 7.55-7.59 (2H), 7.63-7.64 (1H), 7.74-7.77 (1H), 7.95-7.97 (1H), 8.08-8.11 (1H)。化合物 9 非對映異構體 -2 ³¹ P NMR (DMSO-d₆ ) δ 4.159。¹ H NMR (DMSO-d₆ , 400 MHz) δ1.25-1.26 (3H), 3.97-4.01 (1H), 4.08-4.16 (2H), 4.23-4.29 (2H), 5.04-5.16 (3H),5.65-5.66 (1H), 6.26 (1H), 6.36-6.42 (1H), 7.17-7.24 (2H), 7.326 (5H), 7.41-7.49 (2H),7.57-7.64 (3H),7.74-7.76 (1H), 7.95-7.97 (1H), 8.10-8.12 (1H)。化合物 11- 非對映異構體 -1 ¹ H NMR (DMSO-d₆ , 400 MHz) δ0.23 (9H), 0.78-0.82 (3H), 1.08-1.12 (3H), 1.20-1.22 (3H), 1.44-1.49 (2H), 3.77-3.79 (1H), 4.09-4.23 (4H), 4.67-4.72 (1H), 5.16-5.16 (1H), 5.69-5.70 (1H), 6.04-6.10 (1H), 6.23-6.25 (1H), 7.15-7.24 (4H), 7.48-7.50 (2H), 7.61-7.63 (1H)。化合物 11 非對映異構體 -2 ¹ H NMR (DMSO-d₆ , 400 MHz) δ0.22-0.24 (9H), 0.78-0.82 (3H), 1.10-1.11 (3H), 1.22-1.24 (3H), 1.46-1.50 (2H), 4.05-4.07 (1H), 4.11-4.22 (4H), 4.70-4.71 (1H), 5.14 (1H), 5.69-5.71 (1H), 6.07-6.11 (1H), 6.23-6.25 (1H), 7.16-7.24 (4H), 7.49-7.51 (2H), 7.60-7.62 (1H)。為了前藥靶向肝，前藥之正確處理係至關重要的。前藥應在腸液中穩定，並在肝臟中在首過代謝中經肝酶處理以形成單磷酸鹽。隨後，所形成之單磷酸鹽在肝細胞中經細胞激酶合成代謝為活性三磷酸鹽物質。此外，抗癌藥物應對增殖細胞應有毒性。評價化合物之此等性質之適宜方法為例如如下所述。在人類腸 S9 部分 (HIS9) 中以及在人類肝 S9 部分 (HLS9) 中之穩定性 在DMSO中製備各測試化合物之儲備溶液(10 mM)並儲存在-20℃下。開始實驗前，使測試化合物在50%乙腈水溶液中稀釋至500 µM。在50 mM磷酸鉀緩衝液(pH 7.4)中以250 μL總體積製備含有5 mM MgCl₂ 、1 mM NADPH及5 μM測試化合物之反應混合物。藉由添加人類肝或腸S9部分使得最終濃度為0.4 mg蛋白/mL起始反應(Xeno Tech)。使反應混合物在定軌振盪器上在37℃下培養。在所需時間點(0、10、30及60分鐘)，取50 μL等分試樣並藉由與150 μL含內標之乙腈混合停止反應。由500 µM溶液藉由將溶液在煮沸之人類S9(0.4 mg蛋白/mL)、5 mM MgCl₂ 及50 mM磷酸鉀緩衝液(pH 7.4)中稀釋至5 µM之最終濃度來製備各測試化合物之標準溶液。將標準及樣品置於冰上30分鐘隨後以3 000 g在10℃下離心20分鐘，隨後將10 µL上清液與200 µL 50%乙腈水溶液混合。將含0.5 µM各測試化合物之50%乙腈水溶液注入LC/MS-MS中以測定子離子、去簇電壓(DP)、碰撞能量(CE)及碰撞室出口電壓(CXP)以進行LC/MS-MS法。利用具有QTRAP5500系統之C18柱分離化合物。流動相係由溶劑A(98%水、2%乙腈、0.1%乙酸或10 mM乙酸銨)及溶劑B(80%乙腈、20%水、0.1%乙酸或10 mM乙酸銨)組成。藉由0%至100%之溶劑B梯度進行化合物之溶離。注入5 µL標準點及樣品以進行QTRAP5500分析。基於各時間點之峰面積相比於設定為5 µM之標準測定母化合物之量。利用Excel軟體，由測試化合物之消失曲線測定固有清除率(CL_int )及半衰期(t_1/2 )。細胞毒性分析 在添加化合物前24小時，接種細胞。將各測試化合物(自100 μM連續稀釋)添加至Huh7(1.5×10⁴ 個細胞/孔)或 HepG2(1.5×10⁴ 個細胞/孔)中，並在37℃下培養5天。僅培養基對照用於測定最小吸收值以及未處理細胞值。在生長期結束時，將來自Polysciences Europe GmbH之XTT染料添加至各孔中。在600 nM之參考波長下利用Sunrise(Tecan)讀取450 nm下之吸收值，將僅培養基對照用作空白。藉由比較對化合物濃度繪製之抑制程度(相對於細胞對照)確定50%抑制值(CC₅₀ )。將稀釋系列之結果擬合成s型劑量-反應曲線。在此等分析中評價本發明化合物以評價在人類腸S9部分(HIS9)及人類肝S9部分(HLS9)中之穩定性，以及在HUH7、HEP3B及HEPG2細胞中之細胞毒性。結果彙總在表B1中。表B1

na =不可用三磷酸鹽形式分析 分析中，以一式三份測試各化合物。在12孔板中使用新鮮人類接種之幹細胞(Biopredic, France)。使各孔接種0.76×10⁶ 個細胞並與10 µM化合物(0.1% DMSO)之DMSO溶液在1 mL培養基中在CO₂ 培養箱中在37℃下培養8小時。將在具有抗生素及10%胎牛血清之DMEM中生長之Huh7細胞接種於12孔板中，2×10⁵ 個細胞/孔。24小時後，添加1 mL含10 µM化合物之培養基並再培養細胞6-8小時。藉由利用1 mL冰冷漢克平衡溶液pH 7.2洗滌各孔兩次，隨後添加0.5 mL冰冷70%甲醇使培養停止。添加甲醇後，立即藉由細胞刮刀將細胞層自孔底脫離並利用自動吸管來回吸取5-6次。將細胞懸浮液轉移到玻璃瓶並在-20℃下儲存隔夜。隨後，使樣品渦旋振盪並在10℃下以14000 rpm在Eppendorf離心機5417R中離心10分鐘，各樣品係由多種濃度之前藥、游離核苷、以及單-、二-及三磷酸鹽組成。藉由插片，將上清液轉移到2 mL玻璃小瓶中並根據以下進行生物分析：將內標(印地那韋(Indinavir))添加至各樣品中並在耦合至QTRAP 5000質譜儀之兩個柱系統上分析樣品(10 µL注射體積)。兩個柱系統係由兩個二元泵，X及Y，兩個轉換閥及自動進樣器組成。所用的兩個HPLC柱為Synergy POLAR-RP 50*4.6 mm，4 µm顆粒及BioBasic AX 50*2.1 mm，5 µm顆粒。LC流速為0.4-0.6 mL/min(在再調節步驟中使用更高流速)。用於POLAR-RP柱之HPLC流動相係由10 mmol/L乙酸銨之2%乙腈溶液(流動相A)及10 mmol/L乙酸銨之90%乙腈溶液(流動相B)組成，BioBasic AX柱之HPLC流動相係由10 mmol/L乙酸銨之2%乙腈溶液(流動相C)及1%氫氧化銨之2%乙腈溶液(流動相D)組成。泵Y之HPLC梯度以0%流動相B開始並保持2分鐘。負載階段期間，使流動相通過POLAR-RP及BioBasic AX柱，並在POLAR-RP柱上捕集前藥、核苷及內標；而核苷酸(單-、二-及三磷酸)溶離到BioBasic AX柱上並捕集在此。在下一步驟中，將來自POLAR-RP柱之流切換到MS並將流動相C自泵X切換到BioBasic AX柱。利用在約兩分鐘內0% B至100% B梯度溶離POLAR-RP柱上之化合物並利用多反應監測模式(MRM)以正或負模式分析。在最後一步中，將來自BioBasic AX柱之流切換到MS並利用約7分鐘直至50% D梯度溶離磷酸鹽，並利用MRM以正或負模式分析。在最後一步中，再新調節兩個柱子。隨後，藉由與標準曲線比較確定各化合物之三磷酸鹽濃度，該等標準曲線係藉由分析具有已知濃度之三磷酸鹽之標準樣品製得。使標準在與測試樣品相同之基質上跑樣。因為肝細胞供體之間磷酸化水準之變化，所以各輪分析中需要內標化合物以將不同輪次之結果彼此排序。在說明書及以下申請專利範圍中，除非上下文另外需要，否則單詞『包含(comprise)』以及其變化形式『包含(comprises)』及『包含(comprising)』應理解為暗指包括所示整體、步驟、整體組或步驟組但不排除任何其他整體、步驟、整體組或步驟組。文中所述之所有文獻(包括專利及專利申請案)之全文係以引用之方式併入。Therefore, various embodiments and intermediates of the present invention will now be illustrated by the following examples. These examples are only used to further illustrate the present invention and do not limit the scope of the present invention in any way. The compound name was generated by ChemDraw Ultra software, Cambridgesoft, version 12.0.2. In addition to the above definitions, the following abbreviations are used in the above synthetic schemes and the following examples. If the abbreviations used in the text are not defined, they have their accepted meanings.

