Dioxolane analogues of uridine for the treatment of cancer
阅读说明：本技术 用于治疗癌症的尿苷的二氧戊环类似物 (Dioxolane analogues of uridine for the treatment of cancer ) 是由 R·贝特尔 A·埃内罗特 B·克拉松 F·欧伯格 于 2015-08-24 设计创作，主要内容包括：本发明提供了式(I)的化合物,其中,R<Sup>1</Sup>是OR<Sup>11</Sup>或NR<Sup>5</Sup>R<Sup>5</Sup>’；R<Sup>2</Sup>是H或F；R<Sup>5</Sup>是H、C<Sub>1</Sub>-C<Sub>6</Sub>烷基、OH、C(＝O)R<Sup>6</Sup>、O(C＝O)R<Sup>6</Sup>或O(C＝O)OR<Sup>6</Sup>；R<Sup>5</Sup>’是H或C<Sub>1</Sub>-C<Sub>6</Sub>烷基；R<Sup>6</Sup>是C<Sub>1</Sub>-C<Sub>6</Sub>烷基或C<Sub>3</Sub>-C<Sub>7</Sub>环烷基；R<Sup>13</Sup>是H、苯基、吡啶基、苄基、吲哚基或萘基,其中,所述苯基、吡啶基、苄基、吲哚基和萘基任选被1、2或3个R<Sup>22</Sup>取代；其它变量如权利要求所定义,所述化合物用于治疗癌症和相关的方面。<Image he="224" wi="700" file="DDA0002283077450000011.GIF" imgContent="drawing" imgFormat="GIF" orientation="portrait" inline="no"></Image>(The invention provides compounds of formula (I) wherein 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 、O(C＝O)R 6 OR O (C ═ O) OR 6 ；R 5 ' is H or C 1 ‑C 6 An alkyl group; r 6 Is C 1 ‑C 6 Alkyl or C 3 ‑C 7 A cycloalkyl group; r 13 Is H, phenyl, pyridyl, benzyl, indolyl or naphthyl, wherein said phenyl, pyridyl, benzyl, indolyl and naphthyl are optionally substituted by 1,2 or 3R 22 Substitution; the other variables are as defined in the claims, for use in the treatment of cancer and related aspects.)
1. A compound having the formula:
2. a pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable adjuvant, carrier or diluent.
3. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of cancer.
4. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of liver cancer.
5. Use of a compound according to claim 1, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament for the treatment of hepatocellular carcinoma.
Technical Field
The present invention relates to phosphorus prodrugs of troxacitabine and their derivatives for the treatment of cancer, in particular liver cancer such as hepatocellular carcinoma (HCC) and secondary liver cancer. The invention further relates to compositions and combinations comprising these compounds, and methods of their use in the treatment of cancer, particularly liver cancer such as HCC.
Background
Primary liver cancer is the sixth most common cancer in the world and is the second leading cause of cancer death. The most common liver cancer, which accounts for about 85% of all primary malignant liver cancers and continues to rise in incidence, is hepatocellular carcinoma (HCC), which is formed by hepatocytes that become malignant. Another type of cancer formed by hepatocytes is hepatoblastoma, a rare malignancy that occurs predominantly in children and accounts for approximately 1% of all childhood cancers and approximately 79% of all primary liver cancers under the age of 15. Secondary liver cancer or liver metastasis is cancer that initiates elsewhere in the body and subsequently spreads to the liver. Examples of secondary liver cancers include many common types of cancer, e.g., colon, rectal, lung, and breast cancer. Liver cancer can also be formed by other structures within the liver, such as bile ducts, blood vessels, and immune cells. Cholangiocarcinoma (cholangiocellular carcinoma and cholangiocellular cystadenocarcinoma) accounts for approximately 6% of primary liver cancers.
Although surgical resection and liver transplantation are potential treatments for early stage HCC, more than 20% of patients eventually relapse or encounter other problems, and most HCC diagnoses are performed at a too late stage of these treatments. Local treatments, such as radiofrequency ablation, have response rates in excess of 60%, but they are only suitable for a proportion of patients and are not always effective. The chemotherapy used to date has very limited effect on HCC, with response rates not exceeding 25% to date. Currently, sorafenib (sorafenib) is the only effective drug on the market to treat advanced or unresectable HCC, and therefore, other treatments for HCC are also highly desirable in order to reduce recurrence rates and improve overall survival rates.
Many nucleoside analogues have been found to have anti-cancer activity and they constitute a major class of chemotherapeutic agents widely used in the treatment of cancer patients. Such agents, known as antimetabolites, include various pyrimidine and purine nucleoside derivatives having cytotoxic activity.
Cellular nucleotide kinases phosphorylate nucleosides to their corresponding 5' -monophosphates and further to their diphosphates and subsequently to pharmacologically active triphosphates. Some nucleosides are known to be poorly active because they are not efficiently phosphorylated by kinases, or are not substrates for kinases at all. In the phosphorylation sequence, the first phosphorylation of a nucleoside analog is rate-limiting, while the second and third phosphorylation are less sensitive to nucleoside modification. Nucleoside monophosphates (nucleotides) themselves are generally unstable in blood and exhibit poor membrane permeability, and are therefore unsuitable for use as drugs. Nucleoside and nucleoside analogue triphosphates cannot be considered as potential drug candidates due to their high instability and poor cell permeability.
In particular, considerable activity has been observed against human cancer cell lines and xenografts of hepatocyte, prostate and kidney origin (cancer res, 55, 3008-.
In 2008, troxacitabine entered phase III clinical trials for acute myelogenous leukemia indications, but was not continuously registered. Phase II trials of termination of troxacitabine included breast, colorectal, pancreatic, melanoma, NSCLC, renal, prostate and ovarian tumors. Troxacitabine is typically administered by intravenous infusion, thereby exposing many tissues to the drug, regardless of the site of the cancer.
It has been shown that despite its hydrophilic nature, troxacitabine is transported into cells by passive diffusion, but that it accumulates only very slowly in cancer cells compared to other vector-transported nucleosides.
Derivatives of troxacitabine carrying a prodrug group on the cytosine base moiety are disclosed in WO2008/030373 and the relationship between the lipophilicity of the prodrugs and their antitumor activity is evaluated. This patent recognizes that base modification is desirable to avoid difficulties with esterases having 5' -OH modifications.
Phosphoramidate prodrugs on the 5' hydroxyl functionality of D-nucleosides have been successfully applied to antiviral drugs, for example sofosbuvir (sofosbuvir) for the treatment of HCV infection. Unmasking the sofosbuvir prodrug within the cell to expose the monophosphate is a complex, multi-step process involving several hydrolytic enzymes in a specific order.
The use of phosphoramidate prodrugs on cancer nucleosides has been less successful. Acelirin (Nuc-1031), a phosphoramidate prodrug of the D-nucleoside gemcitabine for the treatment of pancreatic cancer, is being developed by Nucana (see page 71 of WO2005012327 for structure). However, even though phosphoramidates are thought to enhance the lipophilicity and cellular permeability of the compounds, Acelarin prodrugs must also be administered in IV infusion form, thereby exposing many healthy tissues to cytotoxic metabolites.
Even less experience has been experienced with monophosphate prodrugs of L-nucleosides (e.g., troxacitabine). WO2008048128 discloses a few prodrugs of troxacitabine monophosphate, including the compound of example 14:
no cancer or other biological activity is disclosed for any of the compounds in the WO2008048128 specification or elsewhere in the academic literature. There is also no report of the entry of this prodrug into clinical trials. However, the inventors of WO2008048128 have disclosed prodrugs broadly similar to the D-nucleoside gemcitabine (Baraniak et al, Biorg MedChem 2014, 2133-. Kulic speculates that azidothymidine prodrugs tend to dephosphorylate nucleosides first, then phosphorylate into the active triphosphate species. Since the prodrug approach works with gemcitabine (which resembles RNA due to its substituted 2 'functional group) but not azidothymidine (which is 2' -deoxy and thus resembles DNA), WO2008048128 assumes that the prodrug of troxacitabine (which is a DNA analogue, albeit L-DNA) is likely to be inactive, resembling the azidothymidine prodrug.
Balzarini et al describe the HIV and HBV activity of CF1109 in Biochem Biophys Res Comm 225, 363-
Balzarini suggests that this phosphoramidate prodrug is about 250-fold less active against HIV than its parent nucleoside, 3TC, but this prodrug is "nearly as effective against HBV in Hep G2.2.15 cells". In other words, the addition of such a large methyl phosphoramidate prodrug group does not enhance antiviral efficacy in liver cell lines. Balzarini did not examine whether the prodrug was metabolized to 3TC prior to phosphorylation to the active triphosphate.
The present invention provides phosphorus prodrugs of troxacitabine (phosphorus produgs), especially liver-targeting prodrugs such as phosphoramidates, suitable for oral administration. These prodrugs have the advantage of improved cell permeability due to their increased lipophilicity compared to troxacitabine itself and due to the avoidance of the rate-limiting first phosphorylation step, the formation of active triphosphates can be more efficient. Further, the compounds of the present invention are metabolized primarily to active triphosphates in the liver, thereby providing high concentrations of active compounds in the target organ while maintaining minimization of side effects caused by toxicity in other organs.
Description of the invention
In one aspect, the present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt and/or solvate thereof:
wherein:
R1is OR11Or NR5R5’；
R2Is H or F;
R5is H, C1-C6Alkyl, OH, C (═ O) R6、O(C＝O)R6OR O (C ═ O) OR6；
R5'Is H or C1-C6An alkyl group;
R6is C1-C22Alkyl or C3-C7A cycloalkyl group;
R11is H or C1-C6An alkyl group;
R13is H, phenyl, pyridyl, benzyl, indolyl or naphthyl, wherein the phenyl, pyridyl, benzyl, indolyl isAryl and naphthyl optionally substituted by 1,2 or 3R22Substitution;
R15is H, C1-C6Alkyl radical, C3-C7Cycloalkyl radical, C3-C7Cycloalkyl radical C1-C3Alkyl, phenyl, benzyl or indolyl;
R15'is H or C1-C6An alkyl group; or
R15And R15'Together with the carbon atom to which they are attached form C3-C7A cycloalkylene group, wherein each C1-C6Alkyl is optionally selected from halogen, OR18And SR18Is substituted by each C3-C7Cycloalkyl radical, C3-C7Cycloalkylene, phenyl and benzyl optionally substituted by one or two independently selected from C1-C3Alkyl, halogen and OR18Substituted with a group of (1);
R16is H, C1-C10Alkyl radical, C2-C10Alkenyl radical, C3-C7Cycloalkyl radical, C3-C7Cycloalkyl radical C1-C3Alkyl, benzyl OR phenyl, any of which is optionally substituted by 1,2 OR 3 groups each independently selected from halogen, OR18And N (R)18)2Substituted with a group of (1);
each R18Independently is H, C1-C6Alkyl radical, C1-C6Haloalkyl or C3-C7A cycloalkyl group;
each R22Independently selected from halogen, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Haloalkyl, C1-C6Alkoxy radical, C1-C6Haloalkoxy, phenyl, hydroxy C1-C6Alkyl radical, C3-C6Cycloalkyl radical, C1-C6Alkylcarbonyl group, C3-C6Cycloalkyl carbonyl, carboxyl C1-C6Alkyl, hydroxy, amino CN and NO2Or to adjacent ring carbon atomsTwo R22The radicals may combine to form-O- (CR)23R23')1-6-O-；
R23And R23'Independently is H or C1-C3An alkyl group.
In one embodiment, the present invention provides a compound represented by formula I or a pharmaceutically acceptable salt and/or solvate thereof:
wherein:
R1is OR11Or NR5R5'；
R2Is H or F;
R5is H, C1-C6Alkyl, OH, C (═ O) R6、OC(＝O)R6OR OC (═ O) OR6；
R5'Is H or C1-C6An alkyl group;
R6is C1-C22Alkyl or C3-C7A cycloalkyl group;
R11is H or C1-C6An alkyl group;
R13is H, phenyl, pyridyl, benzyl, indolyl or naphthyl, wherein said phenyl, pyridyl, benzyl, indolyl and naphthyl are optionally substituted by 1,2 or 3R22Substitution;
R15is H, C1-C6Alkyl radical, C3-C7Cycloalkyl radical, C3-C7Cycloalkyl radical C1-C3Alkyl, phenyl, benzyl or indolyl;
R15'is H or C1-C6An alkyl group; or
R15And R15'Together with the carbon atom to which they are attached form C3-C7A cycloalkylene group, wherein each C1-C6Alkyl is optionally selected from halogen, OR18And SR18Of (2) aGroup substitution of each C3-C7Cycloalkyl radical, C3-C7Cycloalkylene, phenyl and benzyl optionally substituted by one or two independently selected from C1-C3Alkyl, halogen and OR18Substituted with a group of (1);
R16is H, C1-C10Alkyl radical, C2-C10Alkenyl radical, C3-C7Cycloalkyl radical, C3-C7Cycloalkyl radical C1-C3Alkyl, benzyl OR phenyl, any of which is optionally substituted by 1,2 OR 3 groups each independently selected from halogen, OR18And N (R)18)2Substituted with a group of (1);
each R18Independently is H, C1-C6Alkyl radical, C1-C6Haloalkyl or C3-C7A cycloalkyl group;
each R22Independently selected from halogen, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Haloalkyl, C1-C6Alkoxy radical, C1-C6Haloalkoxy, phenyl, hydroxy C1-C6Alkyl radical, C3-C6Cycloalkyl radical, C1-C6Alkylcarbonyl group, C3-C6Cycloalkyl carbonyl, carboxyl C1-C6Alkyl, hydroxy, amino CN, NO2And trimethylsilyl, or any two R attached to adjacent ring carbon atoms22The radicals may combine to form-O- (CR)23R23')1-6-O-；R23And R23'Independently is H or C1-C3An alkyl group.
