WO2020073942A1 - Tlr7 agonist prodrug, preparation method therefor and medical use thereof - Google Patents

Abstract

The present invention relates to a TLR7 agonist prodrug, a preparation method therefor and the medical use thereof. In particular, the present invention relates to a TLR7 agonist prodrug as shown in formula (I), a preparation method therefor, a pharmaceutical composition containing the derivative, and the use thereof as a therapeutic agent, in particular as a TLR7 agonist, wherein each substituent in the general formula (I) is the same as that defined in the description.

Description

TLR7 agonist prodrug, preparation method thereof and application in medicine Technical field

The present disclosure relates to a new TLR7 agonist prodrug represented by the general formula (I), a preparation method thereof, a pharmaceutical composition containing the prodrug, and its use as a therapeutic agent, especially as a TLR7 agonist.

Background technique

Toll-like receptors (TLRs) are a class of important protein molecules involved in innate immunity. TLRs are monomeric transmembrane non-catalytic receptors, usually expressed in sentinel cells such as macrophages and dendritic cells, and can recognize structurally conserved molecules produced by microorganisms. Once these microbes break through physical barriers such as the skin or intestinal mucosa, they are recognized by TLRs, which in turn activate immune cell responses. (Mahla, RS. Et al., Front Immunol. 4: 248 (2013)). The immune system's ability to recognize pathogenic microorganisms is due in part to the widespread existence of Toll-like immune receptors.

There are at least 10 different TLRs in mammals. Some of these receptor ligands and corresponding signal cascade amplification have been identified. TLR7 is a member of the subgroup of TLRs (TLRs 3, 7, 8, and 9) and is limited to the endosomal compartment of cells that specifically detect non-self nucleic acids. TLR7 can induce the secretion of type I interferon after activation, and plays an important role in innate immunity and acquired immunity.

A large number of preclinical animal experiments have shown that TLR7 agonists can effectively inhibit the growth of cancer cells, and even completely eradicate tumors in animal tumor models (Sidky et al., Cancer Res, 1992, 52 (13), 3528-33; Dumitru et al., Cancer Immunol. Immunother. 2009, 58, 575-587). Clinically, imiquimod, a TLR7 agonist for the treatment of basal cell carcinoma, has been approved for marketing.

The patent applications related to TLR7 agonists that have been published so far are WO2005025583, WO2007093901, WO2008011406, WO2009091032, WO2010077613, WO2010133882, WO2011031965 and WO2012080730. The applicant of the present disclosure has also disclosed in the patent WO2018095426 a selective TLR7 agonist with a pyrazoloheteroaryl derivative structure, which has good biological activity.

Although TLR7 agonists are used in cancer immunotherapy, many clinical trials are in progress, but most of them are not going well. The main reason is that the systemic administration of TLR7 agonist will bring strong side effects, such as lymphopenia, fever, headache and other cold-like symptoms. Imiquimod, as the only currently marketed TLR7 agonist, is also applied topically to reduce the toxic and side effects caused by systemic administration.

Based on the above considerations, based on the patent WO2018095426, the present disclosure further develops a prodrug of a novel structure TLR7 agonist, which can slowly release the active TLR7 agonist locally in the tumor. By changing the structure of the compound tail chain in the present disclosure, the speed at which the prodrug releases the active molecule can be adjusted to achieve the purpose of slowly releasing the active molecule, thereby prolonging the residence time of the drug in the tumor and reducing the systemic exposure of the drug. On the one hand, the number of injections can be reduced, and on the other hand, the slow release in the tumor tissue can reduce the toxic and side effects caused by system exposure.