Preparation of Qusatabin

step 1) ((2,2- Dimethoxyethoxy ) methyl ) benzene (Tr-1) At 0°C, add benzyl bromide (56.03 mL, 0.471 mol) and NaOH (20.7 g, 0.518 mol) to 2,2-dimethoxyethanol (50 g, 0.471 mol) in DMF (200 mL) The reaction mixture was stirred and the reaction mixture was stirred at room temperature for 16 hours. After the reaction was complete (TLC), saturated sodium chloride solution (500 mL) was added and the reaction mixture was extracted with DCM (1 L), and the organic phase was dried (Na₂ SO₄ ) And concentrated and the resulting crude product was purified by silica gel column chromatography on 60-120 silica using 4-6% EtOAc/hexane to obtain the title compound (60 g, 60%) as a liquid.step 2) (5S)-5-((4S)-2-(( Benzyloxy ) methyl )-1,3- Dioxolane -4- base )-3,4- Dihydroxyfuran -2(5H)- ketone (Tr-2) L-ascorbic acid (44.9 g, 0.255 mol) was added to a solution of compound Tr-1 (60 g, 0.306 mol) in anhydrous acetonitrile (898 mL), followed by pTSA monohydrate (15.5 g, 0.076 mol) and The reaction mixture was heated at 90°C for 1 hour. After the reaction is complete (TLC), half the volume of acetonitrile is distilled off and the process is repeated twice. The solvent was completely removed and the title compound (91 g) was obtained as a mixture of stereoisomers. The product was used directly in the next step without further purification.step 3) (2R)-2-((4S)-2-(( Benzyloxy ) methyl )-1,3- Dioxolane -4- base )-2- Glycolic acid (Tr-3) At room temperature, compound Tr-2 (91.7 g, 0.297 mol) was added to K₂ CO₃ (86.3 g, 0.625 mol) in H₂ O (509 mL) in the stirred solution. Add H slowly₂ O₂ (80 mL, 0.71 mol, 30% v/v) and the solution was cooled to 0 °C and then stirred for 24 hours. The solvent was removed under reduced pressure, EtOH (100 mL) was added and the mixture was heated at reflux for 30 minutes, then filtered. EtOH (100 mL) was added to obtain a solid residue and the mixture was heated at reflux for 30 minutes (twice). The collected filtrate was concentrated under vacuum to obtain the title compound (90 g) as a solid.step 4) (2S,4S)-2-(( Phenoxy ) methyl )-1,3- Dioxolane -4- Formic acid (Tr-4a) and (2R,4S)-2-(( Phenoxy ) methyl )-1,3- Dioxolane -4- Formic acid (Tr-4b) Within 30 minutes, sodium hypochlorite (650 ml, 0.881 mol, 9-10% aqueous solution) was added dropwise to compound Tr-3 (90 g, 0.294 mol) and RuCl3, xH₂ O (1.22 g, 0.0058 mol) in vigorously stirred solution in water (ml pH=8 room temperature). The pH was kept at 8 by adding 1 M NaOH solution. The reaction mixture was stirred at room temperature for 3 hours, and then heated at 35°C for 12 hours. After the reaction was complete (TLC), at 0 °C, 1.5 N HCl was added to the reaction mixture until pH 6 was reached, followed by the addition of EtOAc (1 L). Wash the organic phase with brine (2×100 mL) and dry (Na₂ SO₄ ), filtered and concentrated. The resulting crude product was purified by silica gel column chromatography on 230-400 silica with 20% EtOAc/petroleum ether to obtain compounds 4a+4b as a mixture of isomers. Subsequently, the isomers were separated by column chromatography on silica 230-400 using 0.9% MeOH/DCM and 0.1% AcOH as the dissolving agent to obtain 2R isomers (20 g, 28%).step 5) Acetic acid (2S)-2-(( Benzyloxy ) methyl )-1,3- Dioxolane -4- Ester (Tr-5) Pyridine (13.2 ml) and lead acetate (79.8 g, 0.180 mol) were added to a solution of compound Tr-4a (33 g, 0138 mol) in acetonitrile (660 mL) and the mixture was stirred at room temperature for 16 hours . After the reaction was complete (TLC), the reaction mixture was filtered, the filtrate was concentrated and the residue was dissolved in EtOAc (500 mL), washed with water (100 mL) and saturated sodium chloride solution (100 mL) and washed with Na₂ SO₄ dry. After removing the solvent, the crude product was purified by column chromatography on 60-120 silica using a 12-15% EtOAc/petroleum ether gradient to give the title compound (16 g, 47%) as a liquid.step 6) Acetic acid (2S)-2-( Hydroxymethyl )-1,3- Dioxolane -4- Ester (Tr-6) Pd/C (3.2 g, 20% w/w) was added to a stirred solution of compound Tr-5 (16 g) in anhydrous methanol (160 mL), and the reaction mixture was hydrogenated for 3 hours. After the reaction was complete (TLC), the reaction mixture was filtered through celite. The filtrate was concentrated under reduced pressure and the resulting crude title compound (10 g, 97%) was directly used in the next step.step 7) Acetic acid ((2S)-4- Acetoxy -1,3- Dioxolane -2- base ) Methyl ester (Tr-7) At 0°C, acetic anhydride (8.22 ml, 0.080 mol) was added to a stirred solution of compound Tr-6 (5.74 g, 0.0354 mol) in pyridine (107 ml) and the reaction mixture was stirred at room temperature for 16 hour. After the reaction was complete (TLC), the reaction mixture was quenched with dilute HCl (10 mL) and extracted into EtOAc (100 mL). Separate the organic phase and dry (Na₂ SO₄ ), filtered and concentrated. The resulting crude product was purified by column chromatography on 230-400 silica using 10-15% EtOAc/petroleum ether gradient dissolution to obtain the title compound (4.97 g, 68%) as a liquid.step 8) Acetic acid ((2S,4S)-4-(4-( Benzylamino )-2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methyl ester (Tr-8a) A mixture of N-benzyl cytosine (12.1 g, 56.3 mmol), ammonium sulfate (catalytic amount) and hexamethyldisilazane (HMDS) (67.4 ml, 418 mmol) was refluxed for 1 hour. At 40°C, HMDS was removed under reduced pressure and the residue was dissolved in anhydrous 1,2-dichloroethane (57 ml) and compound Tr-7 (5.7 g, 27.9 mmol) was added in anhydrous 1,2- A solution in dichloroethane (57 ml), followed by the dropwise addition of TMSOTf (10.2 ml, 45.7 mmol). The reaction mixture was stirred at room temperature for 1 hour, and then NaHCO was added₃ The aqueous solution and the mixture was stirred for 30 minutes. The resulting solid was filtered through Celite and the filtrate was dissolved in EtOAc (200 mL), washed with water (50 mL) and dried (Na₂ SO₄ ). After removing the solvent under reduced pressure, the crude product was purified by column chromatography on 230-400 silica using a 10-15% EtOAc/petroleum ether gradient to obtain a mixture of mutaisomers. The mixture of isomers was further separated by SFC purification to obtain the title compound (3 g, 30%) as a white solid.step 9) 4- Amine -1-((2S,4S)-2-( Hydroxymethyl )-1,3- Dioxolane -4- base ) Pyrimidine -2(1H)- ketone (Tr-9) At room temperature, a mixture of compound Tr-8a (3 g) and saturated methanolic ammonia solution (180 ml) was stirred in a sealed tube for 16 hours. After the reaction was complete (TLC), the solvent was removed under reduced pressure and the crude product was purified by column chromatography on 230-400 silica using 10-13% MeOH/DCM gradient dissolution to obtain the title compound as a solid (1.5 g, 85%).¹ H NMR 400 MHz DMSO-d₆ δ: 3.63-3.65 (2H), 4.04-4.07 (2H), 4.92-4.94 (1H), 5.18-5.21 (1H), 5.72-5.74 (1H), 6.16-6.18 (1H), 7.14 (1H), 7.26 (1H), 7.80-7.82 (1H).5-F- Preparation of Qusatabin

step 1) benzoic acid ((2S,4R)-4-(4- Benzoamide -5- fluorine -2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methyl ester (5-F-Tr-1a) and benzoic acid ((2S,4S)-4-(4- Benzoamide -5- fluorine -2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methyl ester (5-F-Tr-1b) A mixture of 5-fluorobenzyl cytosine (9.1 g, 39.5 mmol), ammonium sulfate (catalytic amount) and hexamethyldisilazane (140 ml) was refluxed for 14 hours. At 40°C, HMDS was removed under reduced pressure and the residue was dissolved in anhydrous 1,2-dichloroethane (50 ml) and the compound benzoic acid ((2S)-4-acetoxy-1, A solution of 3-dioxolane-2-yl) methyl ester (7 g, 26.30 mmol) in anhydrous 1,2-dichloroethane (50 ml) was then added dropwise with TMS-OTf (11.6 g, 52.6 mmol). The reaction mixture was allowed to stir at room temperature for 2 hours, then NaHCO₃ The aqueous solution was added to the reaction mixture and the mixture was stirred for another 30 minutes. The resulting solid was filtered through celite and the filtrate was dissolved in EtOAc (500 mL), washed with water (50 mL) and dried (Na₂ SO₄ ). The solvent was removed under reduced pressure and the crude product was purified by column chromatography on 230-400 silica using a 50-60% EtOAc/petroleum ether gradient to give the pure title compound as a solid (1.7g, 18% ).step twenty four- Amine -5- fluorine -1-((2S,4S)-2-( Hydroxymethyl )-1,3- Dioxolane -4- base ) Pyrimidine -2(1H)- ketone (5-F-Tr) At room temperature, a mixture of compound 5-F-Tr-1b (1.7 g) and saturated methanolic ammonia solution (34 ml) was stirred in a sealed tube for 16 hours, then the solvent was removed under reduced pressure and the crude product was passed through the tube Column chromatography was purified on a 230-400 silica using a 5% MeOH/DCM gradient to give the title compound as a solid (0.8 g, 68%). The following phenols were prepared and used to prepare intermediates of the compounds of this invention.phenol 1

step a) 1-(3-(( Tert-butyldimethylsilyl ) Oxygen ) Phenyl ) Ketone (Ph1-a) Imidazole (4.46 g, 65.5 mmol) was added to a solution of 3-hydroxyacetophenone (4.46 g, 32.8 mmol) in DMF (6 mL). After 5 minutes, a solution of TBDMS-Cl (4.69 g, 31.1 mmol) in DMF (4 mL) was added. The reaction mixture was allowed to stir at room temperature for 90 minutes, then poured into hexane (200 mL) containing 5% EtOAc and using 1M HCl (60 mL), water (60 mL), saturated sodium bicarbonate (2×60 mL), water (60 mL) and brine (60 mL). Organic layer via Na₂ SO₄ Dry, filter and concentrate and the resulting residue was purified by silica gel flash chromatography using hexane/EtOAc dissolution to obtain the title compound (5.7 g, 69%).step b) Tert-butyldimethyl (3-( C -1- Ene -2- base ) Phenoxy ) Silane (Ph1-b) Under nitrogen, methyl (triphenylphosphonium) bromide (10.2 g, 28.4 mmol) was suspended in anhydrous THF (30 mL) and the suspension was cooled to 0°C. N-Butyllithium (17.8 mL, 28.4 mmol) was added dropwise to the mixture and the resulting solution was stirred at room temperature for 30 minutes. Ph1-a (5.7 g, 22.8 mmol) was added to the mixture and the reaction was continued at room temperature for 60 minutes. The reaction was quenched with aqueous sodium bicarbonate solution and extracted with ether (50 mL). The organic layer was washed with sodium bicarbonate solution and dried (Na₂ SO₄ ), filtered and concentrated. The obtained residue was purified by silica gel plug with hexane dissolution to obtain the title compound (3.9 g, 69%).step c) Tert-butyldimethyl (3-(1- Methylcyclopropyl ) Phenoxy ) Silane (Ph1-c) Under nitrogen, within 10 minutes, a solution of diethylzinc in hexane (439.2 mmol) was added dropwise to the olefin Ph1-b (3.9 g, 15.7 mmol) in 1,2-dichloroethane (60 mL) In the cooling (0 ℃) solution. Diiodomethane (6.32 mL, 78.5 mmol) was added dropwise and the resulting mixture was stirred at 0 °C for 30 minutes and then allowed to reach room temperature overnight. The mixture was poured into an ice-cold solution of ammonium chloride and extracted with ether. The organic layer was washed with saturated sodium bicarbonate and dried (Na₂ SO₄ ), filtered and concentrated. The crude product was dissolved in hexane and the remaining diiodomethane was discarded. The hexane layer was concentrated to a crude product, which was used in the next step without further purification.step d) 3-(1- Methylcyclopropyl ) phenol ( phenol 1) Ph1-c (3.45 g, 13.1 mmol) was dissolved in a 1 M solution of tetrabutylammonium fluoride in THF (20 mL, 20 mmol) and the resulting solution was stirred at room temperature overnight. The reaction was quenched with 1M HCl (50 mL) and extracted with ethyl acetate (100 mL). The organic layer was washed with brine (2×50 mL) and dried (Na₂ SO₄ ), filtered and concentrated. The residue was purified by silica gel flash chromatography using a mixture of 2-propanol, EtOAc, and hexane to dissolve to obtain the title compound (0.56 g, 29%). MS 147.1 [M-H]^- .phenol 2