The compounds of formula (I) may optionally be provided in the form of pharmaceutically acceptable salts and/or solvates. In one embodiment, there is provided a compound of the invention in the form of a pharmaceutically acceptable salt. In a second embodiment, there is provided a compound of the invention in the form of a pharmaceutically acceptable solvate. In a third embodiment, there is provided a compound of the invention in free form.
In bookIn a general embodiment of the invention, R1Is NR5R5'E.g. NH2Or NHC (═ O) C1-C6An alkyl group.
R2Typically H.
In a preferred embodiment, R1Is NH2，R2Is H.
In alternative embodiments, R1Is NH2，R2Is F.
Typically, in the compounds of formula (I), the moiety-NHC (R)15)(R15')-C(＝O)OR16Amino acid ester residues are formed, including natural and non-natural amino acid residues. Of particular interest are amino acid residues, wherein R15'Is hydrogen, R15Is methyl, isopropyl, isobutyl or benzyl. In the general configuration, R15'Is H, R15Is C1-C3Alkyl groups, for example, methyl, ethyl, propyl, isopropyl.
At R15'Is hydrogen, R15In compounds other than hydrogen, the configuration of the asymmetric carbon atom is typically that of an L-amino acid, thereby providing a compound having the stereochemistry shown in formula (Ia):
in a preferred configuration of the compounds of the formula Ia, R15Is methyl.
In a further configuration of the compound of formula Ia, R15Is benzyl.
In representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is H;
R13is phenyl, naphthyl or indolyl, any of which is optionally substituted by halogen such as bromo or C3-C4Cycloalkyl such as cyclopropyl;
R15is C1-C3An alkyl group;
R16is C1-C8An alkyl group.
In further representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is H;
R13is naphthyl;
R15is C1-C3An alkyl group;
R16is C1-C8Alkyl or benzyl.
In further representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is H;
R13is phenyl, optionally substituted in the 4-position by halogen, e.g. bromine or C3-C4Cycloalkyl such as cyclopropyl;
R15is methyl;
R16is C3-C8An alkyl group.
In further representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is H;
R13is phenyl;
R15is methyl;
R16is C3-C8An alkyl group.
In further representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is F;
R13is phenyl, naphthyl or indolyl, any of which is optionally substituted by halogen such as bromo or C3-C4Cycloalkyl such as cyclopropyl;
R15is C1-C3An alkyl group;
R16is C1-C8An alkyl group.
In further representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is F;
R13is naphthyl;
R15is C1-C3An alkyl group;
R16is C1-C8Alkyl or benzyl.
In further representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is F;
R13is phenyl, optionally substituted in the 4-position by halogen, e.g. bromine or C3-C4Cycloalkyl such as cyclopropyl;
R15is methyl;
R16is C3-C8An alkyl group.
In further representative configurations of the compounds of formula Ia,
R1is NH2；
R2Is F;
R13is phenyl;
R15is methyl;
R16is C3-C8An alkyl group.
In a further configuration, R15And R15' together with the carbon atom to which they are attached form C3-C7Cycloalkyl groups, for example, cyclopropyl or cyclobutyl.
R16Is usually C1-C10Alkyl or C3-C7A cycloalkyl group.
R16Includes C1-C3Alkyl groups, for example, methyl, ethyl, propyl, isopropyl. R16Preferred examples of (A) are methyl, R16Is isopropyl.
In one embodiment, R16Is C3-C10An alkyl group.
According to this embodiment, R16Representative examples of (A) include branched chain C5-C8An alkyl group. In one embodiment, R16At the branch point of (C)1A bit. In alternative embodiments, R16At the branch point of (C)2A bit. Generally, according to these embodiments, R15' is H, R15The stereochemistry of the attached carbon atom is that of an L-amino acid, thereby providing a compound of the general formula:
wherein R is161And R162Are identical or different C1-C3Alkyl radical, R163And R164Are identical or different C1-C3An alkyl group.
Typically, in the compound of formula (Ia'), R16Is 2-pentyl, i.e. R161Is propyl, R162Is methyl.
In a further typical configuration of the compound of formula (Ia'), R16Is 2-butyl, i.e. R161Is ethyl, R162Is methyl.
Typically, in the compound of formula (Ia'), R16Is 2-propylpentyl or 2-ethylbutyl, i.e. R163And R164Both are propyl or ethyl, respectively.
R16Further representative examples of (1) include C3-C7Cycloalkyl radicals, for example cyclohexyl.
R16A further representative example of (a) is cyclopentyl.
R16A further representative example of (a) is benzyl.
R13Typically phenyl, naphthyl or indolyl, any of which is optionally substituted by 1 or 2R22And (4) substitution.
In the inventionIn one embodiment, R13Is phenyl or naphthyl, any of which is optionally substituted.
In one embodiment of the invention, R13Is naphthyl.
In a preferred embodiment of the invention, R13Is phenyl.
R13Representative examples of (A) include optionally substituted with one, two or three R22A substituted phenyl group to provide a compound of formula (II-aa):
wherein when R is22When present, each R22Independently selected from halogen, C1-C6Alkyl radical, C2-C6Alkenyl and C1-C6An alkoxy group. Typically, the phenyl ring is unsubstituted or substituted by one R22And (4) substitution.
In one configuration of the compound of formula (II-aa), the phenyl ring is unsubstituted.
In a further configuration of the compound of formula (II-aa), the phenyl ring is substituted by one R22And (4) substitution. Typically, in this configuration, the substituent R22Is located at the 4-position of the benzene ring.
In one embodiment of the compounds of the invention, R13Is substituted in the 4-position by halogen (e.g. bromine) or C3-C4Cycloalkyl (e.g., cyclopropyl) substituted phenyl.
In one configuration of the compound of formula (II-aa), the phenyl ring is substituted by a carboxyl group C1-C6Alkyl substitution. One representative example of this configuration is that illustrated by the formula:
in a further configuration of the compound of formula (II-aa), the phenyl rings are flanked by two R on adjacent carbon atoms22Is substituted, and two R22Combined to form-O-CH2-O-, therebyThe structure forming the following parts:
R13further representative examples of (a) include optionally substituted pyridyl. Typically, the pyridyl moiety is unsubstituted or substituted with one or two substituents each independently selected from halogen, C1-C6Haloalkyl, C1-C6Alkyl radical, C2-C6Alkenyl radical, C1-C6Alkoxy, hydroxyl and amino.
In typical embodiments of the compounds of formula (I),
R1is NH2Or NHC (═ O) C1-C6An alkyl group;
R13is phenyl, naphthyl or indolyl, any of which is optionally substituted by halogen, C1-C3Alkyl radical, C1-C3Alkoxy radical, C3-C6Cycloalkyl or C1-C3Haloalkyl substitution;
R15'is H, R15Is C1-C3Alkyl or benzyl;
R16is C1-C10Alkyl or C3-C7A cycloalkyl group.
In typical embodiments of the compounds of formula (I) or (Ia),
R1is NH2Or NHC (═ O) C1-C6An alkyl group;
R13is phenyl or naphthyl, any of which is optionally substituted by halogen, C1-C3Alkyl radical, C1-C3Alkoxy radical, C3-C6Cycloalkyl or C1-C3Haloalkyl substitution;
R15'is H, R15Is C1-C3Alkyl or benzyl;
R16is C2-C10Alkyl or C3-C7A cycloalkyl group.
In further typical embodiments of the compounds of formula (I),
R1is NH2；
R2Is H;
R13is phenyl;
R15'is H, R15Is C1-C3An alkyl group;
R16is C1-C3Alkyl or cyclohexyl.
In further typical embodiments of the compounds of formula (I) or (Ia),
R1is NH2；
R2Is H;
R13is phenyl;
R15'is H, R15Is C1-C3Alkyl or benzyl;
R16is C3-C8Alkyl, cyclopentyl or cyclohexyl.
The compounds of the invention show anti-cancer activity, in particular against liver cancer such as HCC, and can be used as medicaments for the treatment of warm-blooded animals, especially humans, suffering from cancer. In particular, the compounds may be used as medicaments for the treatment of humans suffering from liver cancer, such as HCC.
To avoid undesirable side effects, especially toxicity in other organs, it is critical to deliver the drug to the tumor site while reducing exposure to normal tissues. The compounds of the present invention are stable in gastric fluid, but are readily metabolized by liver enzymes, so they can be absorbed in the stomach and transferred to the liver as masked cytotoxic agents, where they are absorbed, metabolized and formed into the active cytotoxic triphosphate form. Thus, the present invention provides compounds that are absorbed and processed primarily in the liver, thereby minimizing exposure to other organs in the body and toxic side effects.
Without being bound by theory, the anti-oncogenic activity of the compounds of the invention may act directly on the cellular processes of the rapidly acting tumorigenic cells of the cancer, but may additionally or alternatively exert their effect by modulating the microenvironment of the tumor, e.g., inhibiting angiogenesis, thereby starving the nourished tumor, resulting in inhibition of tumor growth.
The compounds of the invention are also useful in the treatment of secondary liver cancer, liver metastases, i.e. cancers originating from other organs in the body, such as the colon, lungs or breast, and metastasizing to the liver.
The invention also relates to a method of treating a warm-blooded animal, especially a human, having cancer, especially liver cancer such as HCC, comprising: administering an effective amount of a compound of formula (I) or any subgroup thereof.
The invention also relates to a method of treating a warm-blooded animal, especially a human, having secondary liver cancer, comprising: administering an effective amount of a compound of formula (I) or any subgroup thereof.
The use as a medicament or a method of treatment comprising systemically administering to a subject suffering from cancer an effective amount of a compound of formula (I).
In one aspect, the invention provides a pharmaceutical composition comprising a compound of formula (I) together with a pharmaceutically acceptable adjuvant, diluent, excipient or carrier.
In a further aspect, the present invention provides a pharmaceutical composition for the treatment of cancer comprising a compound of formula (I) together with a pharmaceutically acceptable adjuvant, diluent, excipient or carrier.
In a further aspect, the present invention provides a pharmaceutical composition for the treatment of liver cancer, such as HCC, comprising a compound of formula (I) together with a pharmaceutically acceptable adjuvant, diluent, excipient or carrier.
In a further aspect, the present invention provides a pharmaceutical composition for the treatment of secondary liver cancer comprising a compound of formula (I) together with a pharmaceutically acceptable adjuvant, diluent, excipient or carrier.
In a further aspect, the present invention relates to a method of preparing a pharmaceutical composition as recited herein, the method comprising: a pharmaceutically acceptable adjuvant, diluent, excipient and/or carrier is intimately mixed with a therapeutically effective amount of a compound of formula (I).
In a further aspect, the present invention provides a pharmaceutical composition for use in the above treatment or inhibition, said pharmaceutical composition further comprising one or more additional therapeutic agents.
The pharmaceutical compositions described above will generally comprise an effective amount (e.g., for use in humans) of a compound of formula (I), although sub-therapeutic doses of the compound of formula (I) may be employed 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 according to the individual requirements of each particular case. The reasons for affecting the dosage are, for example, the severity of the disease being treated, the age, weight, general health of the subject being treated, and the like. In the case of an anti-cancer effect, this effect may inhibit further tumor growth, reduce the likelihood of metastasis or eliminate metastasis, or cause tumor cell death, leading to tumor shrinkage, or prevent tumor regrowth after the patient's tumor has regressed.
In a further aspect, the present invention provides a compound of formula (I) for use as a medicament.
In a further aspect, the present invention provides a compound of formula (I) for use in the treatment of cancer.
In a further aspect, the present invention provides a compound of formula (I) for use in the treatment of liver cancer (e.g., HCC).
In a further aspect, the present invention provides a compound of formula (I) for use in the treatment of secondary liver cancer.
In a further aspect, the present invention provides a compound of formula (I) for use in the above treatment in combination with one or more additional cancer treatments, e.g. other anti-cancer drugs, surgery, immunotherapy and/or topical treatments, e.g. radiofrequency ablation.
In a further embodiment, the additional anti-cancer therapy is radiation therapy.
In one embodiment, the additional anti-cancer therapy is one or more other nucleoside analogs that exhibit potent 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, multidrug resistance reversing agents and biological response modifiers.
In one embodiment of this aspect, the additional therapeutic agent is a chemotherapeutic agent.
In a further aspect, the present invention provides a compound of formula (I) for use in the preparation of a medicament.
In a further aspect, the present invention provides a compound of formula (I) for use in the preparation of a medicament for the treatment of cancer.
In a further aspect, the present invention provides a compound of formula (I) for use in the preparation of a medicament for the treatment of liver cancer, e.g. HCC.