Summary of the invention

The purpose of the present disclosure is to provide a compound represented by the general formula (I):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

Y is selected from -C (O) O-, -C (O) NH-, -C (O)-, -C (O) (CH ₂ ) _s- , -C (O) NH (CH ₂ ) _s- Or -C (O) (CR ^a R ^b ) _s NHC (O) CH _2- ;

R ^a and R ^{b are the} same or different, and are each independently selected from a hydrogen atom, halogen, alkyl, haloalkyl, hydroxy, or hydroxyalkyl;

R ^{0 is} selected from-(CH ₂ CH ₂ O) _n- , alkylene,-(CH ₂ CH ₂ O) _n -alkylene-,-(OCH ₂ CH ₂ ) _n -O- or-(OCH ₂ CH ₂ ) _n -O-alkylene-;

R is selected from _{-CH 3, -CH 2 CH 2 -} , - CH 2 -, - CH- , or C;

Ring A is selected from cycloalkyl, heterocyclyl, aryl and heteroaryl;

G is CH or N;

X ¹ is alkylene or S (O) _m , wherein the alkylene is optionally selected from halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro , Cycloalkyl and heterocyclic groups are substituted by one or more substituents;

L ^{1 is} selected from -NR ^4- , -O-, -S-, -C (O)-, -C (O) -OR ⁴ , -S (O) _m- , -N (R ⁴ ) C (O )-, -C (O) N (R ⁴ )-, -N (R ⁴ ) S (O) _2- , -S (O) ₂ N (R ⁴ )-and covalent bonds;

R ^{1 is} selected from alkyl, alkoxy, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkane The group, heterocyclic group, aryl group and heteroaryl group are each independently selected from alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, One of heterocyclic group, aryl group, heteroaryl group, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ Or substituted by multiple substituents;

R ^{2 is the} same or different and is independently selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aromatic Group and heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group are each independently selected from the group consisting of alkyl group, alkoxy group, halogen, halogenated alkyl group, hydroxyl group, hydroxyl group Alkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ One or more substituents in R ⁷ and -C (O) NR ⁶ R ⁷ are substituted;

L ² is an alkylene group or a covalent bond, wherein the alkylene group is optionally selected from alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cyclic Alkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ Substituted by one or more substituents in

R ^{3 is} selected from haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ , wherein the cycloalkyl, heterocyclic, aryl and heteroaryl groups are each independently selected from Group, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, heteroaryl, -OR ⁸ , -C (O) R ^8. -S (O) _m R ⁸ , -NR ⁹ R ¹⁰ and -C (O) NR ⁹ R ¹⁰ are substituted by one or more substituents;

R ^{4 is} selected from hydrogen atom, alkyl group, haloalkyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

R ^{5 is} selected from hydrogen atom, alkyl group, haloalkyl group, amino group, hydroxyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

R ⁶ and R ^{7 are the} same or different, and are each independently selected from a hydrogen atom, alkyl group, haloalkyl group, cycloalkyl group, heterocyclic group, aryl group, heteroaryl group, -C (O) R ⁸ , -S ( O) _m R ⁸ and -C (O) NR ⁹ R ¹⁰ ; wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups are each independently selected from alkyl, alkoxy Substituted by one or more substituents in the group, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl;

Alternatively, the R ⁶ and R ⁷ together with the connected nitrogen atom form a heterocyclic group, wherein the heterocyclic group may contain 1 to 2 same or different options in addition to 1 nitrogen atom. Heteroatoms from N, O and S, and the heterocyclic group is optionally selected from alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, hetero Substituted by one or more substituents in the cyclic group, aryl group and heteroaryl group;

R ^{8 is} selected from hydrogen atom, alkyl group, haloalkyl group, amino group, hydroxyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group;

R ⁹ and R ^{10 are the} same or different, and are each independently selected from a hydrogen atom, an alkyl group, a halogenated alkyl group, an amino group, a hydroxyl group, a cycloalkyl group, a heterocyclic group, an aryl group, and a heteroaryl group;

t is 0, 1, 2, 3 or 4;

s is an integer from 1 to 6;

n is an integer from 0 to 460; n is an average;

q is 1, 2, 3, or 4; and

m is 0, 1, or 2.

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein the ring A is selected from phenyl and pyridyl, wherein the pyridyl is preferably selected from

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein X ¹ is an alkylene group; L ^{1 is} selected from -O-, -NR ^4- , -C ( O)-and -C (O) N (R ⁴ )-, R ⁴ is a hydrogen atom or an alkyl group.