The title compound was prepared from 4-hydroxyacetophenone (6.0 g, 44.1 mmol) using the method described for the preparation of phenol 1. The yield is 53%.phenol 3

step a) 1-(3-( Benzyloxy ) Phenyl ) Cyclopentanol (Ph3-a) Iodine heated with magnesium was added to a suspension of magnesium shavings (1.29 g, 52.8 mmol) in anhydrous THF (50 mL). The mixture was refluxed and about 5% 3-bromophenol (13.9 g, 52.8 mmol) solution was added. When the reaction started, the bromide solution was added dropwise and then the mixture was refluxed for more than one hour. The mixture was cooled to about 5°C and a solution of cyclopentanone (4.44 g, 52.8 mmol) in THF (50 mL) was added dropwise. The mixture was stirred at room temperature for 72 hours, and then the reaction was quenched with a cooled saturated ammonium chloride solution and extracted with diethyl ether (×3). The organic phase was washed with brine and dried (Na₂ SO₄ ), filtered and concentrated. The resulting product was purified by silica gel chromatography (isohexane/EtOAc) to obtain the title compound (8.5 g, 54%).step b) 1-( Benzyloxy )-3-( Cyclopenta -1- Ene -1- base ) benzene (Ph3-b) P-Toluenesulfonic acid was added to a solution of Ph3-a (8.4 g, 28.2 mmol) in benzene (100 mL). The mixture was refluxed with a DMF trap for 3 hours, then cooled to rt, diluted with ether and washed with saturated sodium bicarbonate solution and brine. Dry organic phase (Na₂ SO₄ ), filtered and concentrated. The product was purified by silica gel chromatography (isohexane/EtOAc) to obtain the title compound (6.45 g, 91%). MS 249.4 [M-H]^- .step c) 3- Cyclopentylphenol ( phenol 3) Ph3-b (6.4 g, 26 mmol) in EtOAc (75 mL) and EtOH (75 in a Parr) at 22°C and 40 PSI in the presence of 10% Pd/carbon (1.5 g) mL) The solution was hydrogenated overnight. The catalyst was filtered off and washed with EtOAc and EtOH. The solvent was evaporated under reduced pressure and the product was purified by silica gel chromatography (isohexane/EtOAc) to give the title compound (3.6 g, 82%). MS 161.2 [M-H]^- .phenol 4

step a) Tertiary butyl (3- Cyclopropylphenoxy ) Dimethylsilane (Ph4-a) At 110 °C, (3-bromophenoxy) (third butyl) dimethyl silane (5.46 g, 19 mmol), cyclopropyl

Acid (2.12 g, 24.7 mmol), tripotassium phosphate (14.1 g, 66.5 mmol), tricyclohexylphosphine (0.53 g, 1.9 mmol) and Pd(OAc)₂ A suspension of (0.21 g, 0.95 mmol) in toluene (80 mL) and water (4 mL) was stirred overnight. The slurry was diluted with ether and washed with water and brine. Dry organic phase (MgSO₄ ), dried and concentrated. The crude product was purified by flash column chromatography (EtOAc/hexane) to give the title compound (1.94 g, 41%).step b) 3- Cyclopropylphenol ( phenol 4) 1 M tetrabutylammonium fluoride (10.1 ml, 10.1 mmol) was added to a solution of Ph4-a (1.94 g, 7,81 mmol) in THF (25 ml). The solution was stirred for 2 hours, then the solvent was evaporated and the residue was dissolved in EtOAc and concentrated NH₄ The Cl (aqueous) was washed twice and once with brine. Dry organic phase (MgSO₄ ), filtered and concentrated. The crude product was purified by flash column chromatography (hexane/ethyl acetate 9:1, containing 1% isopropanol) to obtain the slightly impure title compound (1.24 g, 119%).phenol 5

step a) 2-(4- Bromophenoxy ) Tetrahydro -2H- Piperan (Ph5-a) Dissolve 4-bromophenol (3.75 g, 21.7 mmol) in 3,4-dihydro-2H-piperan (16 ml, 175 mmol), add a catalytic amount of p-toluenesulfonic acid (15 mg, 0.09 mmol) and The mixture was stirred for 45 minutes at 22°C. The mixture was diluted with ether and washed with 1 M NaOH (aqueous solution)×2, water, and dried (Na₂ SO₄ ) And concentrated to give the title compound (5.57 g, 99%).step b) 2-(4- Cyclopropylphenoxy ) Tetrahydro -2H- Piperan (Ph5-b) Within 15 minutes, add 0.5 M cyclopropylmagnesium bromide in THF (6.5 ml, 3.25 mmol) to Ph5-a (552.5 mg, 2.15 mmol), ZnBr (144 mg, 0.64 mmol), tri-tert-butyl Phosphine tetrafluoroborate (35.6 mg, 0.12 mmol) and Pd(OAc)₂ (29.5 mg, 0.13 mmol) in THF (4 ml). At 22°C, the mixture was stirred for 90 minutes, then cooled on an ice bath and ice water (10 ml) was added. The mixture was extracted with EtOAc×3 and the extract was washed with brine, then dried (Na₂ SO₄ ), filtered and concentrated. The residue was purified by silica gel column chromatography (petroleum ether/EtOAc) to obtain the title compound (292 mg, 62%).step c) 4- Cyclopropylphenol ( phenol 5) P-Toluenesulfonic acid monohydrate (18.9 mg, 0.1 mmol) was added to a solution of Ph5-b (2.28 g, 10.45 mmol) in MeOH (15 ml). At 120°C, the mixture was heated in a microwave reactor for 5 minutes, then concentrated and purified by silica gel column chromatography (petroleum ether/EtOAc). The obtained solid was crystallized from petroleum ether to obtain the title compound (1.08 g, 77%).phenol 6

step a) 1-(3- Methoxyphenyl ) Cyclobutanol (Ph6-a) Between 0°C and 10°C, a 1 M solution of 3-methoxyphenylmagnesium bromide in THF (2.11 g, 99.8 mmol) was added dropwise to cyclobutanone (6.66 g, 95 mmol) in ether (65 mL) in the stirred solution. The mixture was stirred at 0-10°C for 3 hours, and then the mixture was added to ice-cold saturated NH₄ Cl solution (300 mL) and water (300 mL). The mixture was stirred for 10 minutes and then extracted three times with ether. Dry organic phase (Na₂ SO₄ ), filtered and concentrated. The obtained crude product was purified by silica gel chromatography (isohexane/EtOAc) to obtain the title compound (16.9 g, 86%).step b) 1- Cyclobutyl -3- Methoxybenzene (Ph6-b) 10% Pd/carbon (2.5 g) was added to a solution of Ph6-a (15.4 g, 86.1 mmol) in ethanol (200 mL), and the mixture was hydrogenated at 60 psi in a Parr device. After 18 hours, another 10% pd/carbon (1.5 g) was added and the mixture was hydrogenated at 60 psi for an additional 18 hours. The catalyst was filtered off and washed with EtOH and EtOAc. The solution was concentrated under reduced pressure, and the crude product was separated by silica gel chromatography (isohexane/EtOAc) to obtain the title compound (14.0 g, 77%).step c) 3- Cyclobutylphenol ( phenol 6) At 0 °C, a solution of 1 M boron tribromide (18.1 g, 72.2 mmol) in DCM was added dropwise to a solution of Ph6-b (10.6 g, 65.6 mmol) in anhydrous DCM (65 mL). The mixture was stirred at -5°C for 2.5 hours, then saturated NH with cooling₄ The Cl solution was quenched and extracted three times with DCM. Dry organic phase (Na₂ SO₄ ), filtered and concentrated. The obtained crude product was purified by silica gel chromatography (isohexane/EtOAc) to obtain the title compound (9.73 g, 88%).phenol 7

step a) 1-(4-( Benzyloxy ) Phenyl ) Cyclobutanol (Ph7-a) Under reflux, a solution of 1-(benzyloxy)-4-bromobenzene (2.63 g, 100 mmol) in diethyl ether:THF 1:1 (100 mL) was added dropwise to magnesium shavings in about 1 h (2.43 g) and trace iodine in suspension in ether (50 mL). When the addition was complete, the mixture was refluxed for four hours and then cooled to about 0°C. Anhydrous THF (50 ml) was added, followed by slowly adding a solution of cyclobutanone (7.01 g, 100 mmol) in diethyl ether (50 mL) and the mixture was allowed to reach room temperature. After stirring for two hours, add cooling saturated NH₄ Cl solution (500 ml) and stir the mixture for 15 minutes, then extract twice with EtOAc. The organic phase was washed with brine, dried with sodium sulfate and evaporated under reduced pressure. The product was purified by silica gel column chromatography to obtain the title compound (12.5 g, 42%).step b) 4- Cyclobutylphenol ( phenol 7) Under argon, add 10% Pd/carbon (2.55 g, 21.5 mmol) to a solution of Ph7-a (12.4 g, 41.4 mmol) in anhydrous EtOH (110 mL) at 45 psi at room temperature Next, the mixture was hydrogenated for 18 hours. The catalyst was filtered off, washed with ethanol and the solution was concentrated. The product was purified by silica gel chromatography (isohexane-EtOAc). The appropriate fractions were combined and concentrated and the residue was crystallized from petroleum ether to give the title compound (3.15 g, 51%).phenol 8