In a further aspect, the present invention provides a compound of formula (I) for use in the preparation of a medicament for the treatment of secondary liver cancer.
In a further aspect, the present invention provides a method for the treatment of cancer, said method comprising administering to a subject (e.g. a human in need thereof) a therapeutically effective amount of a compound of formula (I).
In a further aspect, the present invention provides a method for treating liver cancer (e.g., HCC), comprising administering to a subject (e.g., a human in need thereof) a therapeutically effective amount of a compound of formula (I).
In a further aspect, the present invention provides a method for treating secondary liver cancer, the method comprising: administering to a subject (e.g., a human in need thereof) a therapeutically effective amount of a compound of formula (I).
In a further aspect, the invention provides methods for the above treatment in combination with one or more additional cancer treatments, e.g., other anti-cancer drugs, surgery, immunotherapy and/or local treatments, e.g., radiofrequency ablation.
In one aspect, the present invention provides a method of treating primary or secondary liver cancer, said method comprising administering a pharmaceutical combination comprising a therapeutically effective amount of a compound of formula I, further comprising one or more additional therapeutic agents selected from the group consisting of chemotherapeutic agents, multidrug resistance reversing agents, and biological response modifiers.
In one embodiment of this aspect, the other therapeutic agent is a chemotherapeutic agent. In one aspect, the present invention provides a compound of formula (I) selected from the compounds described below, or a pharmaceutically acceptable salt thereof:
furthermore, the present invention relates to processes for the preparation of compounds of formula (I), to novel intermediates useful in the preparation of compounds of formula (I) and to processes for the preparation of such intermediates.
The terms "compound of formula (I)", "compound of the invention" or similar terms, as used hereinbefore and hereinafter, are intended 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 thereof.
The compounds of the present invention may be formulated into various pharmaceutical forms for administration purposes. As suitable compositions, all compositions commonly used for oral administration of drugs can be cited. To prepare the pharmaceutical compositions of this invention, an effective amount of the particular compound (optionally in addition salt form or solvate) as the active ingredient is intimately admixed with a pharmaceutically acceptable carrier, which may take a wide variety of forms depending on the form of preparation desired for administration. These pharmaceutical compositions are desirable in unit dosage forms suitable for oral administration. For example, in the preparation of compositions in oral dosage form, in the case of oral liquid preparations (e.g., suspensions, syrups, elixirs, emulsions and solutions), any conventional pharmaceutical media may be employed, such as, for example, water, glycols, oils, alcohols, and the like; or in the case of powders, pills, capsules and tablets, solid carriers may be employed, for example, starches, sugars, kaolin, lubricants, binders, disintegrating agents and the like. Because of their ease of administration, tablets and capsules represent the most advantageous oral dosage unit form in which case solid pharmaceutical carriers are obviously employed. Also included are solid form preparations which can be converted to liquid form preparations shortly before use.
It is particularly advantageous to formulate the above pharmaceutical compositions in unit dosage form for ease of administration and uniformity of dosage. Unit dosage form, as used herein, refers to physically discrete units suitable as unitary dosages, each unit containing a predetermined quantity of active ingredient suitable for the purpose of producing the desired therapeutic effect, in association with the required pharmaceutical carrier. Examples of such unit dosage forms are tablets (including scored or coated tablets), capsules, pills, powder packets, wafers, and the like, as well as divided portions thereof.
Generally, an effective daily dosage for anticancer will be considered to be from about 0.01 to about 700mg/kg, or from about 0.5 to about 400mg/kg, or from about 1 to about 250mg/kg, or from about 2 to about 200mg/kg, or from about 10 to about 150mg/kg of body weight. Suitably, the required dose is administered in two, three, four or more sub-dose forms at suitable time intervals per day. The sub-doses may be formulated in unit dosage forms, for example, each unit dosage form containing from about 1 to about 5000mg, or from about 50 to about 3000mg, or from about 100 to about 1000mg, or from about 200 to about 600mg, or from about 100 to about 400mg of the active ingredient.
The compound of the present invention can exhibit an anticancer effect alone and/or enhance the ability of another anticancer agent to exhibit an anticancer effect.
The compounds of the present invention may be represented as defined stereoisomers. The absolute configuration of such compounds may be determined using methods known in the art, e.g., X-ray diffraction or NMR measurements, and/or by the nature of the starting materials of known stereochemistry. Preferably, the pharmaceutical compositions according to the invention will comprise a preparation of the specified stereoisomer which is substantially stereoisomerically pure.
Pure stereoisomeric forms of the compounds and intermediates described herein are defined as: substantially free of other enantiomers or diastereoisomeric forms of the same basic molecular structure of said compound or intermediate. In particular, the term "stereoisomerically pure" relates to a compound or intermediate in at least 80% stereoisomeric excess (i.e., at least 90% of one isomer, and at most 10% of the other possible isomers) up to 100% stereoisomeric excess (i.e., 100% of one isomer, no other isomers), more particularly, 90% up to 100% stereoisomeric excess, even more particularly, 94% up to 100% stereoisomeric excess, most particularly, 97% up to 100% stereoisomeric excess. The terms "enantiomerically pure" and "diastereomerically pure" are to be understood in the same way, although the enantiomeric and diastereomeric excess of the mixture in question should be taken into account, respectively.
Pure stereoisomeric forms of the compounds and intermediates of the present invention may be obtained by using methods known in the art. Enantiomers can be separated from each other, for example, by selectively crystallizing their diastereomeric salts using optically active acids or bases. Examples thereof are tartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid and camphorsulfonic acid. Alternatively, enantiomers may be separated by chromatographic techniques using a chiral stationary phase. The pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction is stereospecific.
Preferably, if a particular stereoisomer is desired, the compound is synthesized by stereospecific methods of preparation. These processes will advantageously use enantiomerically pure starting materials.
Diastereomeric racemates of the compounds of the present invention may be obtained separately by conventional methods. Suitable physical separation methods which can advantageously be used are, for example, selective crystallization and chromatography, for example column chromatography.
When a phosphorus atom is present in the compounds of the present invention, the phosphorus atom may represent a chiral center. The chirality of this center is denoted as "R" or "S" according to Cahn-Ingold-Prelog precedence rules. When chirality is not indicated, it is considered to include both R-and S-isomers, as well as mixtures of the two, i.e., mixtures of diastereomers.
In a preferred embodiment of the present invention, stereoisomers are included in which the phosphorus atom has the S-configuration. These stereoisomers are denoted SP。
In other embodiments of the invention, stereoisomers are included in which the phosphorus atom has the R-configuration. These stereoisomers are represented by RP。
In other embodiments of the invention, mixtures of diastereomers are included, i.e., mixtures of compounds in which the phosphorus atom has the R-or S-configuration.
The invention also includes isotopically-labeled compounds of formula (I) wherein one or more atoms are replaced by an isotope of that atom, i.e., an atom having the same atomic number but an atomic weight different from the atomic weight usually found in nature. Examples of isotopes that can be incorporated into compounds of formula (I) include, but are not limited to: isotopes of hydrogen, for example,2h and3h (also denoted D and T for deuterium and tritium, respectively), isotopes of carbon, for example,11C、13c and14c, isotopes of nitrogen, for example,13n and15n, isotopes of oxygen, for example,15O、17o and18o, isotopes of phosphorus, for example,31p and32p, isotopes of sulfur, for example,35s, isotopes of fluorine, for example,18f, isotopes of chlorine, for example,36cl, isotopes of bromine, for example,75Br、76Br、77br and82br, and isotopes of iodine, for example,123I、124I、125i and131I. the choice of isotope included in the isotopically labeled compound will depend on the particular application for the compound. For example, for drug or substrate tissue distribution assays, the most commonly used compounds are those in which radioactivity is incorporatedAn isotope (for example,3h or14C) The compound of (1). For radiographic applications, such as Positron Emission Tomography (PET), positron emitting isotopes are used, for example,11C、18F、13n or15And O. The compounds of formula (I) incorporate heavy isotopes, for example, deuterium, i.e.,2h, may provide greater metabolic stability, e.g., may result in an increased in vivo half-life of the compound, or a reduced dosage requirement.
Isotopically-labelled compounds of the present invention can be prepared by employing procedures analogous to those described in the schemes and/or examples hereinbelow, using a suitable isotopically-labelled reagent or starting material in place of the corresponding non-isotopically-labelled reagent or starting material, or by conventional procedures known to those skilled in the art.
Pharmaceutically acceptable addition salts include the therapeutically active acid and base addition salt forms of the compounds of formula (I). Of interest are the free forms, i.e., the non-salt forms of the compounds of formula (I) or any subgroup thereof.
By treating the base form with such a suitable acid, pharmaceutically acceptable acid addition salts can be conveniently obtained. Suitable acids include, for example, inorganic acids, e.g., hydrohalic acids, e.g., hydrochloric or hydrobromic acids, sulfuric acid, nitric acid, phosphoric acid, and the like; or organic acids such as acetic, propionic, glycolic, lactic, pyruvic, oxalic (i.e., oxalic), malonic, succinic (i.e., succinic), maleic, fumaric, malic (i.e., malic), tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids. Conversely, the salt form may be converted to the free base form by treatment with a suitable base.
They may also be converted into their non-toxic metal or amine addition salt forms by treating the compounds of formula (I) containing the acidic proton with suitable organic and inorganic bases. Suitable base salt forms include, for example, ammonium salts, alkali and alkaline earth metal salts, e.g., lithium, sodium, potassium, magnesium, calcium salts, and the like, salts with organic bases, e.g., benzathine, N-methyl-D-glucamine, hydrabamine, and salts with amino acids, e.g., 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 can become stable in rings with aromatic character. Although not explicitly indicated in the structural formulae provided herein, such forms are included within the scope of the present invention.
The terms and expressions which have been employed in the specification abstract, specification, and claims are to be construed in accordance with the definitions set forth below, unless otherwise indicated. Each term is intended to be independent at each occurrence. These definitions apply regardless of whether a term is used alone or in combination with other terms, unless otherwise indicated. Terms or expressions used herein, which are not explicitly defined, should be interpreted as having the ordinary meaning as used in the art. To describe the same structure, chemical names, common names, and chemical structures may be used interchangeably. A structure is dominant if chemical compounds are referred to using both chemical structure and chemical name, and there is ambiguity between structure and name.
“Cm-CnAlkyl "by itself or in compound terms, e.g. Cm-CnHaloalkyl, Cm-CnAlkylcarbonyl group, Cm-CnAlkylamines, and the like, represent straight-chain or branched aliphatic hydrocarbon groups having the indicated number of carbon atoms, e.g. C1-C4Alkyl refers to alkyl groups having 1 to 4 carbon atoms. C1-C6Alkyl has the corresponding meaning and also includes all straight-chain and branched isomers of pentyl and hexyl. The preferred alkyl group for use in the present invention is C1-C6Alkyl, including methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl and n-hexyl, especially C1-C4Alkyl groups, for example, methyl, ethyl, n-propyl, isopropyl, tert-butyl, n-butyl and isobutyl. It is generally preferredMethyl and isopropyl. Alkyl may be unsubstituted alkyl or substituted with one or more substituents which may be the same or different, each substituent being independently selected from halogen, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH2NH (alkyl), -N (alkyl)2-NH (cycloalkyl), -O-C (═ O) -alkyl, -O-C (═ O) -aryl, -O-C (═ O) -cycloalkyl, -C (═ O) OH, and-C (═ O) O-alkyl. It is generally preferred that the alkyl group is an unsubstituted alkyl group unless otherwise specified.
“C2-CnAlkenyl "denotes a straight-chain or branched aliphatic hydrocarbon group containing at least one carbon-carbon double bond and having the number of carbon atoms specified, e.g. C2-C4Alkenyl means alkenyl having 2 to 4 carbon atoms; c2-C6Alkenyl means alkenyl having 2 to 6 carbon atoms. Non-limiting alkenyl groups include ethenyl, propenyl, n-butenyl, 3-methylbut-2-enyl, n-pentenyl and hexenyl. An alkenyl group may be unsubstituted alkenyl or substituted with one or more substituents which may be the same or different, each substituent being independently selected from: halogen, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH2NH (alkyl), -N (alkyl)2-NH (cycloalkyl), -O-C (═ O) -alkyl, -O-C (═ O) -aryl, -O-C (═ O) -cycloalkyl, -C (═ O) OH, and-C (═ O) O-alkyl. It is generally preferred that alkenyl groups are unsubstituted alkenyl groups, unless otherwise specified.
“C2-CnAlkynyl "denotes a straight or branched aliphatic hydrocarbon radical containing at least one carbon-carbon triple bond and having the indicated number of carbon atoms, e.g. C2-C4Alkynyl refers to alkynyl having 2 to 4 carbon atoms; c2-C6Alkynyl refers to alkynyl groups having 2 to 6 carbon atoms. Non-limiting alkenyl groups include ethynyl, propynyl, 2-butynyl, 3-methylbutynyl, pentynyl, and hexynyl. An alkynyl group can be unsubstituted or substituted with one or more substituents which can be the same or different, each substituent being independently selected from: halogen, halogen,Alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH2NH (alkyl), -N (alkyl)2-NH (cycloalkyl), -O-C (═ O) -alkyl, -O-C (═ O) -aryl, -O-C (═ O) -cycloalkyl, -C (O) OH and-C (O) O-alkyl. It is generally preferred that the alkynyl group is unsubstituted, unless otherwise specified.