In other embodiments of the present disclosure, the compound represented by general formula (I) is a compound represented by general formula (II) or general formula (III):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

Y, R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in the general formula (I).

In other embodiments of the present disclosure, the compound represented by the general formula (I) is a compound represented by the general formula (IV):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

Y is selected from -C (O) O- or ^{-C (O) (CR a R} b) s NHC (O) CH 2 -; R a and R ^{b are} identical or different and are each independently selected from hydrogen atoms or alkoxy Base, s is an integer from 1 to 3;

R ^{0 is} selected from-(CH ₂ CH ₂ O) _n- , alkylene,-(CH ₂ CH ₂ O) _n -alkylene-,-(OCH ₂ CH ₂ ) _n -O- or-(OCH ₂ CH ₂ ) _n -O-alkylene-;

q, R is as defined in the general formula (I);

n is an average of about 1 to about 460.

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein the R ³ is a heterocyclic group, and the heterocyclic group is optionally selected from alkyl, alkoxy One or more substituents in the group, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclic, aryl and heteroaryl are substituted.

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein R ³ is -NR ⁶ R ⁷ , R ⁶ and R ⁷ together with the nitrogen atom to form a heterocyclic group, Wherein the heterocyclic group contains not only 1 nitrogen atom, but also optionally contains 1 or 2 heteroatoms selected from N, O and S which are the same or different, and the heterocyclic group is optionally selected from The alkyl group, alkoxy group, halogen, amino group, cyano group, nitro group, hydroxyl group, hydroxyalkyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group are substituted by one or more substituents.

In other embodiments of the present disclosure, the compound represented by general formula (I) is a compound represented by general formula (IIA) or general formula (IIIA):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

R ⁶ and R ⁷ together with the connected nitrogen atom form a heterocyclic group, wherein the heterocyclic group contains 1 nitrogen atom, and optionally contains 1 or 2 same or different selected from N, O And S heteroatoms, and the heterocyclic group is optionally selected from alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aromatic Substituted by one or more substituents in the group and heteroaryl.

R ⁰ , G, L ² , R ¹ , R ² and t are as defined in the general formula (I).

In other embodiments of the present disclosure, the compound represented by the general formula (IIA) is a compound represented by the general formula (IIAa):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

n is an integer from 0 to 20;

p is an integer from 0 to 18; and n and p are not 0 at the same time;

L ² , R ¹ , R ² , R ⁶ , R ⁷ and t are as defined in the general formula (IIA).

In other embodiments of the present disclosure, the compound represented by the general formula (IIIA) is a compound represented by the general formula (IIIAa):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

n is an integer from 1 to 10;

L ² , R ¹ , R ² , R ⁶ , R ⁷ and t are as defined in the general formula (IIIA).

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein the G is N.

In other embodiments of the present disclosure, the compound represented by the general formula (I) is a compound represented by the general formula (V):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

Y, R ¹ , R ⁰ , q, R are as defined in the general formula (IV).

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein R ¹ is an alkyl group, wherein the alkyl group is optionally substituted with one or more alkoxy groups.

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein L ² is an alkylene group; R ^{2 is the} same or different, and each is independently a hydrogen atom or a halogen.

In other embodiments of the present disclosure, the compound represented by general formula (I) is a compound represented by general formula (IIB) or general formula (IIIB):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

Y and R ⁰ are as defined in the general formula (I).

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein the Y is -C (O) O- or -C (O)-.

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein the R ⁰ is-(CH ₂ CH ₂ O) _n- (CH ₂ ) _p- , n is 0 Integer to 20, p is an integer from 0 to 18, n and p are not 0 at the same time; preferably-(CH ₂ CH ₂ O) _n -,-(CH ₂ ) _p -or -CH ₂ CH ₂ O- ( CH ₂ ) _p- , n is 1, or an integer of 8 to 14, and p is an integer of 8 to 18.

In other embodiments of the present disclosure, the compound represented by the general formula (IIB) is a compound represented by the general formula (IIC):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

n is an integer from 0 to 20;

p is an integer from 0 to 18; and n and p are not 0 at the same time.