4-(1- Methylcyclopentyl ) phenol ( phenol 8) Within 30 minutes, a solution of 1-methylcyclopentanol (2.00 g, 20.0 mmol) and phenol (2.07 g, 22.0 mmol) in pentane (50 mL) was added dropwise to fresh AlCl₃ (1.33 g, 10 mmol) in suspension in pentane (100 mL). In N₂ At room temperature, the resulting mixture was stirred for 72 hours, and then the reaction mixture was poured into water/ice and HCl (12 M, 20 mmol, 1.66 mL). Wash the organic phase with water (50 mL) and brine (50 mL) and dry (Na₂ SO₄ ), filtered and concentrated. The crude product was purified by silica gel column chromatography (MeOH-DCM) to obtain the title compound (426 mg, 12%).phenol 9

step a) 2-(4- bromine -3- Methylphenoxy ) Tetrahydro -2H- Piperan (Ph9-a) Add pTs (16 mg, 0.086 mmol) to 4-bromo-3-methylphenol (4.0 g, 21.4 mmol) in 3,4-dihydro-2-H-piperan (16 mL, 175 mmol) Solution. The reaction mixture was stirred at room temperature for 1 hour, then diluted with ether and washed with 1M NaOH (aq) and water. Dry organic phase (Na₂ SO₄ ), filtered and concentrated. The crude product was purified by silica gel column chromatography (EtOAc/heptane) to give the title compound (3.32 g, 57%).step b) 2-(4- Cyclopropyl -3- Methylphenoxy ) Tetrahydro -2H- Piperan (Ph9-b) Ph9-a (3.12 g, 11.5 mmol), ZnBr₂ (2.59 g, 11.5 mmol), tri-tert-butylphosphine tetrafluoroborate (0.2 g, 0.69 mmol) and Pd(OAc)₂ (258 mg, 1.15 mmol) placed in a flask and using N₂ Rinse the flask several times. THF (10 mL) was added with stirring, and then 0.5 M cyclopropylmagnesium bromide in THF solution (35 mL, 17.4 mmol) was added dropwise within 5 minutes. The mixture was stirred at room temperature, then filtered through a plug of celite and dissolved with MeOH. The solution was concentrated and the crude product was purified by silica gel column chromatography (EtOAc/heptane) to give the title compound (1.69 g, 57%).step c) 4- Cyclopropyl -3- Methylphenol ( phenol 9) Dissolve Ph9-b (1.70 g, 7.30 mmol) in MeOH (20 ml) and add pTsxH₂ O (318 mg, 1.67 mmol). The mixture was stirred at 22°C for 30 minutes and then concentrated. The crude product was purified by column chromatography (EtOAc/heptane) to give the title compound (704 mg, 65%).phenol 10

step a) 4- Cyclopropyl -1- Methoxy -2- Methylbenzene (Ph10-a) According to the procedure described in Ph9 step b, 4-bromo-1-methoxy-2-methylbenzene (4.39 g, 21.9 mmol) was reacted with cyclopropylmagnesium bromide to give the title compound (1.54 g, 43% ).step b) 4- Cyclopropyl -2- Methylphenol ( phenol 10) In N₂ At 0 ℃, the BBr₃ (5 mL, 5 mmol) was added to a solution of Ph10-a (1.54 g, 9.49 mmol) in DCM (7.5 mL). The reaction was allowed to stir for 2 hours, then quenched with MeOH (3 mL) and concentrated. The crude product was dissolved in EtOAc and washed with brine. Dry organic phase (Na₂ SO₄ ), filtered and concentrated. The crude product was purified by silica gel column chromatography to obtain the title compound (826 mg, 59%). MS 147.11 [M-H]^- .phenol 11

4- Cyclopropyl -3- Methoxyphenol ( phenol 11) The title compound was prepared from 4-bromo-3-methoxyphenol (1.11 g, 5.49 mmol) according to the procedure described for the preparation of phenol 9. The yield is 40%.phenol 12

step a) 3-( Dimethylamino )-1-(3- Hydroxyphenyl ) C -1- ketone (Ph12-a) Add a few drops of HCl to 3-hydroxyacetophenone (4.08 g, 30 mmol), paraformaldehyde (4.05 g, 45 mmol) and dimethylamine hydrochloride (2.69 g, 33 mmol) in anhydrous EtOH (100 mL ) And reflux the reaction mixture for 18 hours. Additional dimethylamine hydrochloride (0.55 equivalents, 1.22 g), paraformaldehyde (0.5 equivalents, 1.35 g) and HCl (0.5 mL) were added and the reaction mixture was refluxed for another 4 hours, then cooled to room temperature. The deposited white solid was collected and washed with cold EtOH (50 mL) and cold acetone (10 mL), followed by freeze drying to give the title compound (2.59 g, 38%), which was used in the next step without further purification.step b) Cyclopropyl (3- Hydroxyphenyl ) Ketone ( phenol 12) At room temperature, NaH (60% mineral oil dispersion) (1.13 g, 28.2 mmol) was added portionwise to stirred suspension of trimethyl iodide oxide (6.20 g, 28.2 mmol) in DMSO (100 mL) In the liquid. After 1 hour, with stirring and cooling, solid Ph12-a (2.59 g, 11.3 mmol) was added in portions. The reaction mixture was stirred at room temperature for 40 hours, then poured into cold water (200 mL) and extracted with DCM (3×100 mL). Use saturated NH₄ The organic phase was washed with Cl aqueous solution (2×100 mL) and dried (Na₂ SO₄ ), filtered and concentrated. The obtained crude product was purified by silica gel column chromatography (MeOH/DCM) to obtain the title compound (883 mg, 48%).phenol 13

step a) Cyclopropyl (4- Hydroxyphenyl ) Ketone (Ph13) Within about 30 minutes, p-hydroxy-γ-chlorobutyrophenone (4.95 g) was added portionwise to the NaOH solution (8 mL, aqueous solution, 50% w/w), followed by NaOH (35 mL, aqueous solution, 25% w/w), followed by one-time addition of p-hydroxy-γ-chlorobutyrophenone (4.95 g). The temperature was lowered to 140°C and NaOH (8 g) was added. After 90 minutes, add H₂ O (10 mL), and after another 60 minutes, the reaction mixture was cooled, using H₂ O was diluted and neutralized to a pH of about 7 with HOAc (about 27-30 ml). Filter the precipitate formed and use H₂ O washed and dried in vacuo. At 40 ℃, make the solid in CHCl₃ (200 ml) for 10 minutes, then overnight at room temperature. The slurry was heated to 40°C within 30 minutes and then filtered. Dry filtrate (MgSO₄ ), filtered and concentrated to about 70 ml. Hexane is added and an oil is formed, which eventually becomes crystals. Filter the slurry using CHCl₃ /Hexane was washed and dried to obtain the title compound (4.15 g, 51%).phenol 14

step a) 3-(1- Hydroxyl -2,2- Dimethylpropyl ) phenol (Ph14-a) Within 30 minutes, t.Bu-MgBr (1.5 equivalents) was added dropwise to a cold (-10°C) mixture of 3-hydroxybenzaldehyde (2.00 g, 16.4 mmol) in diethyl ether (20 mL). During the addition, THF (20 mL) was added. The mixture was brought to 23°C and stirred for 6 hours. Add more t.Bu-MgBr (0.7 equiv) and let the mixture stir overnight, then cool and use saturated NH₄ The aqueous Cl solution quenched the reaction. EtOAc was added to the mixture, followed by 1 M aqueous HCl until a homogeneous mixture was obtained. Separate the phases and wash the organic phase with brine and dry (Na₂ SO₄ ), filtered and concentrated. The obtained crude product was purified by column chromatography to obtain the title compound (1.1 g, 37%).step b) 1-(3- Hydroxyphenyl )-2,2- Dimethylpropane -1- ketone (Ph14) 3 Å MS and pyridinium chlorochromate (PCC) (1.97 g, 9.15 mmol) and anhydrous DCM (5 mL) were successively added to an oven-dried round bottom flask. The mixture was stirred at 20°C for 5 minutes, then a mixture of AA8019 (1.10 g, 6.10 mmol) in DCM (5 mL) was slowly added. After complete oxidation, the mixture was filtered through a pad of diatomaceous earth, and the pad was washed with ether. Concentrate the filtrate. The crude product was purified by column chromatography to obtain the title compound (402 mg, 37%). MS 179.25 [M+H]⁺ .phenol 15

1-(4- Hydroxyphenyl )-2,2- Dimethylpropane -1- ketone (Ph15) 4-Hydroxybenzaldehyde (3 g, 24.6 mmol) was reacted according to the procedure described in the preparation of phenol 14, to give the title compound (538 mg, 17%).Amino acid 1

step a) (S)-(S)-2-(( Third butoxycarbonyl ) Amine ) Second butyl propionate (AA1-a) L-Boc-alanine (2.18 g, 11.5 mmol) was dissolved in anhydrous DCM (40 mL) and alcohol (R)-butan-2-ol (938 mg, 12.6 mmol) was added. The mixture was cooled to about 5°C and EDC (3.31 g, 17.2 mmol) was added in one portion, followed by DMAP (140 mg, 1.15 mmol) in portions. The mixture was kept at room temperature and stirred overnight, then diluted with ethyl acetate (about 300 ml) and washed the organic phase three times with saturated sodium bicarbonate solution and once with brine. The organic phase was dried over sodium sulfate and concentrated under reduced pressure. The product was separated by silica gel chromatography using isohexane and 10% ethyl acetate dissolution to obtain the title compound (2.78 g, 98%).step b) (S)-(S)-2- Second butyl aminopropionate (AA1-b) At 65 °C, a mixture of AA1-a (2.77 g, 11.3 mmol) and p-toluenesulfonic acid monohydrate (2.15 g, 11.3 mmol) in EtOAc (45 mL) was stirred for 16 hours, and then concentrated under reduced pressure . The obtained residue was crystallized from diethyl ether to obtain the title compound (3.20 g, 89%).Amino acid 2

(S)-(R)-2- Alanine -2- Ester (AA2) According to the procedure described in the preparation of AA1, but using (R)-pent-2-ol instead of (R)-butan-2-ol, the title compound (4.6 g) was obtained.Amino acid 3