The term "C" as used hereinm-CnHaloalkyl "denotes Cm-CnAlkyl in which at least one C atom is substituted by halogen (e.g. C)m-CnHaloalkyl groups may contain one to three halogen atoms), preferably chlorine or fluorine. Typical haloalkyl is C1-C2Haloalkyl, wherein halogen is suitably fluorine. Exemplary haloalkyl groups include fluoromethyl, difluoromethyl, and trifluoromethyl.
The term "C" as used hereinm-CnHydroxyalkyl "denotes Cm-CnAlkyl, wherein at least one C atom is substituted by one hydroxyl group. Typical Cm-CnHydroxyalkyl is C wherein one C atom is replaced by one hydroxy groupm-CnAn alkyl group. Exemplary hydroxyalkyl groups include hydroxymethyl and hydroxyethyl.
The term "C" as used hereinm-CnAminoalkyl "denotes Cm-CnAlkyl, wherein at least one C atom is substituted by one amino group. Typical Cm-CnAminoalkyl is C wherein one C atom is replaced by an amino groupm-CnAn alkyl group. Exemplary aminoalkyl groups include aminomethyl and aminoethyl.
The term "C" as used hereinm-CnAlkylene "represents a straight or branched chain divalent alkyl group having the specified number of carbon atoms. Preferred C for use in the present inventionm-CnAlkylene is C1-C3An alkylene group. Non-limiting examples of alkylene groups include: -CH2-、-CH2CH2-、-CH2CH2CH2-、-CH(CH3)CH2CH2-、-CH(CH3) -and-CH (CH)3)2)-。
The term "Me" refers to methyl and "MeO" refers to methoxy.
The term "Cm-CnAlkylcarbonyl group "represents formula Cm-Cnalkyl-C (═ O) -, where C ism-CnThe alkyl moiety is as described above. In general, "Cm-CnAlkylcarbonyl "is C1-C6alkyl-C (═ O) -. "Cm-CnAlkoxy "denotes the radical Cm-Cnalkyl-O-in which Cm-CnThe alkyl group is as described above. Of particular interest is C1-C4Alkoxy groups, including methoxy, ethoxy, n-propoxy, isopropoxy, tert-butoxy, n-butoxy and isobutoxy. Methoxy and isopropoxy are generally preferred. C1-C6Alkoxy has the corresponding meaning and includes all straight-chain and branched isomers of pentoxy and hexoxy.
The term "Cm-CnAlkoxycarbonyl is represented by the formula Cm-Cnalkoxy-C (═ O) -, where Cm-CnAlkoxy moieties are as described above. In general, "Cm-CnAlkoxycarbonyl "is C1-C6alkoxy-C (═ O) -.
The term "amino" denotes the group-NH2。
The term "halogen" denotes a halogen group, for example, fluorine, chlorine, bromine or iodine. Typically, the halogen group is fluorine or chlorine.
The term "aryl" refers to phenyl, biphenyl, or naphthyl.
The term "heterocycloalkyl" means 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 membered ring contains 1 heteroatom selected from O, S and N. In a second embodiment, the stable, saturated, monocyclic 3-7 membered ring contains 2 heteroatoms independently selected from O, S and N. In a third embodiment, the stable, saturated, monocyclic 3-7 membered ring contains 3 heteroatoms independently selected from O, S and N. ComprisesThe stable, saturated, monocyclic 3-7 membered ring of 1-3 heteroatoms independently selected from O, S and N can generally be a 5-7 membered ring, for example, a 5-or 6-membered ring. Heterocycloalkyl groups may be unsubstituted or substituted with one or more substituents which may be the same or different, each substituent being independently selected from: halogen, alkenyl, alkynyl, aryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH2NH (alkyl), -N (alkyl)2-NH (cycloalkyl), -O-C (═ O) -alkyl, -O-C (═ O) -aryl, -O-C (═ O) -cycloalkyl, -C (═ O) OH, and-C (═ O) O-alkyl. It is generally preferred that the heterocycloalkyl group is an unsubstituted heterocycloalkyl group unless otherwise specified.
The term "heteroaryl" denotes a stable mono-or bicyclic aromatic ring system containing 1 to 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 mono-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 mono-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 of the invention, the stable mono-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 of the invention, the stable mono-or bicyclic aromatic ring system contains four heteroatoms independently selected from O, S and N, each ring having 5 or 6 ring atoms.
One embodiment of heteroaryl includes flavone.
The term "C3-CnCycloalkyl "denotes a cyclic monovalent alkyl group having the indicated number of carbon atoms, e.g. C3-C7Cycloalkyl refers to a cyclic monovalent alkyl group having 3 to 7 carbon atoms. Preferred cycloalkyl groups for use in the present invention are C3-C4Alkyl groups, i.e., cyclopropyl and cyclobutyl. Cycloalkyl groups may be unsubstituted or substituted with one or more substituents which may be the same or different, each substituent being independently selected from: halogen, alkenyl, alkynylAryl, cycloalkyl, cyano, hydroxy, -O-alkyl, -O-aryl, -alkylene-O-alkyl, alkylthio, -NH2NH (alkyl), -N (alkyl)2-NH (cycloalkyl), -O-C (═ O) -alkyl, -O-C (═ O) -aryl, -O-C (═ O) -cycloalkyl, -C (═ O) OH, and-C (═ O) O-alkyl. It is generally preferred that the cycloalkyl group is unsubstituted, unless otherwise specified.
The term "amino Cm-CnAlkyl "represents the above C substituted by aminom-CnAlkyl, i.e. one hydrogen atom of the alkyl moiety is replaced by NH2-group substitution. In general, "amino group Cm-CnAlkyl "is amino C1-C6An alkyl group.
The term "amino Cm-CnAlkylcarbonyl "represents C abovem-CnAlkylcarbonyl wherein one hydrogen atom of the alkyl moiety is replaced by NH2-group substitution. In general, "amino group Cm-CnAlkylcarbonyl "is amino C1-C6An alkylcarbonyl group. Amino group Cm-CnExamples of alkylcarbonyl include, but are not limited to: glycyl ═ O) CH2NH2Alanyl: C (═ O) CH (NH)2)CH3Valyl: C ═ OCH (NH)2)CH(CH3)2Leucyl: C (═ O) CH (NH)2)(CH2)3CH3Isoleucyl: C (═ O) CH (NH)2)CH(CH3)(CH2CH3) And norleucyl: C (═ O) CH (NH)2)(CH2)3CH3And so on. This definition is not limited to naturally occurring amino acids.
The term "(═ O)" as used herein, when attached to a carbon atom, forms a carbonyl moiety. It should be noted that an atom may only carry one oxo group when the valency of the atom allows it.
The terms "monophosphate, diphosphate and triphosphate" refer to the group
As used herein, the radical position on any molecular moiety used in the definition may be at any position of that moiety, as long as it is chemically stable. When any variable occurs more than once in any part, each definition is independent.
The term "solvate" includes any pharmaceutically acceptable solvate that the compound of formula (I) and salts thereof is able to form. Such solvates are, for example, hydrates, alcoholates, e.g. ethanolates, propanolates, and the like, especially hydrates.
The term "prodrug" as used herein means a compound that is a precursor of a drug that is readily converted in vivo to the active compound by metabolic and/or chemical processes when administered to a subject.
The term "liver-targeting prodrug" as used herein means a prodrug that is metabolized to its active species primarily in the liver.
The term "liver cancer" as used herein is intended to include primary and secondary liver cancers, i.e., liver metastases which refer to cancers originating in the liver, as well as cancers of other organs, respectively.
The relevant terms are to be interpreted according to the definitions provided above and the usual usage in the technical field.
Generally, ChemDraw Ultra 12.0 is used to generate the name of the compound used in this application. In addition, a structure or portion of a structure is to be construed as including all stereoisomers of it if the stereochemistry of the structure or portion of the structure is not indicated, for example, by bold or dashed lines.
General synthetic methods
The compounds of the present invention may be prepared using a variety of methods, for example, those described in the illustrative synthetic schemes shown and described below. The starting materials and reagents used may be obtained from commercial suppliers or may be prepared according to literature procedures set forth in the literature using procedures well known to those skilled in the art.
Scheme 1 illustrates the general route for compounds of formula (I).
Reaction scheme 1
Commercially available derivatives of troxacitabine (1a) prepared as described above are condensed with the desired phosphoramidate reagent (1b) in an inert solvent such as an ether, e.g., diethyl ether or THF, or a halogenated hydrocarbon, e.g., dichloromethane, in the presence of a base, e.g., N-methylimidazole (NMI), or a grignard reagent, e.g., tert-butyl magnesium chloride, and the like, wherein Lg is a suitable leaving group, e.g., a halogen, e.g., chloride or an activated phenol, e.g., pentachlorophenol, p-nitrophenol, pentafluorophenol, and the like, to give the phosphoramidate derivative (1 c).
From phosphorus oxychloride (POCl)3) Initially, the phosphoramidate reagent (1b) used in the above-described scheme can be prepared using a two-step reaction in which Lg is chlorine, i.e., phosphoramidate chloride (phosphoroamiditate), as described in scheme 2.
Reaction scheme 2
In an inert solvent such as Et2In O, POCl3With the desired alcohol R13OH condensation to provide alkoxy or aryloxy dichlorophosphate (2 a). Followed by reaction with an amino acid derivative (2b) to provide a phosphoramidate chloride wherein R3Is H (2 c).
If desired, the phosphoramidate chloride (2c) obtained can be converted to have an activated phenol as leaving group (e.g., pentafluorophenol or p-NO)2Phenol), as generally described in scheme 3.
Reaction scheme 3
This conversion can be conveniently carried out by reacting the chloro derivative (2c) with the desired activated phenol in the presence of a base, for example triethylamine or a similar base, thereby providing the phosphorylating agents (3a) and (3 b).
The skilled person is aware of the individual Protecting Groups (PG) used in the above reaction schemes and describes their use and alternative protocols in detail in the literature, see, e.g., Greene t.w., Wuts p.g.m.protective groups in organic synthesis,2nd ed.new York, Wiley; 1995.
the term "N-protecting group" or "N-protected" as used herein refers to those groups which protect the N-terminus of an amino acid or peptide, or which prevent undesired reactions of the amino group during the course of synthesis, Greene discloses the commonly used N-protecting groups which include acyl groups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl, phthaloyl, o-nitrophenoxyacetyl, α -chlorobutyryl, benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl, and the like, sulfonyl groups such as benzenesulfonyl, p-toluenesulfonyl, and the like, carbamate-forming groups such as benzyloxycarbonyl, p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl, 3, 4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl, 2-nitro-4, 5-dimethoxybenzyloxycarbonyl, 3,4, 5-methoxybenzyloxycarbonyl, 3, 5-nitrobenzyloxycarbonyl, 2-isobenzoyl, 3, 5-benzyloxycarbonyl, 4-trimethyloxycarbonyl, 2-4, 5-benzyloxycarbonyl, 3, 4-benzyloxycarbonyl, 4, 5-trimethyloxycarbonyl, 4, isobenzoyloxy-benzoyloxy, isobenzoyl, 4, isobenzoyl, carbonoxycarbonyl, isobenzoyl, etc., and the like, including the like, preferably, carbonoxycarbonyl, isobenzoyl, carbonoxycarbonyl, isobenzoyl, and the like, and the.
In Greene above, hydroxyl and/or carboxyl protecting groups are also reviewed in detail, for example, methyl, substituted methyl ethers, for example, methoxymethyl, methylthiomethyl, benzyloxymethyl, t-butoxymethyl, 2-methoxyethoxymethyl, and the like, silyl ethers, for example, Trimethylsilyl (TMS), t-butyldimethylsilyl (TBDMS), tribenzylsilyl, triphenylsilyl, t-butyldiphenylsilyl, triisopropylsilyl, and the like, substituted ethyl ethers, for example, 1-ethoxymethyl, 1-methyl-1-methoxyethyl, t-butyl, allyl, benzyl, p-methoxybenzyl, benzhydryl, trityl, and the like, aralkyl groups, for example, trityl, and pixyl (9-hydroxy-9-phenylxanthene derivative, especially chloride). Ester hydroxy protecting groups include esters, for example, formates, benzylformates, chloroacetates, methoxyacetates, phenoxyacetates, pivalates, amantanates, trimethylbenzoates, benzoates, and the like. Carbonate hydroxy protecting groups include methyl, vinyl, allyl, cinnamyl, benzyl, and the like.
Detailed description of the embodiments
The following examples will now illustrate various embodiments and intermediates of the invention. These examples are intended to further illustrate the invention and are not intended to limit the scope of the invention. Compound names were generated using ChemDraw Ultra software, cambridge soft, version 12.0.2.
In addition to the above definitions, the following abbreviations are used in the synthetic schemes above and in the examples below. Abbreviations used herein have their accepted meaning if not defined.