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein Y is -C (O) O-, R ^{0 is} selected from-(CH ₂ CH ₂ O) _n -, Alkylene or-(CH ₂ CH ₂ O) _n -alkylene-, n is as defined in the general formula (I).

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein said Y is -C (O) (CR ^a R ^b ) _s NHC (O) CH _2- , R ⁰ is _{- (OCH 2 CH 2) n} -O- or _{- (OCH 2 CH 2) n} -O- alkylene ^{-, R a, R b,} n, s in the general formula (I) as defined above.

In other embodiments of the present disclosure, the compound represented by the general formula (I) is a compound represented by the general formula (VI):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

Among them, R ¹ , q, R are as defined in the general formula (IV);

Exemplary embodiment 1 of general formula (VI): when R is CH ₃ , q is 1; exemplary embodiment 2: when R is C, q is 4.

In other embodiments of the present disclosure, the compound represented by the general formula (I) is a compound represented by the general formula (VII):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

Wherein, R ¹ , q, R are as defined in the general formula (IV); p is an integer of 0 to 18; and n and p are not 0 at the same time;

Exemplary embodiment 1 of general formula (VII): when R is CH ₃ , q is 1; exemplary embodiment 2: when R is CH, q is 2.

In other embodiments of the present disclosure, the compound represented by the general formula (I), wherein n is an average of about 8 to about 500, preferably an average of about 10 to about 460, more It is preferably an average of about 10 to about 300.

In other embodiments of the present disclosure, the compound represented by the general formula (IIB) is a compound represented by the general formula (IIIC):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof,

among them:

n is an integer from 1 to 10.

Typical compounds of this disclosure include, but are not limited to:

a: The synthesis process and activity data of active molecule 1 are shown in WO2018095426.

Or tautomers, meso, racemates, enantiomers, diastereomers, or mixtures thereof or pharmaceutically acceptable salts thereof;

n is an average of 8 to 460, preferably an average of 10-460, and more preferably an average of 10-300.

Another aspect of the present disclosure relates to a method of preparing a compound represented by general formula (II), the method comprising:

The compound of formula (IA) and the compound of formula (IB) are reacted to obtain the compound of formula (II);

among them:

W is a leaving group, preferably halogen or

Y is -C (O) O- or -C (O)-;

R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in the general formula (II).

Another aspect of the present disclosure relates to a method of preparing a compound represented by general formula (III), the method including:

2 molecules of the compound of the general formula (IA) react with 1 molecule of the compound of the general formula (IC) to obtain the compound of the general formula (III);

among them:

W is a leaving group, preferably halogen or

Y is -C (O) O- or -C (O)-;

R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in the general formula (III).

Another aspect of the present disclosure relates to a method of preparing a compound represented by general formula (IIA), the method including:

The compound of general formula (II-1) and the compound of general formula (II-2) are reacted to obtain the compound of general formula (IIA);

among them:

W is a leaving group, preferably halogen or

R ⁰ , G, L ² , R ¹ , R ² , R ⁶ , R ⁷ and t are as defined in the general formula (IIA).

Another aspect of the present disclosure relates to a method of preparing a compound represented by general formula (IIIA), the method including:

2 molecules of the compound of the general formula (II-1) and 1 molecule of the compound of the general formula (III-1) are reacted to obtain the compound of the general formula (IIIA);

among them:

W is a leaving group, preferably halogen or

R ⁰ , G, L ² , R ¹ , R ² , R ⁶ , R ⁷ and t are as defined in the general formula (IIIA).

Another aspect of the present disclosure relates to a method of preparing a compound represented by general formula (IIB), the method including:

The compound of general formula (IIB-1) and the compound of general formula (IB) are reacted to obtain the compound of general formula (IIB);

among them:

W is a leaving group, preferably halogen or

Y and R ⁰ are as defined in the general formula (IIB).

Another aspect of the present disclosure relates to a method of preparing a compound represented by general formula (IIIB), the method including:

2 molecules of the compound of the general formula (IIB-1) and 1 molecule of the compound of the general formula (IC) are reacted to obtain the compound of the general formula (IIIB);

among them:

W is a leaving group, preferably halogen or

Y and R ⁰ are as defined in the general formula (IIIB).