(S)-(S)-2- Alanine -2- Ester (AA3) According to the procedure described in the preparation of AA1, but using (S)-pentan-2-ol instead of (R)-butan-2-ol, the title compound (8.3 g) was obtained. The following intermediates are prepared and can be used to prepare the compounds of the present invention:Intermediate 1

step a) (R)-2-(( Third butoxycarbonyl ) Amine ) Propionic acid 4- Fluorobenzyl ester (I-1a) Dissolve Boc-L-AlaOH (19.92 mmol), DMAP (1.99 mmol) and (4-fluorophenyl)methanol (23.9 mmol) in CH₂ Cl₂ (100 mL). Successively add triethylamine (23.9 mmol) and EDCI (23.9 mmol) to this solution and add₂ At room temperature, the resulting reaction mixture was stirred overnight. Use CH₂ Cl₂ (100 mL) Dilute the reaction mixture using saturated NaHCO₃ Aqueous solution (2×50 mL), saturated aqueous NaCl solution (2×50 mL), dried (Na₂ SO₄ ) And concentrated. The obtained residue was purified by silica gel column chromatography using n-hexane-EtOAc (95:5 to 60:40) dissociation to obtain the title compound (4.44 g) as a white waxy solid. MS: 296 [M-H]^- .step b) (R)-2- Alanine 4- Fluorobenzyl ester (I-1b) Compound I-1a (14.93 mmol) was dissolved in 4M HCl/dioxane (40 mL) and stirred at room temperature for 30 minutes and evaporated to dryness to give the hydrochloride salt of the title compound (3.4 g) as a white powder ). MS: 198 [M+H]⁺ .step c) (2R)-2-(( chlorine ( Phenoxy ) Phosphoryl ) Amine ) Propionic acid 4- Fluorobenzyl ester (I-1) At -78 ℃, the PhOPOCl₂ (4.28 mmol) was added dropwise to the CH of compound 1-5b (4.28 mmol)₂ Cl₂ To the solution, triethylamine (8.56 mmol) was then added dropwise. Under Ar, at -78°C, the resulting reaction mixture was stirred and kept at room temperature overnight. The reaction mixture was evaporated on silica gel and purified by chromatography (n-hexane/EtOAc (88:12)-(0:100)) to obtain the title compound (769 mg).³¹ P-NMR (CDCl₃ ) δ: 7.85 (s) and 7.54 (s) (R_P And S_P Diastereomer).Intermediate 2

step a) (S)-(R)-2-(( Third butoxycarbonyl ) Amine ) Second butyl propionate (I-2a) L-Boc-alanine (2.18 g, 11.5 mmol) was dissolved in anhydrous DCM (40 mL) and alcohol (R)-butan-2-ol (938 mg, 12.6 mmol) was added. The mixture was cooled to about 5°C and EDC (3.31 g, 17.2 mmol) was added in one portion, followed by DMAP (140 mg, 1.15 mmol) in portions. The mixture was kept at room temperature and stirred overnight, then diluted with ethyl acetate (about 300 ml) and washed the organic phase three times with saturated sodium bicarbonate solution and once with brine. The organic phase was dried over sodium sulfate and concentrated under reduced pressure. The product was separated by silica gel chromatography using isohexane and 10% ethyl acetate dissolution to obtain the title compound (2.78 g, 98%).step b) (S)-(R)-2- Second butyl aminopropionate (I-2b) At 65°C, a mixture of I-10a (2.77 g, 11.3 mmol) and p-toluenesulfonic acid monohydrate (2.15 g, 11.3 mmol) in EtOAc (45 mL) was stirred for 16 hours, and then concentrated under reduced pressure . The obtained residue was crystallized from diethyl ether to obtain the title compound (3.20 g, 89%).step c) (2S)-(R)-2-(((4- Nitrophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Second butyl propionate (I-2) Under nitrogen, at -30°C, phenyl dichlorophosphate (1 equivalent) was added to a solution of compound I-10b (3.15 g, 9.92 mmol) in DCM (75 ml), followed by the dropwise addition of triethyl Amine (2 equivalents). The mixture was kept at room temperature and stirred overnight, then cooled to about 5°C and 4-nitrophenol (1 equivalent, 15 mmol) was added as a solid, followed by dropwise addition of triethylamine (1 eq g, 15 mmol), and The mixture was stirred at room temperature for 4 hours, and then concentrated under reduced pressure, diluted with ethyl acetate (40 ml) and diethyl ether (40 ml) and kept at room temperature overnight. The triethylamine-HCl salt was filtered off and the filtrate was concentrated under reduced pressure. The obtained residue was purified by silica gel column chromatography using isohexane-ethyl acetate dissociation to obtain the title compound (4.19 g, 79%). The following compounds are prepared using the appropriate alcohol according to the procedure described in Preparation I-2:

Intermediate 6 , Diastereomer -1 and -2 The two diastereomers of compound I-6 were separated by SFC to obtain I-6-diastereomer-1 and I-6-diastereomer-2.Intermediate 7

step a) (S)-2- Aminopropionate cyclooctyl ester (I-7a) P-Toluenesulfonic acid monohydrate (3.6 g, 19.1 mmol) was added to a slurry of L-alanine (1.7 g, 19.1 mmol) and cyclooctanol (25 ml, 191 mmol) in toluene (100 ml). The reaction mixture was heated at reflux temperature for 25 hours and water was removed from the reaction using a Dean-Stark separator. The mixture was concentrated under reduced pressure and the residue was kept under vacuum overnight. Diethyl ether (100 ml) was added to the residue (27 g). The white precipitate was collected by filtration, washed with ether (3×50 ml) and dried under vacuum to give the title compound (4.84 g, 68%).step b) (2S)-2-(((4- Nitrophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Cyclooctyl propionate (I-7) Compound I-7a was reacted according to the method described in step c of Preparation I-2 to obtain the title compound (4.7 g, 76%).Intermediate 8

(2S)-2-(((4- Nitrophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Cycloheptyl propionate (I-22) Following the procedure described for the preparation of compound I-7, but using cycloheptanol (27 ml, 224 mmol) instead of cyclooctanol, the title compound (5.72 g, 55%) was obtained.Intermediate 9

Follow the procedure described in step c of Preparation I-2, but using (S)-2-aminopropionic acid cyclohexyl ester instead of (S)-2-aminopropionic acid 3,3-dimethylbutyl ester to give the title Compound (10.6 g, 82%).Intermediate 10

(S)-2-(( two (4- Nitrophenoxy ) Phosphoryl ) Amine ) Propionic acid 2- Ethyl butyl ester (I-10) (S)-2-Aminopropionic acid 2-ethylbutyl ester (5 g, 14.49 mmol) was added to bis(4-nitrophenyl) chlorophosphate (6.14 g, 17.1 mmol) in DCM (50 ml ), the mixture was cooled in an ice bath and Et was added dropwise₃ N (4.77 mL, 34.2 mmol). After 15 minutes, the cooling was removed and the reaction mixture was stirred at 23 °C until the reaction was complete according to TLC. Then diethyl ether was added, the mixture was filtered and the filtrate was concentrated and purified by silica gel column chromatography to obtain the title compound (2.05 g, 82%).Intermediate 11

step a) (S)-2- Isopropyl aminopropionate (I-11a) At 0 ℃, the SOCl₂ (29 mL, 400 mmol) was added dropwise to a suspension of L-alanine HCl salt (17.8 g, 200 mmol) in isopropanol (700 mL). The suspension was stirred at room temperature overnight, and then concentrated to give the title compound (29.2 g, 87%).step b) (2S)-2-((((((S)-1- Isopropoxy -1- Pendant -2- base ) Amine )(4- Nitrophenoxy ) Phosphoryl )- Amine ) Isopropyl propionate (I-11) At -60 °C, a solution of 4-nitrophenyl dichlorophosphate (1.8 g, 7 mmol) in DCM was added dropwise to amine I-11a (2.35 g, 14 mmol) and triethylamine (7.7 mL , 56 mmol) in DCM. The reaction mixture was kept at room temperature, stirred overnight, concentrated and then diluted with ethyl acetate and ether and kept at room temperature overnight. The triethylamine HCl salt was filtered off, the filtrate was concentrated under reduced pressure and the resulting residue was purified by silica gel chromatography using isohexane-ethyl acetate dissolution to obtain the title compound (1.6 g, 50%).Intermediate 12

step a) (S)-2-(( Third butoxycarbonyl ) Amine ) Neopentyl propionate (I-12a) At -5°C, EDAC and DMAP were added portionwise to a solution of Boc-alanine (18.9 g, 100 mmol) and neopentyl alcohol (13.0 mL, 120 mmol) in DCM (200 mL). The reaction mixture was kept at room temperature and stirred for 72 hours. Add EtOAc (700 mL) and use saturated NaHCO₃ The solution was washed three times with organic phase and once with brine, then concentrated. The obtained residue was purified by column chromatography using hexane-EtOAc 90/10 to 80/20 dissolution to obtain the title compound (21 g, 81%).step b) (S)-2- Neopentyl aminopropionate (I-12b) At -65°C, p-toluenesulfonic acid (15.6 g, 82.0 mmol) was added to a solution of Boc-protected amine I-12a (21.1 g, 82.0 mmol) in EtOAc (330 mL). The reaction mixture was allowed to stir at -65°C for 8 hours, then kept at room temperature overnight. Subsequently, the mixture was filtered and concentrated to obtain the title compound (21 g, 78%).(2S)-2-((((((S)-1-( Neopentyloxy )-1- Pendant -2- base ) Amine )(4- Nitrophenoxy )- Phosphoryl ) Amine ) Neopentyl propionate (I-12) At -50°C, 4-nitrophenol dichlorophosphate was added dropwise to a solution of amine I-12b (3.90 g, 24.5 mmol) in DCM (100 mL) within 1 hour. The reaction mixture was kept at room temperature, stirred overnight, concentrated and then diluted with ether and overnight at room temperature. The mixture was filtered, and the filtrate was concentrated under reduced pressure and the resulting residue was purified by silica gel chromatography using isohexane-ethyl acetate dissolution to obtain the title compound (4.8 g, 77%).Intermediate 32

(2S)-(R)-2-(((( Perfluorophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Second butyl propionate (I-32) At -70 ℃, under nitrogen, Et within 15 minutes₃ N (10.9 mL, 78.1 mmol) was added dropwise to the stirred solution of (S)-(R)-2-aminopropionic acid second butyl pTs salt (12.0 g, 37.7 mmol) in DCM (50 mL) in. Within 1 hour, a solution of phenyl dichlorophosphate (5.61 mL, 37.7 mmol) in DCM (50 mL) was added to the mixture. At -70°C, the reaction mixture was stirred for another 30 minutes, and then heated to 0°C within 2 hours and stirred for 1 hour. Within 20 minutes, combine pentafluorophenol (6.94 g, 37.7 mmol) and Et₃ A solution of N (5.73 mL, 41.1 mmol) in DCM (30 mL) was added to the mixture. The crude mixture was stirred at 0°C for 18 hours, and then concentrated. The residue was dissolved in THF (100 mL), the insoluble material was filtered off and washed with THF multiple times. The solvent was evaporated and the residue was triturated with tert-butyl methyl ether. The insoluble matter was filtered off and washed with tert-butyl methyl ether. The combined filtrate was concentrated and the crude solid was sonicated with n-hexane/EtOAc (80:20; 100 mL). The solid was filtered and washed with n-hexane/EtOAc (80:20) to give the pure phosphorus stereoisomer of the title compound (2.3 g, 13%) as a white solid.Intermediate 33