Bn benzyl
BOP-Cl: bis (2-oxo-3-oxazolidinyl) phosphonyl chloride
DCC: dicyclohexylcarbodiimide
DCM: methylene dichloride
DIEA diisopropylethylamine
DMAP 4-dimethylaminopyridine
DMF/N, N-dimethylformamide
EtOAc: ethyl acetate
Et3N: triethylamine
EtOH ethanol
Et2Diethyl ether O
LC liquid chromatography
HOAc: acetic acid
HPLC and high performance liquid chromatography
MeCN: acetonitrile
MeOH: methanol
NT: 3-nitro-1, 2, 4-triazoles
on: overnight
Pg: protecting group
Ph to phenyl
rt: Room temperature
And (6) TEST: bis (triethoxysilyl) propyl-tetrasulfide
THF: tetrahydrofuran (THF)
TFA trifluoroacetic acid
TFAA trifluoroacetic anhydride
TIPS: triisopropylsilyl radical
Preparation of troxacitabine
Step 1): ((2, 2-Dimethoxyethoxy) methyl) benzene (Tr-1)
To a stirred solution of 2, 2-dimethoxyethanol (50g, 0.471mol) in DMF (200mL) at 0 deg.C was added benzyl bromide (56.03mL, 0.471mol) and NaOH (20.7g, 0.518mol), and the reaction mixture was stirred at room temperature for 16 h. After completion of the reaction (TLC), saturated aqueous sodium chloride (500mL) was added and the reaction mixture was extracted with DCM (1L) and dried (Na)2SO4) Concentrating the organic phase, purifying the crude product by silica gel column chromatography on 60-120 silica gelElution with 4-6% EtOAc/hexanes provided the title compound (60g, 60%) as a liquid.
Step 2): (5S) -5- ((4S) -2- ((benzyloxy) methyl) -1, 3-dioxolan-4-yl) -3, 4-dihydroxyfuran-2 (5H) -one (Tr-2)
L-ascorbic acid (44.9g, 0.255mol) was added to a solution of Compound Tr-1(60g, 0.306mol) in anhydrous acetonitrile (898mL), followed by pTSA monohydrate (15.5g, 0.076mol), and the reaction mixture was heated at 90 ℃ for 1 hour. After completion of the reaction (TLC), half the volume of acetonitrile was distilled off and the process was repeated twice. The solvent was completely removed to obtain a mixture of stereoisomers of the title compound (91 g). The product was used directly in the next step without further purification.
Step 3): (2R) -2- ((4S) -2- ((benzyloxy) methyl) -1, 3-dioxolan-4-yl) -2-hydroxyacetic acid (Tr-3)
At room temperature, compound Tr-2(91.7g, 0.297mol) was added to the stirred K2CO3(86.3g, 0.625mol) in water (509 mL). Slowly adding H2O2(80mL, 0.71mol, 30% v/v), and the solution was cooled to 0 ℃ followed by stirring for 24 hours. The solvent was removed under reduced pressure, EtOH (100mL) was added, and the mixture was heated at reflux for 30 minutes, then filtered. EtOH (100mL) was added to the resulting solid residue and the mixture was heated at reflux for 30 minutes (twice). The collected filtrate was concentrated in vacuo to give the title compound (90g) as a solid.
Step 4): (2S,4S) -2- ((benzyloxy) methyl) -1, 3-dioxolane-4-carboxylic acid (Tr-4a) and (2R,4S) -2- ((benzyloxy) methyl) -1, 3-dioxolane-4-carboxylic acid (Tr-4b)
Sodium hypochlorite (650mL, 0.881mol, 9-10% aqueous solution) was added dropwise to vigorously stirred compound Tr-3(90g, 0.294mol) and RuCl over a period of 30 minutes3.xH2O (1.22g, 0.0058mol) in an aqueous solution (ml, pH 8, room temperature). A1M NaOH solution was added to maintain the pH at a level of 8. The reaction mixture was stirred at room temperature for 3 hours and then heated at 35 ℃ for 12 hours. After completion of the reaction (TLC), 1.5N HCl was added to the reaction mixture at 0 deg.CUntil pH6 was reached, EtOAc (1L) was then added. The organic phase was washed with brine (2 × 100mL) and dried (Na)2SO4) Filtered and concentrated. The crude product obtained was purified by column chromatography on 230-400 silica gel eluting with 20% EtOAc/petroleum ether to give a mixture of isomers of compound 4a +4 b. The isomer was then separated by column chromatography over silica gel 230-400 using 0.9% MeOH/DCM and 0.1% AcOH as eluent to give the 2R isomer (20g, 28%).
Step 5): acetic acid (2S) -2- ((benzyloxy) methyl) -1, 3-dioxolan-4-yl ester (Tr-5)
To a solution of the compound Tr-4a (33g, 0138mol) in acetonitrile (660mL) were added pyridine (13.2mL) and lead acetate (79.8g, 0.180mol), and the mixture was stirred at room temperature for 16 hours. After completion of the reaction (TLC), the reaction mixture was filtered, the filtrate was concentrated, and the residue was added to EtOAc (500mL), washed with water (100mL) and saturated aqueous sodium chloride (100mL), and washed with Na2SO4And (5) drying. After removal of the solvent, the crude product was purified by column chromatography on 60-120 silica gel eluting with a 12-15% EtOAc/petroleum ether gradient to give the title compound (16g, 47%) as a liquid.
Step 6): acetic acid (2S) -2- (hydroxymethyl) -1, 3-dioxolan-4-yl ester (Tr-6)
To a stirred solution of compound Tr-5(16g) in dry methanol (160mL) was added Pd/C (3.2g, 20% w/w) and the reaction mixture was hydrogenated for 3 hours. After completion of the reaction (TLC), the reaction mixture was filtered through celite. The filtrate was concentrated under reduced pressure and the resulting crude title compound (10g, 97%) was used directly in the next step.
Step 7): acetic acid ((2S) -4-acetoxy-1, 3-dioxolan-2-yl) methyl ester (Tr-7)
To a stirred solution of compound Tr-6(5.74g, 0.0354mol) in pyridine (107mL) was added acetic anhydride (8.22mL, 0.080mol) at 0 deg.C, and the reaction mixture was stirred at room temperature for 16 hours. After completion of the reaction (TLC), the reaction mixture was quenched with dilute HCl (10mL) and extracted into EtOAc (100 mL). The organic phase was separated and dried (Na)2SO4) Filtered and concentrated. The crude product obtained is at 2Purification by column chromatography on 30-400 silica gel eluting with a gradient of 10-15% EtOAc/petroleum ether afforded the title compound (4.97g, 68%) as a liquid.
Step 8): acetic acid ((2S,4S) -4- (4- (benzylamino) -2-oxopyrimidin-1 (2H) -yl) -1, 3-dioxolan-2-yl) methyl ester (Tr-8a)
A mixture of N-benzoylcytosine (12.1g, 56.3mmol), ammonium sulfate (catalytic amount), and Hexamethyldisilazane (HMDS) (67.4mL, 418mmol) was refluxed for 1 hour. HMDS was removed under reduced pressure at 40 ℃ and the residue was added to anhydrous 1, 2-dichloroethane (57mL), a solution of compound Tr-7(5.7g, 27.9mmol) in anhydrous 1, 2-dichloroethane (57mL) was added, followed by the dropwise addition of TMSOTf (10.2mL, 45.7 mmol). The reaction mixture was stirred at room temperature for 1 hour, then NaHCO was added3Aqueous solution, and the mixture was stirred for 30 minutes. The resulting solid was filtered through celite, and the filtrate was added to EtOAc (200mL), washed with water (50mL), and dried (Na)2SO4). After removal of the solvent under reduced pressure, the crude product was purified by column chromatography on 230-400 silica gel eluting with a gradient of 10-15% EtOAc/petroleum ether to give a mixture of anomers which was further separated by SFC to give the title compound (3g, 30%) as a white solid.
Step 9): 4-amino-1- ((2S,4S) -2- (hydroxymethyl) -1, 3-dioxolan-4-yl) pyrimidin-2 (1H) -one (Tr-9)
A mixture of compound Tr-8a (3g) and saturated methanolic ammonia solution (180mL) was stirred at room temperature in a sealed tube for 16 hours. After completion of the reaction (TLC), the solvent was removed under reduced pressure and the crude product was purified by column chromatography on 230-400 silica gel eluting with a gradient of 10-13% MeOH/DCM to give the title compound (1.5g, 85%) as a solid.
1H NMR 400MHz DMSO-d6δ: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).
Preparation of 5-F-troxacitabine
Step 1): (2S,4R) -4- (4-benzoylamino-5-fluoro-2-oxopyrimidin-1 (2H) -yl) -1, 3-dioxolan-2-yl) methyl benzoate (5-F-Ta) and ((2S,4S) -4- (4-benzoylamino-5-fluoro-2-oxopyrimidin-1 (2H) -yl) -1, 3-dioxolan-2-yl) methyl benzoate (5-F-Tr-1b)
A mixture of 5-fluorobenzoylcytosine (9.1g, 39.5mmol), ammonium sulfate (catalytic amount) and hexamethyldisilazane (140mL) was refluxed for 14 hours. HMDS was removed under reduced pressure at 40 ℃ and the residue was added to anhydrous 1, 2-dichloroethane (50mL), a solution of the compound ((2S) -4-acetoxy-1, 3-dioxolan-2-yl) methyl benzoate (7g, 26.30mmol) in anhydrous 1, 2-dichloroethane (50mL) was added, followed by dropwise addition of TMS-OTf (11.6g, 52.6 mmol). The reaction mixture was stirred at room temperature for 2 hours, then NaHCO was added3An aqueous solution was added to the reaction mixture, and the mixture was stirred for another 30 minutes. The resulting solid was filtered off through celite, and the filtrate was added to EtOAc (500mL), washed with water (50mL), and dried (Na)2SO4). The solvent was removed under reduced pressure and the crude product was purified by column chromatography on 230-400 silica gel eluting with a gradient of 50-60% EtOAc/petroleum ether to give the pure title compound (1.7g, 18%) as a solid.
Step 2): 4-amino-5-fluoro-1- ((2S,4S) -2- (hydroxymethyl) -1, 3-dioxolan-4-yl) pyrimidin-2 (1H) -one (5-F-Tr)
A mixture of compound 5-F-Tr-1b (1.7g), saturated methanolic ammonia solution (34ml) was stirred in a sealed tube for 16 h, then the solvent was removed under reduced pressure and the crude product was purified by column chromatography on 230-400 silica gel eluting with a gradient of 5% MeOH/DCM to give the title compound (0.8g, 68%) as a solid.
The following phenols were prepared and used in the preparation of intermediates for the compounds of the present invention
Phenol 1
Step a):1- (3- ((tert-butyldimethylsilyl) oxy) phenyl) ethanone (Ph1-a)
Imidazole (4.46g, 65.5mmol) was added to a solution of 3-hydroxyacetophenone (4.46g, 32.8mmol) in DMF (6 mL). After 5 min, TBDMS-Cl (4.69g, 31.1mmol) in DMF (4mL) was added. The reaction mixture was stirred at room temperature for 90 minutes, then poured into hexanes containing 5% EtOAc (200mL) and washed with 1M HCl (60mL), water (60mL), saturated sodium bicarbonate (2 × 60mL), water (60mL) and brine (60 mL). With Na2SO4The organic layer was dried, filtered, concentrated, and the resulting residue was purified by flash chromatography on silica gel, eluting with hexanes/EtOAc, to give the title compound (5.7g, 69%).
Step b): tert-butyldimethyl (3- (prop-1-en-2-yl) phenoxy) silane (Ph1-b)
Methyl (triphenylphosphonium) bromide (10.2g, 28.4mmol) was suspended in anhydrous THF (30mL) under nitrogen and the suspension was cooled to 0 ℃. N-butyllithium (17.8mL, 28.4mmol) was added dropwise to the mixture, and the resulting solution was stirred at room temperature for 30 minutes. Ph1-a (5.7g, 22.8mmol) was added to the mixture and the reaction was allowed to proceed for 60 min at room temperature. The reaction was quenched with aqueous sodium bicarbonate and extracted with ether (50 mL). The organic layer was washed with sodium bicarbonate solution and dried (Na)2SO4) Filtered and concentrated. The resulting residue was purified by silica gel plug eluting with hexanes to give the title compound (3.9g, 69%).
Step c): tert-butyldimethyl (3- (1-methylcyclopropyl) phenoxy) silane (Ph1-c)
Diethyl zinc/hexane (439.2mmol) was added dropwise over a period of 10 minutes to a cooled (0 ℃ C.) solution of the alkene Ph1-b (3.9g, 15.7mmol) in 1, 2-dichloroethane (60mL) under a nitrogen atmosphere. Diiodomethane (6.32mL, 78.5mmol) was added dropwise and the resulting mixture was stirred at 0 ℃ for 30 minutes and then allowed to reach room temperature overnight. The mixture was poured into ice-cold ammonium chloride solution and extracted with ether. The organic layer was washed with saturated sodium bicarbonate and dried (Na)2SO4) Filtered and concentrated. The crude product was added to hexane and the residual di-product was discardedMethyl iodide. The hexane layer was concentrated and the crude product was used in the next step without further purification.