(2S)-2-(((( Perfluorophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Ethyl propionate (I-33) The pure phosphorus stereoisomer system of the title compound was prepared according to the method described for I-32, but starting with the HCl salt of ethyl (S)-2-aminopropionate (11.0 g, 71.1 mmol). The yield was 8.56 g, 27%.Intermediate 34

(2S)-2-(((( Perfluorophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Propionic acid 2- Ethyl butyl ester (I-34) The pure phosphorus stereoisomeric system of the title compound was prepared according to the method described for I-32, but starting with the pTs salt (18.8 g, 54.4 mmol) of (S)-2-aminopropionic acid 2-ethylbutyl ester Starting thing. The yield was 27.0 g, 99%. LC-MS 496.44 [M+H]⁺ .Intermediate 35

(2S)-2-(((( Perfluorophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Butyl propionate (I-35) Add phenyl dichlorophosphate (12.4 mL, 83.1 mmol) to (S)-2-aminopropionic acid butyl ester (26.4 g, 83.1 mmol) in dichloromethane (200 mL) in cold (-20°C) In the slurry. The mixture was stirred for 10 minutes, and then Et was added dropwise within 15 minutes₃ N (25.5 mL, 183 mmol). The mixture was stirred at -20°C for 1 hour, followed by 30 minutes at 0°C. The mixture was cooled in an ice bath and perfluorophenol (15.3 g, 0.08 mol) was added, followed by dropwise addition of Et₃ N (11.6 mL, 0.08 mol). The mixture was stirred overnight and slowly raised to 20°C. Diethyl ether was added and the mixture was filtered through diatomaceous earth, concentrated and purified by silica gel column chromatography using petroleum ether/EtOAc (9:1 -> 8:2) dissolution. The appropriate fractions were combined, concentrated and crystallized from petroleum ether/EtOAc (9:1) to give the pure phosphorus stereoisomer of the title compound (2.23 g, 5.8%) as a white solid.Intermediate 36

step a) L- Isopropyl Alanine Hydrochloride (I-36a) At -7 to 0°C, add thiosulfonyl chloride (80.2 g, 0.674 mol, 1.5 equivalents) to 2-propanol (400 mL) under cooling over a period of 30 minutes, followed by 0°C , L-alanine (40.0 g, 0.449 mol) was added. Connect a flow indicator and a scrubber with a mixture of 27.65% sodium hydroxide (228 g) and water (225 g) to the outlet. The reaction mixture was allowed to stir at 67°C for 2 hours, then at 70°C for 1 hour and at 20-25°C overnight. The reaction mixture was distilled off from the 60°C bath at 47-50°C under reduced pressure (250-50 mbar). When the distillation becomes very slow, add toluene (100 mL) to the residual oil and continue to distill from the 60°C bath at 48-51°C under reduced pressure (150-50 mbar) until it becomes Very slowly. Third butyl methyl ether (tBME) (400 mL) was added to the residual oil, and the two-phase system was seeded at 34-35°C with effective stirring. When crystallization was observed, the mixture was cooled to 23°C over a period of one hour, and the precipitate was separated by filtration. The filter cake was washed with tBME (100 mL) and dried to a constant weight without heating under reduced pressure to obtain the title compound (67.7 g, 90%) as a white solid.step b) (S)-2-(((S)-( Perfluorophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Isopropyl propionate (I-36) At 0 °C, under nitrogen, phenyl dichlorophosphate (62.88 g, 0.298 mol, 1.0 equiv) was added to isopropyl L-alanine hydrochloride (50.0 g, 0.298 mol) in DCM (310 mL) In the solution in, the addition was completed by washing with DCM (39 mL). The mixture was allowed to cool and over a period of 70 minutes, under cooling, triethylamine (63.35 g, 0.626 mol, 2.1 equivalents) was added, keeping the temperature not exceeding -14°C, and the addition was completed by washing with DCM (39 mL) . The mixture was stirred at -15 to -20°C for 1 hour, then heated to -8°C and over a period of 42 minutes, perfluorophenol (60.38 g, 0.328 mol, 1.1 equivalent) and triethylamine (33.19 g were added , 0.328 mol, 1.1 equiv) in DCM (78 mL), keeping the temperature not to exceed 0 °C, the addition was completed by washing with DCM (39 mL). The mixture was allowed to stir at 0°C for 1 hour, then at +5°C overnight. The precipitate formed was removed by filtration, and the filter cake was washed with DCM (95 mL). The combined filtrate was washed with water (2×190 mL) at 5°C. The organic phase was distilled under reduced pressure (650-600 mbar) at 32-38°C, and distillation continued until the remaining volume was about 170 mL. Some crystals were obtained. Ethyl acetate (385 mL) was added and the resulting clear solution was distilled at 43-45°C under reduced pressure (300-250 mbar). Continue distillation until the remaining volume is about 345 mL. The clear solution was cooled to 36°C, by adding (S)-2-(((S)-(perfluorophenoxy)) prepared according to J. Org. Chem., 2011, 76, 8311-8319 Seed crystals (20 mg) of phenoxy)phosphoryl)amino)propionic acid isopropyl propionate induce crystallization. Over a period of 1 hour, the mixture was cooled to 27°C, then n-heptane (770 mL) was added over a period of 47, and the mixture was stirred for another 37 minutes. Triethylamine (6.03 g, 0.2 equiv) was added and the mixture was stirred at 23-25°C overnight. The precipitate was separated by filtration. The filter cake was washed with ethyl acetate: n-heptane (1:9, 80 mL) and dried under reduced pressure (less than 0.1 mbar) to constant weight without heating to give the title compound (75.64) as white crystals g, 56%).¹ H NMR (CDCl₃ , 300 MHz) δ 7.38-7.32 (m, 2 H), 7.27-7.24 (m, 2 H), 7.23-7.19 (m, 1 H), 5.10-4.98 (m, 1 H), 4.20-4.08 (m , 1 H), 4.03-3.96 (m, 1 H), 1.46 (dd, 7.2, 0.6 Hz, 3 H), 1.26-1.23 (2xd, 6 H);¹³ CNMR (CDCl₃ , 100 MHz) δ 172.7 (d, J = 8.8 Hz), 150.4 (d, J = 7.1 Hz), 143.4-143.0 (m), 141.0-140.2 (m), 140.0-139.8 (m), 137.6-137.2 ( m), 136.8-136.2 (m), 130.0 (d, J = 0.82 Hz), 125.8 (d, J = 1.4 Hz), 120.3 (d, J = 5.0 Hz), 69.8, 50.6, (d, J = 1.9 Hz), 21.8 (d, J = 1.9 Hz), 21.2 (d, J = 4.4 Hz); The crystalline properties and NMR spectral data of the title compound are consistent with published information (J. Org. Chem., 2011, 76, 8311-8319), thus confirming the S stereochemistry of the phosphorus atom of the title compound.Intermediate 37

step a) (S)-2- Cyclohexyl propionate (I-37a) Acetyl chloride (4.2 mL, 59.3 mmol) was added dropwise to a stirred solution (50 ml) of cyclohexanol, followed by L-phenylalanine (4.0 g, 24.2 mmol). The reaction mixture was heated to 100° C. for 16 hours, and then concentrated under reduced pressure, triturated with ether/hexane (1:1) and dried to give the title compound (6 g, 88%) as a white solid. The compound was used in the next step without further purification.step b) (S)-2-(((S)-( Perfluorophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Cyclohexyl propionate (I-37) At -70°C, triethylamine (7.17 mL, 51.5 mmol) was added dropwise to a stirred solution of compound 1-37a (7.0 g, 24.6 mmol) in anhydrous DCM (42 mL) over 30 minutes, followed by A solution of phenyl dichlorophosphate (5.15 g, 34.5 mmol) in anhydrous DCM (21 mL) was added over 1 hour. The reaction mixture was stirred at -70°C for another 30 minutes, and then heated to 0°C over 2 hours and stirred for 1 hour. Over 1 hour, a solution of perfluorophenol (4.94 g, 26.8 mmol) and triethylamine (3.74 mL, 26.8 mmol) in anhydrous DCM (28 mL) was added to the mixture. The mixture was stirred at 0°C for 4 hours, followed by 5°C for 16 hours. The reaction mixture was filtered and the filtrate was concentrated under reduced pressure. The residual solid was dissolved in EtOAc (300 mL), washed with water (50 mL), dried and the solvent was removed under reduced pressure. The resulting solid was triturated with 20% EtOAc in hexane, filtered, washed with hexane and dried to give the title compound as a single diastereomer as a solid (3.0 g, 21%).Intermediate 38

(2S)-2-(((4- Nitrophenoxy )( Phenoxy ) Phosphoryl ) Amine ) Isopropyl propionate (I-38) At -78°C, a solution of phenol (1.86 g, 19.8 mmol) and triethylamine (3 mL, 21.8 mmol) in anhydrous DCM (50 mL) was added to 4-nitrodichlorophosphate over a period of 30 minutes Phenylphenyl ester (5 g, 19.8 mmol) in a stirred solution in anhydrous DCM (40 ml). The mixture was stirred at this temperature for 60 minutes, and then transferred to a compound containing compound (S)-2-aminopropyl propionate (3.3 g, 19.8 mmol) in anhydrous at -5°C over a period of 15 minutes. The solution in DCM (40 mL) was in another flask. At -5 °C, a second portion of TEA (6 mL, 43.3 mmol) was added to the mixture over a period of 20 minutes. The mixture was stirred at 0°C for 3 hours, and then the solvent was removed under reduced pressure. The residue was dissolved in EtOAc (200 mL) and washed with water (50 mL), washed with Na₂ SO₄ Dry and remove the solvent under reduced pressure to obtain the crude product as an oil, which is then purified by column chromatography using a 0-20% EtOAc/hexane gradient and 230-400 mesh silica gel to obtain a ratio of about 1:1 Mixture of diastereomers. The diastereomers were separated by SFC to obtain the title compound, Isomer 1 (1.5 g, 20%) and Isomer 2 (1.5 g, 18%) as solids. The compounds listed in Table 1 were prepared using the appropriate amino acid esters and phenol according to the procedure described in Preparation Intermediate 1-38, and the diastereomers were separated. Table 1