Step d): 3- (1-methylcyclopropyl) phenol (phenol 1)
Ph1-c (3.45g, 13.1mmol) was added to a 1M solution of tetrabutylammonium fluoride in THF (20mL, 20mmol) and the resulting solution was stirred at room temperature overnight. The reaction was quenched with 1M HCl (50ml) and extracted with ethyl acetate (100 ml). The organic layer was washed with brine (2 × 50ml) and dried (Na)2SO4) Filtered and concentrated. The residue was purified by flash chromatography on silica eluting with a mixture of 2-propanol, EtOAc and hexanes to give the title compound (0.56g, 29%). MS147.1[ M-H]-。
Phenol 2
The title compound was prepared from 4-hydroxyacetophenone (6.0g, 44.1mmol) using the method described for the preparation of phenol 1. The yield thereof was found to be 53%.
Phenol 3
Step a): 1- (3- (benzyloxy) phenyl) cyclopentanol (Ph3-a)
Iodine warmed with magnesium was added to a suspension of magnesium turnings (1.29 g, 52.8mmol) in anhydrous THF (50 ml). The mixture was refluxed and an approximately 5% solution of 3-bromophenol (13.9g, 52.8mmol) was added. When the reaction started, a solution of bromide was added dropwise, and then the mixture was refluxed for another hour. The mixture was cooled to about 5 ℃ and a solution of cyclopentanone (4.44g, 52.8mmol) in THF (50ml) was added dropwise. The mixture was stirred at room temperature for 72 hours, then the reaction was quenched with cold saturated ammonium chloride solution and extracted with diethyl ether (x 3). The organic phase was washed with brine and dried (Na)2SO4) Filtered and concentrated. The product was purified by silica gel chromatography (isohexane/EtOAc) to give the title compound (8.5g, 54)％)。
Step b): 1- (benzyloxy) -3- (cyclopent-1-en-1-yl) benzene (Ph3-b)
P-toluenesulfonic acid was added to a solution of Ph3-a (8.4g, 28.2mmol) in benzene (100 ml). The mixture was refluxed with a DMF trap for three hours, then cooled to room temperature, diluted with ether and washed with saturated sodium bicarbonate solution and brine. Drying (Na)2SO4) The organic phase was filtered and concentrated. The product was purified by silica gel chromatography (isohexane/EtOAc) to give the title compound (6.45g, 91%). MS249.4[ M-H ]]-。
Step c): 3-Cyclopentylphenol (phenol 3)
In Parr, a solution of Ph3-b (6.4g, 26mmol) in EtOAc (75ml) and EtOH (75ml) was hydrogenated at 22 ℃ and 40PSI overnight in the presence of 10% Pd on carbon (1.5 g). The catalyst was filtered off and washed with EtOAc and EtOH. The solvent was evaporated under reduced pressure and the product was chromatographed on silica gel (isohexane/EtOAc) to give the title compound (3.6g, 82%). MS 161.2[ M-H ]]-。
Phenol 4
Step a): tert-butyl (3-cyclopropylphenoxy) dimethylsilane (Ph4-a)
A mixture of (3-bromophenoxy) (tert-butyl) dimethylsilane (5.46g, 19mmol), cyclopropylboronic acid (2.12g, 24.7mmol), potassium orthophosphate (14.1g, 66.5mmol), tricyclohexylphosphine (0.53g, 1.9mmol) and Pd (OAc)2A suspension of (0.21g, 0.95mmol) toluene (80ml) and water (4ml) was stirred at 110 ℃ overnight. The slurry was diluted with ether and washed with water and brine. Drying (MgSO)4) The organic phase was filtered and concentrated. The crude product was purified by flash column chromatography (EtOAc/hexane) to afford the title compound (1.94g, 41%).
Step b): 3-Cyclopropylphenol (phenol 4)
1M tetrabutylammonium fluoride (10.1ml, 10.1mmol) was added to a solution of Ph4-a (1,94g, 7,81mmol) in THF (25 ml). The solution was stirred for 2 hours, then the solvent was evaporated, the residue was dissolved in EtOAc,with concentrated NH4Cl (aq) twice and once with brine. Drying (MgSO)4) The organic phase was filtered and concentrated. The crude product was purified by flash column chromatography (hexane/ethyl acetate, 9:1, containing 1% isopropanol) to afford the slightly impure title compound (1.24g, 119%).
Phenol 5
Step a): 2- (4-bromophenoxy) tetrahydro-2H-pyran (Ph5-a)
4-bromophenol (3.75g, 21.7mmol) was dissolved in 3, 4-dihydro-2H-pyran (16ml, 175mmol), catalytic amount of p-toluenesulfonic acid (15mg, 0.09mmol) was added, and the mixture was stirred at 22 ℃ for 45 minutes. The mixture was diluted with ether, washed with 1M NaOH (aq, x2), water, and dried (Na2SO4) Concentration gave the title compound (5.57g, 99%).
Step b): 2- (4-Cyclopropylphenoxy) tetrahydro-2H-pyran (Ph5-b)
A0.5M solution of cyclopropylmagnesium bromide in THF (6.5ml, 3.25mmol) was added over a 15 minute period to Ph5-a (552.5mg, 2,15mmol), ZnBr (144mg, 0.64mmol), tri-tert-butylphosphine tetrafluoroborate (35.6mg, 0.12mmol) and Pd (OAc)2(29.5mg, 0.13mmol) in THF (4 ml). The mixture was stirred at 22 ℃ for 90 minutes, then cooled on an ice bath and ice water (10ml) was added. The mixture was extracted three times with EtOAc and the extracts were washed with brine and subsequently dried (Na)2SO4) Filtering, and concentrating. The residue was purified by silica gel column chromatography (petroleum ether/EtOAc) to give the title compound (292mg, 62%).
Step c): 4-Cyclopropylphenol (phenol 5)
P-toluenesulfonic acid monohydrate (18.9mg, 0.1mmol) was added to a solution of Ph5-b (2.28g, 10.45mmol) in MeOH (15 ml). The mixture was heated at 120 ℃ for 5 min in a microwave reactor, then concentrated and purified by column chromatography on silica gel (petroleum ether/EtOAc). The resulting solid was crystallized from petroleum ether to give the title compound (1.08g, 77%).
Phenol 6
Step a): 1- (3-methoxyphenyl) cyclobutanol (Ph6-a)
A1M solution of 3-methoxyphenylmagnesium bromide in THF (2.11g, 99.8mmol) was added dropwise to a stirred solution of cyclobutanone (6.66g, 95mmol) in diethyl ether (65ml) at between 0 and 10 ℃. The mixture was stirred at 0-10 ℃ for three hours, and then, the mixture was added to ice-cold saturated NH4Cl (300ml) and water (300 ml). The mixture was stirred for 10 minutes and then extracted three times with ether. The organic phase was dried (Na)2SO4) Filtered and concentrated. The resulting crude product was purified by silica gel chromatography (isohexane/EtOAc) to give the title compound (16.9g, 86%).
Step b): 1-cyclobutyl-3-methoxybenzene (Ph6-b)
10% Pd/carbon (2.5g) was added to a solution of Ph6-a (15.4g, 86.1mmol) in ethanol (200ml), and the mixture was hydrogenated in Parr at 60 psi. After 18 hours, an additional 10% Pd on carbon (1.5g) was added and the mixture was further hydrogenated at 60psi for 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 chromatographed on silica gel (isohexane/EtOAc) to give the title compound (14.0g, 77%).
Step c): 3-Cyclobutylphenol (phenol 6)
A solution of 1M boron tribromide (18.1g, 72.2mmol) in DCM was added dropwise to a solution of Ph6-b (10.6g, 65.6mmol) in dry DCM (65ml) at 0 ℃. The mixture was stirred at-5 ℃ for 2.5 hours and then with cold saturated NH4The reaction was quenched with Cl solution and extracted three times with DCM. Drying (Na)2SO4) The organic phase was filtered and concentrated. The resulting crude product was purified by silica gel chromatography (isohexane/EtOAc) to give the title compound (9.73g, 88%).
Phenol 7
Step a): 1- (4- (benzyloxy) phenyl) cyclobutanol (Ph7-a)
A solution of 1- (benzyloxy) -4-bromobenzene (2.63g, 100mmol) in ether: THF (1:1, 100ml) was added dropwise to a suspension of magnesium turnings (2.43g) and traces of iodine in ether (50ml) over a period of ≈ 1 hour at reflux. When the addition was complete, the mixture was refluxed for four hours and then cooled to ≈ 0 ℃. Anhydrous THF (50ml) was added followed by slow addition of cyclobutanone (7.01g, 100mmol) in diethyl ether (50ml) and the mixture was allowed to reach room temperature. After stirring for two hours, cold saturated NH was added4Cl solution (500ml) and the mixture was stirred for 15 min and then extracted twice with EtOAc. The organic phase was washed with brine, dried over sodium sulfate and evaporated under reduced pressure. The product was purified by silica gel column chromatography to give the title compound (12.5g, 42%).
Step b): 4-Cyclobutylphenol (phenol 7)
Under an argon atmosphere, 10% Pd on carbon (2.55g, 21.5mmol) was added to a solution of Ph7-a (12.4g, 41.4mmol) in absolute EtOH (110ml) and the mixture was hydrogenated at 45psi at room temperature for 18 h. 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, concentrated, and the residue was crystallized from petroleum ether to give the title compound (3.15g, 51%).
Phenol 8
4- (1-methylcyclopentyl) phenol (phenol 8)
A solution of 1-methylcyclopentanol (2.00g, 20.0mmol) and phenol (2.07g, 22.0mmol) in pentane (50ml) was added dropwise to fresh AlCl over a period of 30 minutes3(1.33g, 10mmol) of pentane (100 ml). The resulting mixture was stirred at room temperature for 72 hours under nitrogen and then the reaction mixture was poured into water/ice and HCl (12M, 20mmol, 1.66 ml).The organic phase was washed with water (50ml) and brine (50ml) and dried (Na)2SO4) Filtered and concentrated. The crude product was purified by silica gel column chromatography (MeOH-DCM) to give the title compound (426mg, 12%).
Phenol 9
Step a): 2- (4-bromo-3-methylphenoxy) tetrahydro-2H-pyran (Ph9-a)
pTs (16mg, 0.086mmol) was added to a solution of 4-bromo-3-cresol (4.0g, 21.4mmol) in 3, 4-dihydro-2-H-pyran (16ml, 175 mmol). The reaction mixture was stirred at room temperature for 1 hour, then diluted with ether and washed with 1M NaOH (aq) and water. Drying (Na)2SO4) The organic phase was filtered and concentrated. The crude product was purified by silica gel column chromatography (EtOAc/heptane) to give the title compound (3.32g, 57%).
Step b): 2- (4-cyclopropyl-3-methylphenoxy) tetrahydro-2H-pyran (Ph9-b)
Ph9-a (3.12g, 11.5mmol), ZnBr2(2.59g, 11.5mmol), Tri-tert-butylphosphine tetrafluoroborate (0.2g, 0.69mmol), and Pd (OAc)2(258mg, 1.15mmol) was placed in the flask, and the flask was purged with nitrogen several times. THF (10ml) was added with stirring, followed by dropwise addition of 0.5M cyclopropylmagnesium bromide/THF (35ml, 17.4mmol) over a period of 5 minutes. The mixture was stirred at room temperature, then filtered through a plug of celite, eluting with MeOH. The solution was concentrated and the crude product was purified by column chromatography over silica gel (EtOAc/heptane) to give the title compound (1.69g, 57%).
Step c): 4-cyclopropyl-3-methylphenol (phenol 9)
Ph9-b (1.70g, 7.30mmol) was dissolved in MeOH (20ml) and pTsxH was added2O (318mg, 1.67 mmol). The mixture was stirred at 22 ℃ for 30 minutes and then concentrated. The crude product was purified by column chromatography (EtOAc/heptane) to give the title compound (704mg, 65%).
Phenol 10
Step a): 4-cyclopropyl-1-methoxy-2-methylbenzene (Ph10-a)
4-bromo-1-methoxy-2-methylbenzene (4.39g, 21.9mmol) was reacted with cyclopropylmagnesium bromide as described in step b of Ph9 to give the title compound (1.54g, 43%).
Step b): 4-cyclopropyl-2-methylphenol (phenol 10)
Under nitrogen atmosphere at 0 deg.C, BBr3(5ml, 5mmol) was added to a solution of Ph10-a (1.54g, 9.49mmol) in DCM (7.5 ml). The reaction was stirred for 2 hours, then quenched with MeOH (3ml) and concentrated. The crude product was dissolved in EtOAc and washed with brine. Drying (Na)2SO4) The organic phase was filtered and concentrated. The crude product was purified by silica gel column chromatography to give the title compound (826mg, 59%). MS 147.11[ M-H ]]-。
Phenol 11
4-cyclopropyl-3-methoxyphenol (phenol 11)
The title compound was prepared from 4-bromo-3-methoxyphenol (1.11g, 5.49mmol) according to the method described for the preparation of phenol 9. The yield is 40%.
Phenol 12
Step a): 3- (dimethylamino) -1- (3-hydroxyphenyl) propan-1-one (Ph12-a)
A few drops of HCl were added to a solution of 3-acetylphenol (4.08g, 30mmol), paraformaldehyde (4.05g, 45mmol) and dimethylamine hydrochloride (2.69g, 33mmol) in absolute EtOH (100ml) and the reaction mixture was refluxed for 18 h. Dimethylamine hydrochloride (0.55eq, 1.22g), paraformaldehyde (0.5eq, 1.35g) and HCl (0.5ml) were further added, and the reaction mixture was refluxed for another 4 hours and then cooled to room temperature. The precipitated white solid was collected, washed with cold EtOH (50mL) and cold acetone (10mL), followed by lyophilization to give the title compound (2.59g, 38%) which was used directly in the next step without further purification.
Step b): cyclopropyl (3-hydroxyphenyl) methanone (phenol 12)
NaH (60% dispersion in mineral oil) (1.13g, 28.2mmol) was added portionwise to a stirred suspension of trimethyl sulphoxide iodide (6.20g, 28.2mmol) in DMSO (100ml) at room temperature. After 1h, solid Ph12-a (2.59g, 11.3mmol) was added portionwise with stirring and cooling. The reaction mixture was stirred at room temperature for 40 hours, then poured into cold water (200ml) and extracted with DCM (3 × 100 ml). With saturated NH4The organic phase was washed with aqueous Cl (2X100mL) and dried (Na)2SO4) Filtered and concentrated. The resulting crude product was purified by silica gel column chromatography (MeOH/DCM) to give the title compound (883mg, 48%).
Phenol 13
Step a): cyclopropyl (4-hydroxyphenyl) methanone (Ph13)
The p-hydroxy-gamma-chlorobenzone ketone (4.95g) was added in portions to a NaOH solution (8ml, aq, 50% w/w) followed by NaOH (35ml, aq, 25% w/w) over a period of about 30 minutes, followed by a batch addition of p-hydroxy-gamma-chlorobenzone ketone (4.95 g). The temperature was lowered to 140 ℃ and NaOH (8g) was added. After 90 minutes, water (10ml) was added and after a further 60 minutes the reaction mixture was cooled, diluted with water and neutralized to pH7 with HOAc (. apprxeq.27-30 ml). The precipitate formed was filtered, washed with water and dried in vacuo. The solid was in CHCl at 40 deg.C3(200ml) for 10 minutes and then at room temperature overnight. The slurry was heated to 40 ℃ over a period of 30 minutes and then filtered. Drying (MgSO)4) The filtrate was filtered and concentrated to ≈ 70 ml. Hexane was added to form an oil, which eventually became crystalline. The slurry was filtered and washed with CHCl3The solid was washed with hexane and dried,the title compound (4.15g, 51%) was obtained.
Phenol 14
Step a): 3- (1-hydroxy-2, 2-dimethylpropyl) phenol (Ph14-a)
t.Bu-MgBr (1.5eq) was added dropwise over a period of 30 minutes to a cold (-10 ℃ C.) mixture of 3-hydroxybenzaldehyde (2.00g, 16.4mmol) in diethyl ether (20 ml). During the addition, THF (20ml) was added. The mixture was brought to 23 ℃ and stirred for 6 hours. More t.Bu-MgBr (0.7eq) was added and the mixture was stirred overnight, then cooled and saturated NH added4The reaction was quenched with aqueous Cl. EtOAc was added to the mixture followed by 1M aqueous HCl until a homogeneous mixture was obtained. The phases were separated, the organic phase was washed with brine and dried (Na)2SO4) Filtered and concentrated. The resulting crude product was purified by column chromatography to give the title compound (1.1g, 37%).
Step b): 1- (3-hydroxyphenyl) -2, 2-dimethylpropan-1-one (Ph14)
Adding into a dried round flask
Phenol 15
1- (4-hydroxyphenyl) -2, 2-dimethylpropan-1-one (Ph15)
4-hydroxybenzaldehyde (3g, 24.6mmol) was reacted as described in the preparation of phenol 14 to give the title compound (538mg, 17%).
Amino acid 1
Step a): (S) - (S) -2- ((tert-butyloxycarbonyl) amino) propionic acid sec-butyl ester (AA1-a)
L-Boc-alanine (2.18g, 11.5mmol) was dissolved in anhydrous DCM (40ml) and the alcohol (R) -butan-2-ol (938mg, 12.6mmol) was added. The mixture was cooled to about 5 ℃ and EDC (3.31g, 17.2mmol) was added in portions followed by DMAP (140mg, 1.15 mmol). The mixture was allowed to reach room temperature and stirred overnight, then diluted with ethyl acetate (-300 ml) and the organic phase was washed three times with saturated sodium bicarbonate solution and once with brine. The organic phase was dried over sodium sulfate and concentrated under reduced pressure. Chromatography on silica gel eluting with isohexane and 10% ethyl acetate afforded the title compound (2.78g, 98%).
Step b): (S) - (S) -sec-butyl 2-aminopropionate (AA1-b)
A mixture of AA1-a (2.77g, 11.3mmol) and p-toluenesulfonic acid monohydrate (2.15g, 11.3mmol) in EtOAc (45ml) was stirred at 65 ℃ for 16 h and then concentrated under reduced pressure. The resulting residue was crystallized from ether to give the title compound (3.20g, 89%).
Amino acid 2
(S) - (R) -2-aminopropionic acid (pent-2-yl) ester (AA2)
Following the procedure described for the preparation of AA1, but using (R) -pentan-2-ol instead of (R) -butan-2-ol, the title compound was obtained (4.6 g).
Amino acid 3
(S) - (S) -2-aminopropionic acid (pent-2-yl) ester (AA3)
The title compound (8.3g) was obtained according to the method described for the preparation of AA1, using (S) -pentan-2-ol instead of (R) -butan-2-ol.
The following intermediates were prepared and may be used in the preparation of the compounds of the present invention
Intermediate 1
Step a): (R) -4-Fluorobenzyl 2- ((tert-Butoxycarbonyl) amino) propionate (I-1a)
Boc-L-AlaOH (19.92mmol), DMAP (1.99mmol) and (4-fluorophenyl) methanol (23.9mmol) were dissolved in CH2Cl2(100 ml). To this solution triethylamine (23.9mmol) was added followed by EDCl (23.9mmol) and the resulting reaction mixture was stirred at room temperature under nitrogen overnight. The reaction mixture is treated with CH2Cl2(100ml) diluted with saturated NaHCO3Washed with aqueous solution (2 × 50ml) and saturated aqueous NaCl solution (2 × 50ml), dried (Na)2SO4) And concentrated. The resulting residue was purified by silica gel column chromatography eluting with n-hexane-EtOAc (95:5 to 60:40) to give the title compound (4.44g) as a white waxy solid. MS:296[ M-H ]]-。
Step b): (R) -2-Aminopropionic acid 4-fluorobenzyl ester (I-1b)
Compound I-1a (14.93mmol) was dissolved in 4M HCl/dioxane (40ml), stirred at room temperature for 30 minutes and evaporated to dryness to give the hydrochloride salt of the title compound (3.4g) as a white powder. MS 198[ M + H ]]+。
Step c): 4-Fluorobenzyl (2R) -2- ((chloro (phenoxy) phosphoryl) amino) propionate (I-1)
PhOPOCl is added at-78 deg.C2(4.28mmol) was added dropwise to CH of Compound I-5b (4.28mmol)2Cl2To the solution, triethylamine (8.56mmol) was then added dropwise. The resulting reaction mixture was stirred at-78 ℃ under Ar and brought to room temperatureOvernight. The reaction mixture was evaporated on silica gel and purified by chromatography (n-hexane/EtOAc (88:12) - (0: 100)). The title compound (769mg) was obtained.31P-NMR(CDCI3) Delta 7.85(s) and 7.54(s) (R)PAnd SPDiastereoisomers).
Intermediate 2
Step a): (S) - (R) -2- ((tert-Butoxycarbonyl) amino) propionic acid sec-butyl ester (I-2a)
L-Boc-alanine (2.18g, 11.5mmol) was dissolved in anhydrous DCM (40ml) and the alcohol (R) -butan-2-ol (938mg, 12.6mmol) was added. The mixture was cooled to about 5 ℃ and EDC (3.31g, 17.2mmol) was added in portions followed by DMAP (140mg, 1.15 mmol). The mixture was allowed to reach room temperature and stirred overnight, then diluted with ethyl acetate (-300 ml) and the organic phase was washed three times with saturated sodium bicarbonate solution and once with brine. The organic phase was dried over sodium sulfate and concentrated under reduced pressure. Chromatography on silica gel eluting with isohexane and 10% ethyl acetate afforded the title compound (2.78g, 98%).
Step b): (S) - (R) -2-Aminopropionic acid sec-butyl ester (I-2b)
A mixture of I-10a (2.77g, 11.3mmol) and p-toluenesulfonic acid monohydrate (2.15g, 11.3mmol) in EtOAc (45ml) was stirred at 65 ℃ for 16 h and then concentrated under reduced pressure. The resulting residue was crystallized from ether to give the title compound (3.20g, 89%).
Step c): (2S) - (R) -2- (((4-Nitrophenoxy) (phenoxy) phosphoryl) amino) propionic acid sec-butyl ester (I-2)
Phenyl dichlorophosphate (1eq) was added to a solution of compound I-10b (3.15g, 9.92mmol) in DCM (75ml) at-30 deg.C under a nitrogen atmosphere, followed by dropwise addition of triethylamine (2 eq). The mixture was allowed to reach room temperature and stirred overnight, then cooled to about 5 ℃, solid 4-nitrophenol (1eq, 15mmol) was added followed by dropwise addition of triethylamine (1eq, 15mmol), and the mixture was stirred at room temperature for 4 hours, then concentrated under reduced pressure, diluted with ethyl acetate (40ml) and ether (40ml) 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 eluting with isohexane-ethyl acetate to give the title compound (4.19g, 79%).
The following compounds were prepared according to the method described for the preparation of I-2, using the appropriate alcohol
Intermediate 6, diastereomer-1&-2
The two diastereomers of compound I-6 were separated by SFC to give I-6-dia-1 and I-6-dia-2.
Intermediate 7
Step a): (S) -2-Aminopropionic acid cyclooctyl ester (I-7a)
To a slurry of L-alanine (1.7g, 19.1mmol) and cyclooctanol (25ml, 191mmol) in toluene (100ml) was added p-toluenesulfonic acid monohydrate (3.6g, 19.1 mmol). The reaction mixture was heated at reflux temperature for 25 hours and water was removed from the reaction using a Dean-Stark trap. The mixture was concentrated under reduced pressure and the residue was kept under vacuum overnight. To the residue (27g) was added diethyl ether (100 ml). The white precipitate was collected by filtration, washed with diethyl ether (3 × 50ml) and dried in vacuo to give the title compound (4.84g, 68%).
Step b): (2S) -Cyclooctyl 2- (((4-Nitrophenoxy) (phenoxy) phosphoryl) amino) propanoate (I-7) Compound I-7a was reacted as described in the preparation of step c of I-2 to give the title compound (4.7g, 76%)
Intermediate 8
Cycloheptyl (2S) -2- (((4-nitrophenoxy) (phenoxy) phosphoryl) amino) propionate (I-22)
Following the procedure described for the preparation of compound I-7, but using cycloheptanol (27ml, 224mmol) instead of cyclooctanol, the title compound was obtained (5.72g, 55%).
Intermediate 9
Cyclohexyl (2S) -2- (((4-nitrophenoxy) (phenoxy) phosphoryl) amino) propanoate (I-23)
Following the procedure described for the preparation of step c of I-2, but using cyclohexyl (S) -2-aminopropionate instead of 3, 3-dimethylbutyl (S) -2-aminopropionate, the title compound was obtained (10.6g, 82%).
Intermediate 10
(S) -2- ((bis (4-nitrophenoxy) phosphoryl) amino) propionic acid 2-ethylbutyl ester (I-10)
2-Ethyl butyl (S) -2-aminopropionate (5g, 14.49mmol) was added to a solution of bis (4-nitrophenyl) chlorophosphate (6.14g, 17.1mmol) in DCM (50ml), the mixture was cooled in an ice bath and Et was added dropwise3N (4.77ml, 34.2 mmol). After 15 min, the cooling was removed and the reaction mixture was stirred at 23 ℃ until the reaction was completed according to TLC. Then, ether was added, the mixture was filtered, and the filtrate was concentrated and purified by silica gel column chromatography to give the title compound (2.05g, 82%).
Intermediate 11
Step a): (S) -2-Aminopropanoic acid isopropyl ester (I-11a)
At 0 deg.C, adding SOCl2(29ml, 400mmol) was added dropwise to L-propyl acetateHCl salt of amino acid (17.8g, 200mmol) in isopropanol (700 ml). The suspension was stirred at room temperature overnight and then concentrated to give the title compound (29.2g, 87%).
Step b): (2S) -2- ((((((S) -1-isopropoxy-1-oxoprop-2-yl) amino) (4-nitrophenoxy) phosphoryl) -amino) propanoic acid isopropyl ester (I-11)
A solution of 4-nitrophenyl dichlorophosphate (1.8g, 7mmol) in DCM was added dropwise to a solution of the amine I-11a (2.35g, 14mmol) and triethylamine (7.7ml, 56mmol) in DCM at-60 ℃. The reaction mixture was allowed to reach room temperature, stirred overnight, concentrated, 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 eluting with isohexane-ethyl acetate to give the title compound (1.6g, 50%).