Examples 1

step a) Acetic acid ((2S,4S)-4-(2,4- Dioxo -3,4- Dihydropyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methyl ester (1a) At 125°C, a mixture of compound Tr-8 (0.15 g, 0.41 mmol), 1,2-dimethoxyethane (1.5 mL) and water (0.96 mL) was heated in a sealed tube for 48 hours. After the reaction was complete (TLC), the reaction mixture was cooled to room temperature and the solvent was removed under reduced pressure. The crude residue was purified by column chromatography on 230-400 silica gel using a 3-7% MeOH/DCM gradient to obtain compound 1a (0.08 g, 80%) as a solid and compound 1b (0.02 as a solid g).step b) 1-((2S,4S)-2-( Hydroxymethyl )-1,3- Dioxolane -4- base ) Pyrimidine -2,4(1H,3H)- Dione (1b) At room temperature, make saturated NH containing compound 1a (0.08 g, 0.31 mmol)₃ The MeOH solution (1.6 mL) was stirred in a sealed tube for 4 hours. After the reaction was complete (TLC), the solvent was removed under reduced pressure and the residue was purified by column chromatography on 60-120 silica gel using 5-7% MeOH/DCM to obtain the title compound (0.06 g, 90%).step c) (2S)-2-((((((2S,4S)-4-(2,4- Dioxo -3,4- Dihydropyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methoxy )( Phenoxy ) Phosphoryl ) Amine ) Isopropyl propionate (1c) At -5°C, tributylmagnesium chloride (0.57 mL, 0.98 mmol, 1.7 M THF) was added dropwise to a stirred solution of compound 1b (60 mg, 0.28 mmol) in DMPU (0.6 mL). The mixture was stirred at -5°C for 30 minutes and then at room temperature for 30 minutes. At -5°C, add a solution of ((perfluorophenoxy)(phenoxy)phosphoryl)-L-alanine isopropyl ester (0.25 g, 0.56 mmol) in anhydrous THF (2.5 mL) and The reaction mixture was stirred at room temperature for 8 hours. After the reaction was complete (TLC), water (15 mL) was added and the mixture was extracted with EtOAc (30 mL). Wash the organic phase with saturated sodium chloride solution (10 mL) and dry (Na₂ SO₄ ), filtered and concentrated, and the resulting crude product was purified by column chromatography on 230-400 silica gel using a 4-5% MeOH/DCM gradient to obtain the title compound (55 mg, 38%) as a solid. MS (ES+) [484.0]⁺ .¹ H NMR (DMSO-d₆ , 400 MHz) δ 1.15-1.20 (10H), 3.73-3.75 (1 H), 4.11-4.27 (4H), 4.84-4.90 (1H), 5.14 (1H), 5.51-5.53 (1H), 6.06-6.12 ( 1H), 6.26-6.27 (1H), 7.17-7.23 (3H), 7.36-7.40 (2H), 7.57-7.60 (1H), 11.37 (1H).Examples 2

(2S)-2-((((((2S,4S)-4-(4- Amine -2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methoxy )( Phenoxy ) Phosphoryl ) Amine ) Isopropyl propionate (2) According to the procedure described in step c of Example 1, trisatabin (TR-9) (50 mg, 0.23 mmol) was reacted with phosphorylating agent I-36 (0.26 g, 0.58 mmol) to obtain the title compound as a solid (30 mg, 26%). MS (ES+) 483.34 [M+H]⁺ .¹ H NMR (DMSO-d₆ , 400 MHz) δ 1.14-1.24 (9H), 3.32-3.38 (1H), 4.05-4.21 (4H), 4.84-4.26 (1H), 5.14 (1H), 5.68-5.70 (1H), 6.07-6.13 (1H ), 6.23-6.25 (1H), 7.16-7.24 (5H), 7.34-7.39 (2H), 7.59-7.61 (1H).Examples 3

(2S)-2-((((((2S,4S)-4-(4- Amine -2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methoxy )( Phenoxy ) Phosphoryl ) Amine ) Isopropyl propionate (3) According to the procedure described in Example 1, Step c, Trisatabin (50 mg, 0.23 mmol) was reacted with phosphorylating agent I-38 (0.24 g, 0.58 mmol) to give the title compound (40 mg, 35 %). MS (ES+) 481.0 [M-H]^- .¹ H NMR (DMSO-d₆ , 400 MHz) δ 1.14-1.20 (9H), 3.76-3.77 (1H), 4.10-4.18 (2H), 4.22-4.25 (2H), 4.84-4.87 (1H), 5.17-5.186 (1H), 5.69-5.70 (1H), 6.03-6.08 (1H), 6.24-6.26 (1H), 7.17-7.25 (5H), 7.36-7.40 (2H), 7.62-7.64 (1H).Examples 4

(2S)-2-((((((2S,4S)-4-(4- Amine -2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methoxy )( Phenoxy ) Phosphoryl ) Amine ) Isopropyl propionate (4) According to the procedure described in Example 1, Step c, Trisatabin (50 mg, 0.23 mmol) was reacted with phosphorylating agent I-37 (0.33 g, 0.58 mmol) to obtain the title compound (30 mg, 22 %). MS (APCI) 599.47 [M+H]⁺ . The compounds listed in Table 2 were prepared as pure diastereomers according to the procedure described in step c of Example 1, using suitable intermediates, I-numbered diastereomer-1 or I-numbered diastereomer Body-2. Table 2

Similarly, the compounds listed in Table 3 were prepared as pure diastereomers using the appropriate intermediates according to the procedure described in Example 1, step c. table 3

Record the NMR and MS data of all exemplified compounds to confirm their structure.Examples 35

(2S)-2-((((((2S,4S)-4-(2- Pendant -4- Palmitidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methoxy )( Phenoxy ) Phosphoryl ) Amine ) Isopropyl propionate (35 Diastereomer 1 and 35 Diastereomer -2) According to the method described in WO2008/030373, compounds 2 and 3 were acylated with palmitic anhydride to obtain the title compound.Examples 36

(2S)-2-((((((2S,4S)-4-(2- side Oxygen -4- Palmitidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methoxy )( Phenoxy ) Phosphoryl ) Amine ) Methyl propionate (36) According to the method described in WO2008/030373, compound 27 diastereomer-2 was acylated using palmitic anhydride to obtain the title compound.Comparative example