Intermediate 12
Step a): neopentyl (S) -2- ((tert-butoxycarbonyl) amino) propionate (I-12a)
EDAC and DMAP were added portionwise to a solution of Boc-alanine (18.9g, 100mmol) and neopentyl alcohol (13.0ml, 120mmol) in DCM (200ml) at-5 ℃. The reaction mixture was allowed to reach room temperature and stirred for 72 hours. EtOAc (700mL) was added and the organic phase was washed with saturated NaHCO3The solution was washed three times, once with brine and then concentrated. The resulting residue was purified by column chromatography, eluting with hexane-EtOAc (90/10 to 80/20), to give the title compound (21g, 81%).
Step b): (S) -2-aminopropionic acid neopentyl ester (I-12b)
P-toluenesulfonic acid (15.6g, 82.0mmol) was added to a solution of Boc protected amine I-12a (21.1g, 82.0mmol) in EtOAc (330mL) at-65 ℃. The reaction mixture was stirred at-65 ℃ for 8 hours and then allowed to reach room temperature overnight. The mixture was then filtered and concentrated to give the title compound (21g, 78%).
Neopentyl (2S) -2- ((((((S) -1- (neopentyloxy) -1-oxoprop-2-yl) amino) (4-nitrophenoxy) -phosphoryl) amino) propionate (I-12)
4-Nitrophenol dichlorophosphate was added dropwise to a solution of amine I-12b (3.90g, 24.5mmol) in DCM (100ml) over a period of 1 hour at-50 ℃. The reaction mixture was allowed to reach room temperature, stirred overnight, concentrated, then diluted with ether and kept at room temperature overnight. The mixture was filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel chromatography, eluting with isohexane-ethyl acetate, to give the title compound (4.8g, 77%).
Intermediate 32
(2S) - (R) -2- (((Perfluorophenoxy) (phenoxy) phosphoryl) amino) propionic acid sec-butyl ester (I-32)
Et was added at-70 ℃ under nitrogen over a period of 15 minutes3N (10.9ml, 78.1mmol) was added dropwise to a stirred solution of the pTs salt of sec-butyl (S) - (R) -2-aminopropionate (12.0g, 37.7mmol) in DCM (50 ml). To the mixture was added a solution of phenyl dichlorophosphate (5.61ml, 37.7mmol) in DCM (50ml) over a period of 1 hour. The reaction mixture was stirred at-70 ℃ for a further 30 minutes, then warmed to 0 ℃ over a period of 2 hours and stirred for 1 hour. Pentafluorophenol (6.94g, 37.7mmol) and Et were added over a period of 20 minutes3A solution of N (5.73ml, 41.1mmol) in DCM (30ml) was added to the mixture. The crude mixture was stirred at 0 ℃ for 18 hours and then concentrated. The residue was added to THF (100ml), and the insoluble material was filtered off and washed several times with THF. The solvent was evaporated and the residue was triturated with methyl tert-butyl ether. The insoluble material was filtered off and washed with methyl tert-butyl ether. The combined filtrates were concentrated and the crude solid was sonicated with n-hexane/EtOAc (80: 20; 100 ml). The solid was filtered off and washed with n-hexane/EtOAc (80:20) to give the pure phosphorus stereoisomer of the title compound as a white solid (2.3g, 13%).
Intermediate 33
(2S) -ethyl 2- (((perfluorophenoxy) (phenoxy) phosphoryl) amino) propionate (I-33)
The pure phosphorus stereoisomer of the title compound was prepared according to the procedure described for I-32, but starting from the HCl salt of ethyl (S) -2-aminopropionate (11.0g, 71.1 mmol). Yield: 8.56g, 27%.
Intermediate 34
(2S) -2- (((Perfluorophenoxy) (phenoxy) phosphoryl) amino) propanoic acid 2-ethylbutyl ester (I-34)
The pure phosphorus stereoisomer of the title compound was prepared according to the procedure described for I-32, but starting from the pTs salt of 2-ethylbutyl (S) -2-aminopropionate (18.8g, 54.4 mmol). Yield: 27.0g, 99%.
LC-MS 496.44[M+H]+。
Intermediate 35
(2S) -butyl 2- (((Perfluorophenoxy) (phenoxy) phosphoryl) amino) propionate (I-35)
Phenyl dichlorophosphate (12.4ml, 83.1mmol) was added to a cold (-20 ℃ C.) slurry of butyl (S) -2-aminopropionate (26.4g, 83.1mmol) in dichloromethane (200 ml). The mixture was stirred for 10 minutes and then Et was added dropwise over 15 minutes3N (25.5ml, 183 mmol). The mixture was stirred at-20 ℃ for 1 hour and then at 0 ℃ for 30 minutes. The mixture was kept cold in an ice bath, and the perfluorophenol (15.3g, 0.08mol) was added followed by dropwise addition of Et3N (11.6ml, 0.08 mol). The mixture was stirred overnight and slowly brought to 20 ℃. Adding diethyl ether, filtering the mixture through celite, concentrating, purifying by silica gel column chromatography, and purifying with stoneOil ether/EtOAc (9:1->8:2) elution. The appropriate fractions were combined, concentrated, and crystallized from petroleum ether-EtOAc (9:1) to give the pure phosphorus stereoisomer of the title compound as a white solid (2.23g, 5.8%).
Intermediate 36
Step a): l-alanine isopropyl ester hydrochloride (I-36a)
Thionyl chloride (80.2g, 0.674mol,1.5eq) was added to 2-propanol (400ml) over a period of 30 minutes at-7 to 0 ℃ with cooling, followed by addition of L-alanine (40.0g, 0.449mol) at 0 ℃. A flow indicator and a scrubber with a mixture of 27.65% sodium hydroxide (228g) and water (225g) were connected to the outlet. The reaction mixture was stirred at 67 ℃ for two hours, then at 70 ℃ for one hour, and at 20-25 ℃ overnight. The reaction mixture was distilled at 47-50 ℃ under reduced pressure (250-50mBar) using a 60 ℃ bath. When the distillation became very slow, toluene (100ml) was added to the residual oil and the distillation was continued at 48-51 ℃ under reduced pressure (150-50mBar) using a 60 ℃ bath until becoming very slow, methyl tert-butyl ether (tBME) (400ml) was added to the residual oil and the two-phase system was allowed to crystallize at 34-35 ℃ with efficient stirring. When crystallization was observed, the mixture was cooled to 23 ℃ over a period of one hour, and the precipitate was isolated by filtration. The filter cake was washed with tBME (100ml) and dried under reduced pressure to constant weight without heating to give the title compound (67.7g, 90%) as a white solid.
Step b): (S) -isopropyl 2- (((S) - (perfluorophenoxy) (phenoxy) phosphoryl) amino) propionate (I-36)
Phenyl dichlorophosphate (62.88g, 0.298mol, 1.0eq) was added to a solution of L-alanine isopropyl ester hydrochloride (50.0g, 0.298mol) in DCM (310ml) at 0 ℃ under a nitrogen atmosphere, and the addition was completed by washing with DCM (39 ml). The mixture was cooled and triethylamine (63.35g, 0.626mol, 2.1eq) was added over a period of 70 minutes with cooling, maintaining the temperature no higher than-14 ℃ and washing with DCM (39ml) to complete the addition. The mixture was stirred for one hour at-15 to-20 ℃ and then heated to-8 ℃ and a solution of pentafluorophenol (60.38g, 0.328mol, 1.1eq) and triethylamine (33.19g, 0.328mol, 1.1eq) in DCM (78ml) was added over a period of 42 minutes with cooling, maintaining the temperature no higher than 0 ℃ and washing with DCM (39ml) to complete the addition. The mixture was stirred at 0 ℃ for one hour, followed by stirring at +5 ℃ overnight. The precipitate formed was removed by filtration and the filter cake was washed with DCM (95 ml). The combined filtrates were washed with water (2 × 190ml) at 5 ℃. The organic phase was distilled at 32-38 ℃ under reduced pressure (650-600mBar) and distillation was continued until a residual volume of about 170ml was obtained, obtaining a partially crystalline material. Ethyl acetate (385ml) was added and the resulting clear solution was distilled at 43-45 ℃ under reduced pressure (300-250 mBar). Distillation was continued until a residual volume of about 345ml was obtained. The clear solution was cooled to 36 ℃ and seeded with (S) -isopropyl 2- (((S) - (perfluorophenoxy) (phenoxy) phosphoryl) amino) propionate (20mg) (prepared as described in j.org.chem., 2011, 76, 8311-asant 8319) to induce crystallization. The mixture was cooled to 27 ℃ over a period of one hour, then n-heptane (770ml) was added over a period of 47 minutes and the mixture was stirred for an additional 37 minutes. Triethylamine (6.03g, 0.2eq) was added and the mixture was stirred at 23-25 ℃ overnight. The precipitate was separated by filtration. The filter cake was washed with ethyl acetate: n-heptane (1:9, 80ml) and dried under reduced pressure (less than 0.1mBar) without heating to constant weight to give the title compound (75.64g, 56%) as a white crystalline material.
1H NMR(CDCl3,300MHz)δ7.38-7.32(m,2H),7.27-7.24(m,2H),7.23-7.19(m,1H),5.10-4.98(m,1H),4.20-4.08(m,1H),4.03-3.96(m,1H),1.46(dd,7.2,0.6Hz,3H),1.26-1.23(2xd,6H)；
13CNMR(CDCl3,100MHz)δ172.7(d,J＝8.8Hz),150.4(d,J＝7.1Hz),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＝0.82Hz),125.8(d,J＝1.4Hz),120.3(d,J＝5.0Hz),69.8,50.6,(d,J＝1.9Hz),21.8(d,J＝1.9Hz),21.2(d,J＝4.4Hz)；
The crystalline properties and NMR spectroscopic data of the title compound were consistent with published data (j. org. chem., 2011, 76, 8311-8319), thus demonstrating the S stereochemistry of the phosphorus atom of the title compound.
Intermediate 37
Step a): (S) -2-Aminopropionic acid cyclohexyl ester (I-37a)
Acetyl chloride (4.2ml, 59.3mmol) was added dropwise to a stirred solution of cyclohexanol (50ml), followed by the addition of L-phenylalanine (4.0g, 24.2 mmol). The reaction mixture was heated to 100 ℃ for 16 h, then concentrated under reduced pressure, triturated with ether/hexane (1:1), dried to give the title compound (6g, 88%) as a white solid, which was used directly in the next step without further purification.
Step b): (S) -cyclohexyl 2- (((S) - (perfluorophenoxy) (phenoxy) phosphoryl) amino) propionate (I-37)
To a stirred solution of compound I-37a (7.0g, 24.6mmol) in anhydrous DCM (42ml) was added triethylamine (7.17ml, 51.5mmol) dropwise over 30 minutes at-70 deg.C followed by addition of a solution of phenyl dichlorophosphate (5.15g, 34.5mmol) in anhydrous DCM (21ml) over 1 hour. The reaction mixture was stirred at-70 ℃ for a further 30 minutes, then warmed to 0 ℃ over 2 hours and stirred for 1 hour. To the mixture was added a solution of pentafluorophenol (4.94g, 26.8mmol) and triethylamine (3.74ml, 26.8mmol) in anhydrous DCM (28ml) over 1 hour. The mixture was stirred at 0 ℃ for 4 hours and then held at 5 ℃ for 16 hours. The reaction mixture was filtered, and the filtrate was concentrated under reduced pressure. The crude solid was dissolved in EtOAc (300ml), washed with water (50ml), dried and the solvent removed under reduced pressure. The solid obtained was triturated with 20% EtOAc/hexanes, filtered, washed with hexanes, and dried to give the single diastereomer of the title compound (3.0g, 21%) as a solid.
Intermediate body 38
(2S) -isopropyl 2- (((4-nitrophenoxy) (phenoxy) phosphoryl) amino) propionate (I-38)
To a stirred solution of 4-nitrophenyl dichlorophosphate (5g, 19.8mmol) in anhydrous DCM (40ml) was added a solution of phenol (1.86g, 19.8mmol) and triethylamine (3ml, 21.8mmol) in anhydrous DCM (50ml) at-78 deg.C over a period of 30 minutes. The mixture was stirred at this temperature for 60 minutes and then transferred to another flask containing a solution of compound (S) -isopropyl 2-amino propionate (3.3g, 19.8mmol) in dry DCM (40ml) at-5 ℃ over a period of 15 minutes. To this mixture was added a second crop of TEA (6ml, 43.3mmol) over a 20 minute period at-5 ℃. The mixture was stirred at 0 ℃ for 3 hours, and then the solvent was removed under reduced pressure. The residue was added to EtOAc (200mL), washed with water (50mL), and Na2SO4Drying and removal of the solvent under reduced pressure gave a crude oil which was purified by column chromatography using a 0-20% EtOAc/hexane gradient and 230-400 mesh silica gel to give a mixture of diastereomers in a ratio of about 1: 1. The two diastereomers were separated by SFC to give the title compound as a solid, isomer 1(1.5g, 20%) and isomer 2(1.5g, 18%).
The compounds listed in Table 1 were prepared and the diastereomers isolated according to the procedure described for the preparation of intermediate I-38, using the appropriate amino acid ester and phenol.
TABLE 1
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