step a) (2S)-2-(( double (4- Methoxyphenyl )( Phenyl ) methyl ) Amine )-N-(2- Oxygen ion group -1,3,2- Oxaphos -2- base ) Acrylamide Under nitrogen, a solution of 2-chloro-1,3,2-oxathiophosane (0.542 g, 3.80 mmol) was added dropwise to (S)-2-((bis(4-methoxybenzene Group) (phenyl)methyl)amino)propanamide (1.40 g, 3.58 mmol) and triethylamine (0.60 ml, 4.30 mol) in an ice-cold solution in dichloromethane (8 ml). The reaction was maintained at room temperature and stirred over the weekend. The solution was cooled to 0°C and a solution of (third butylperoxy)trimethylsilane (1.16 g, 7.17 mmol) in heptane was slowly added. The reaction mixture was allowed to stir for 90 minutes, then concentrated in vacuo. The residue was suspended in ethyl acetate (10 mL), the hydrochloride salt was removed by filtration and the solvent was removed in vacuo. The residue was dissolved in anhydrous acetonitrile (10 mL) and the resulting solution was used in the next step without further purification. based on³¹ P-NMR determined the quantitative yield and 80% purity.step b) ((S)-2-(( double (4- Methoxyphenyl )( Phenyl ) methyl ) Amine ) Propionyl ) Amine Hydrogen Phosphate ((2S,4S)-4-(4- Amine -2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methyl ester Under nitrogen, DMAP (229 mg, 1.88 mmol) was added to a solution of compound Tr-9 (100 mg, 0.469 mmol) in anhydrous pyrimidine (5 mL), followed by slowly adding (2S)-2-(( Bis(4-methoxyphenyl)(phenyl)methyl)amino)-N-(2-oxo-1,3,2-oxathiolane)propionamide (361 mg, 0.563 mmol) in anhydrous acetonitrile (2 mL). Under nitrogen, the resulting solution was stirred at room temperature for 46 hours and then concentrated. The residue was purified by preparative HPLC on Gemini-NX 5m C18 (100×30 mm) using a gradient of 20% B to 80% B in 17 minutes at a flow rate of 35 mL/min. Solvent A: 95% water, 5% acetonitrile (10 mM ammonium acetate); solvent B: 10% water, 90% acetonitrile (10 mM ammonium acetate). The product-containing fractions were combined and lyophilized to obtain the title compound (80 mg, 26%). MS (ES+) 664.26 [M+H]⁺ .step c) ((S)-2- Aminopropionyl ) Amine Hydrogen Phosphate ((2S,4S)-4-(4- Amine -2- Pendant pyrimidine -1(2H)- base )-1,3- Dioxolane -2- base ) Methyl ester Water (50 mL) was added to the dichloromethane solution of the compound from the previous step (80.5 mg, 0.121 mmol), followed by acetic acid (500 mL). At room temperature, the solution was stirred for 12 minutes, then TFA (75 mL) was added and the resulting solution was stirred at room temperature for 5 minutes, diluted with toluene (10 mL), concentrated to dryness and dried under vacuum. The residue was dissolved in water (10 mL) containing 10% acetonitrile and washed with third butyl methyl ether (2×10 mL) containing 10% hexane. The aqueous layer was collected and freeze-dried overnight to obtain the desired product (80 mg) in the form of bis-TFA salt, which had a purity of about 75% according to LC-MS. The desired residue was further purified by preparative HPLC on Hypercarb (21.2×100 mm, I=271 nm) using a 0% to 35% acetonitrile/water gradient. The fractions containing the product were combined and lyophilized. MS (ES+) 364.10 [M+H]⁺ . use¹ H and¹³ C NMR confirmed the structure. NMR data of selected exemplary compounds:Chemical compound 8 Diastereomer -1 ¹ H NMR (DMSO-d₆ , 400 MHz) δ 0.81-0.84 (6H), 1.20-1.22 (11H), 1.59 (1H), 3.82-3.97(3H), 4.08-4.16 (2H), 4.22-4.23 (2H), 5.16 (1H), 5.67-5.69 (1H), 6.05-6.10 (1H), 6.23-6.24 (1H), 7.16-7.23 (m, 5H), 7.34-7.38 (m, 2H), 7.60-7.62 (m, 1H).Chemical compound 8 Diastereomer -2 ¹ H NMR (DMSO-d₆ , 400 MHz) δ0.81-0.84 (6H), 1.22-1.27 (11H), 1.57(1H), 3.81-3.89 (2H), 3.95-3.98 (1H), 4.05-4.07 (1H), 4.10-4.20 ( 3H), 5.128 (1H), 5.68-5.69 (1H), 6.13-6.14 (1H), 6.22-6.24 (1H), 7.16-7.21 (5H), 7.34-7.38 (2H), 7.58-7.60 (1H).Chemical compound 9 Diastereomer -1 ³¹ P NMR (DMSO-d₆ ) δ 4.354.¹ H NMR (DMSO-d₆ , 400 MHz) δ1.24-1.26 (3H), 3.98-4.01 (1H), 4.12-4.14 (2H), 4.27-4.29 (2H), 5.00-5.08 (2H), 5.16-5.18 (1H), 5.64- 5.66 (2H), 6.25-6.27 (1H), 6.34 (1H), 7.17-7.22 (2H), 7.31-7.33 (5H), 7.45-7.46 (2H), 7.55-7.59 (2H), 7.63-7.64 (1H ), 7.74-7.77 (1H), 7.95-7.97 (1H), 8.08-8.11 (1H).Chemical compound 9 Diastereomer -2 ³¹ P NMR (DMSO-d₆ ) δ 4.159.¹ H NMR (DMSO-d₆ , 400 MHz) δ1.25-1.26 (3H), 3.97-4.01 (1H), 4.08-4.16 (2H), 4.23-4.29 (2H), 5.04-5.16 (3H), 5.65-5.66 (1H), 6.26 ( 1H), 6.36-6.42 (1H), 7.17-7.24 (2H), 7.326 (5H), 7.41-7.49 (2H), 7.57-7.64 (3H), 7.74-7.76 (1H), 7.95-7.97 (1H), 8.10-8.12 (1H).Chemical compound 11- Diastereomer -1 ¹ H NMR (DMSO-d₆ , 400 MHz) δ0.23 (9H), 0.78-0.82 (3H), 1.08-1.12 (3H), 1.20-1.22 (3H), 1.44-1.49 (2H), 3.77-3.79 (1H), 4.09-4.23 ( 4H), 4.67-4.72 (1H), 5.16-5.16 (1H), 5.69-5.70 (1H), 6.04-6.10 (1H), 6.23-6.25 (1H), 7.15-7.24 (4H), 7.48-7.50 (2H ), 7.61-7.63 (1H).Chemical compound 11 Diastereomer -2 ¹ H NMR (DMSO-d₆ , 400 MHz) δ0.22-0.24 (9H), 0.78-0.82 (3H), 1.10-1.11 (3H), 1.22-1.24 (3H), 1.46-1.50 (2H), 4.05-4.07 (1H), 4.11- 4.22 (4H), 4.70-4.71 (1H), 5.14 (1H), 5.69-5.71 (1H), 6.07-6.11 (1H), 6.23-6.25 (1H), 7.16-7.24 (4H), 7.49-7.51 (2H ), 7.60-7.62 (1H). In order for the prodrug to target the liver, correct handling of the prodrug is crucial. The prodrug should be stable in the intestinal fluid and treated with liver enzymes in the first pass metabolism in the liver to form monophosphate. Subsequently, the monophosphate formed is anabolically metabolized in hepatocytes by cellular kinases to an active triphosphate substance. In addition, anticancer drugs should be toxic to proliferating cells. A suitable method for evaluating these properties of the compound is, for example, as described below.In the human intestine S9 section (HIS9) In the human liver S9 section (HLS9) Stability in Stock solutions (10 mM) of each test compound were prepared in DMSO and stored at -20°C. Before starting the experiment, the test compound was diluted to 500 µM in 50% acetonitrile in water. Prepare 50 mM potassium phosphate buffer (pH 7.4) in a total volume of 250 μL containing 5 mM MgCl₂ , 1 mM NADPH and 5 μM test compound reaction mixture. The reaction was initiated by adding the human liver or intestine S9 fraction so that the final concentration was 0.4 mg protein/mL (Xeno Tech). The reaction mixture was incubated on an orbital shaker at 37°C. At the desired time points (0, 10, 30 and 60 minutes), 50 μL aliquots were taken and the reaction was stopped by mixing with 150 μL of acetonitrile containing internal standard. From 500 µM solution by boiling the solution in human S9 (0.4 mg protein/mL), 5 mM MgCl₂ Prepare a standard solution of each test compound by diluting it to a final concentration of 5 µM in 50 mM potassium phosphate buffer (pH 7.4). The standards and samples were placed on ice for 30 minutes and then centrifuged at 3,000 g at 10°C for 20 minutes, and then 10 µL of supernatant was mixed with 200 µL of 50% acetonitrile in water. A 50% acetonitrile aqueous solution containing 0.5 µM of each test compound was injected into LC/MS-MS to measure product ions, declustering voltage (DP), collision energy (CE), and collision chamber outlet voltage (CXP) for LC/MS- MS method. The compound was separated using a C18 column with QTRAP5500 system. The mobile phase consists of solvent A (98% water, 2% acetonitrile, 0.1% acetic acid or 10 mM ammonium acetate) and solvent B (80% acetonitrile, 20% water, 0.1% acetic acid or 10 mM ammonium acetate). Compound dissolution was carried out with a solvent B gradient of 0% to 100%. Inject 5 µL standard points and samples for QTRAP5500 analysis. The amount of the parent compound was determined based on the peak area at each time point compared to the standard set at 5 µM. Using Excel software, determine the inherent clearance rate (CL_int ) And half-life (t_1/2 ).Cytotoxicity analysis 24 hours before compound addition, cells were seeded. Add each test compound (serial dilution from 100 μM) to Huh7 (1.5×10⁴ Cells/well) or HepG2 (1.5×10⁴ Cells/well) and cultured at 37°C for 5 days. The medium-only control was used to determine the minimum uptake value and the untreated cell value. At the end of the growth phase, XTT dye from Polysciences Europe GmbH was added to each well. The absorbance at 450 nm was read using Sunrise (Tecan) at a reference wavelength of 600 nM, and the medium-only control was used as a blank. Determine the 50% inhibition value (CC) by comparing the degree of inhibition plotted against the compound concentration (relative to the cell control)₅₀ ). The results of the dilution series were fitted to s-type dose-response curves. The compounds of the present invention were evaluated in these analyses to evaluate the stability in human intestine S9 (HIS9) and human liver S9 (HLS9), and cytotoxicity in HUH7, HEP3B, and HEPG2 cells. The results are summarized in Table B1. Table B1

na = not availableTriphosphate form analysis In the analysis, each compound was tested in triplicate. Fresh human-inoculated stem cells (Biopredic, France) were used in 12-well plates. Inoculate each well with 0.76×10⁶ Cells and 10 μM compound (0.1% DMSO) in DMSO in 1 mL of medium in CO₂ Incubate at 37°C for 8 hours. Inoculate Huh7 cells grown in DMEM with antibiotics and 10% fetal bovine serum in a 12-well plate, 2×10⁵ Cells/well. After 24 hours, add 1 mL of medium containing 10 µM compound and incubate the cells for another 6-8 hours. The culture was stopped by washing each well twice with 1 mL of ice-cold Hank's equilibrium solution pH 7.2, and then adding 0.5 mL of ice-cold 70% methanol. Immediately after adding methanol, the cell layer was detached from the bottom of the well with a cell scraper and sucked back and forth 5-6 times with an automatic pipette. The cell suspension was transferred to a glass bottle and stored at -20°C overnight. Subsequently, the samples were vortexed and centrifuged in an Eppendorf centrifuge 5417R at 14000 rpm for 10 minutes at 10°C. Each sample line consisted of various concentrations of prodrug, free nucleoside, and mono-, di-, and triphosphates. By inserting, transfer the supernatant to a 2 mL glass vial and perform a biological analysis according to the following: An internal standard (Indinavir) was added to each sample and the samples were analyzed on a two-column system coupled to a QTRAP 5000 mass spectrometer (10 µL injection volume). The two column system consists of two binary pumps, X and Y, two switching valves and an autosampler. The two HPLC columns used were Synergy POLAR-RP 50*4.6 mm, 4 µm particles and BioBasic AX 50*2.1 mm, 5 µm particles. The LC flow rate is 0.4-0.6 mL/min (a higher flow rate is used in the reconditioning step). The HPLC mobile phase for the POLAR-RP column is composed of 10 mmol/L ammonium acetate in 2% acetonitrile (mobile phase A) and 10 mmol/L ammonium acetate in 90% acetonitrile (mobile phase B). BioBasic AX column The HPLC mobile phase consists of 10 mmol/L ammonium acetate in 2% acetonitrile (mobile phase C) and 1% ammonium hydroxide in 2% acetonitrile (mobile phase D). The HPLC gradient of pump Y starts with 0% mobile phase B and is maintained for 2 minutes. During the loading phase, the mobile phase is passed through the POLAR-RP and BioBasic AX columns, and the prodrug, nucleoside, and internal standard are captured on the POLAR-RP column; and the nucleotides (mono-, di-, and triphosphates) are dissolved into BioBasic AX column and trapped here. In the next step, the flow from the POLAR-RP column is switched to MS and the mobile phase C is switched from pump X to BioBasic AX column. The compound on the POLAR-RP column was dissolved using a gradient of 0% B to 100% B in about two minutes and analyzed in a positive or negative mode using multiple reaction monitoring mode (MRM). In the final step, the flow from the BioBasic AX column was switched to MS and the phosphate was dissolved using a gradient of approximately 7 minutes up to a 50% D and analyzed in positive or negative mode using MRM. In the last step, adjust two more columns. Subsequently, the triphosphate concentration of each compound was determined by comparison with a standard curve prepared by analyzing standard samples with known concentrations of triphosphate. Run the standard on the same substrate as the test sample. Because of changes in phosphorylation levels between liver cell donors, internal standard compounds are required in each round of analysis to rank the results of different rounds with each other. In the specification and the scope of patent applications below, unless the context requires otherwise, the word "comprise" and its variations "comprises" and "comprising" should be understood to imply including the whole and steps shown , Whole group or step group but does not exclude any other whole, step, whole group or step group. The full text of all documents (including patents and patent applications) mentioned in this article is incorporated by reference.

Claims

A compound represented by formula Ia or a pharmaceutically acceptable salt thereof:

Where: R ¹ is NH ₂ ; R ² is F; R ¹³ is phenyl or naphthyl, wherein the phenyl or naphthyl is optionally substituted by R ²² ; R ¹⁵ is C ₁ -C ₃ alkyl; R ¹⁶ is H, C ₁ -C ₁₀ alkyl, C ₃ -C ₇ cycloalkyl, C ₃ -C ₇ cycloalkyl C ₁ -C ₃ alkyl, benzyl or phenyl; R ^{22 is} selected from halo and C ₃ -C ₆ cycloalkyl.

The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹⁵ is methyl.

The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹⁶ is C ₃ -C ₁₀ alkyl.

The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R ¹⁶ is 2-propylpentyl or 2-ethylbutyl.

The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹⁶ is benzyl.

The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹⁶ is C ₃ -C ₇ cycloalkyl.

A compound as claimed in claim 1 or a pharmaceutically acceptable salt thereof, wherein R ¹³ is phenyl, which is optionally substituted with R ²² .

The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein R ¹³ is phenyl.

If the compound of claim 1 or a pharmaceutically acceptable salt thereof is:

If the compound of claim 1 or a pharmaceutically acceptable salt thereof is:

A pharmaceutical composition comprising a therapeutically effective amount of a compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable adjuvant, diluent or carrier.

A use of the compound according to any one of claims 1 to 10 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for treating cancer.