Another aspect of the present disclosure relates to a method of preparing a compound represented by general formula (VI), the method comprising:

The compound of general formula (VI-A) and the compound of (VI-B) react under basic conditions to obtain the compound of general formula (VI);

Among them, R ¹ , R and q are as defined in the general formula (VI); p is preferably 0 or 1.

The reagents for providing alkaline conditions in the above reaction include organic bases and inorganic bases, and the organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyllithium, and diisopropyl Lithium amide, lithium bistrimethylsilylamine, potassium acetate, sodium tert-butoxide, potassium tert-butoxide and sodium n-butoxide, the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, carbonic acid Potassium, potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1 , 4-dioxane, water or N, N-dimethylformamide.

Another aspect of the present disclosure relates to a pharmaceutical composition containing a therapeutically effective amount of a compound according to general formula (I) or a tautomer thereof, a racemate, a racemate , Enantiomers, diastereomers, or mixtures thereof or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, is used in the preparation of a medicament for agonizing TLR7.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or Use of a pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, in the preparation of a medicament for treating or preventing TLR7 mediated diseases.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or The use of a pharmaceutically acceptable salt or a pharmaceutical composition containing the same in the preparation of a medicament for treating or preventing tumors; wherein the tumor is preferably selected from melanoma, lung cancer, small cell lung cancer, non-small cell lung cancer, liver cells Cancer, basal cell carcinoma, renal cell carcinoma, myeloma, breast cancer, ovarian cancer, colon cancer, rectal cancer, prostate cancer, brain tumor, esophageal cancer, colorectal cancer, stomach cancer, liver cancer, pancreatic cancer, colorectal cancer, kidney Cancer, skin cancer, neuroblastoma, sarcoma, osteochondroma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck cancer, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, Huo Chikin's disease, adrenal cortical tumors, thyroid tumors, ureteral tumors, bladder cancer, gallbladder cancer, cholangiocarcinoma, chorionic epithelial cancer, and pediatric tumors.

The present disclosure further relates to a compound represented by general formula (I) or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof, or Use of a pharmaceutically acceptable salt or a pharmaceutical composition containing the same in the preparation of a medicament for treating infections caused by viruses selected from the group consisting of: dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis Viruses, tick-borne encephalitis virus, Kunjin virus, Murray valley encephalitis virus, Saint Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus, HIV, HBV, HCV, HPV, RSV, SARS and influenza viruses.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing it, is used as a medicine.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing it, which is used for agonizing TLR7.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, for use in the treatment or prevention of TLR7 mediated diseases.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, for the treatment or prevention of tumors; wherein the tumors are preferably selected from melanoma, lung cancer, small cell lung cancer, non-small cell lung cancer, hepatocellular carcinoma, basal cell carcinoma , Renal cell carcinoma, myeloma, breast cancer, ovarian cancer, colon cancer, rectal cancer, prostate cancer, brain tumor, esophageal cancer, colorectal cancer, gastric cancer, liver cancer, pancreatic cancer, colorectal cancer, kidney cancer, skin cancer, Neuroblastoma, sarcoma, osteochondroma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck cancer, multiple myeloma, malignant lymphoma, polycythemia vera, leukemia, Hodgkin's disease, adrenal gland Cortical tumors, thyroid tumors, ureteral tumors, bladder cancer, gallbladder cancer, cholangiocarcinoma, chorionic epithelial cancer, and pediatric tumors.

The present disclosure further relates to compounds represented by general formula (I) or tautomers, meso, racemates, enantiomers, diastereomers or mixtures thereof, or A pharmaceutically acceptable salt, or a pharmaceutical composition containing the same, for the treatment of infections caused by viruses selected from dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis virus, tick-borne encephalitis Viruses, Kunjin virus, Murray Valley encephalitis virus, Saint Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus, HIV, HBV, HCV, HPV, RSV, SARS and influenza virus .

The present disclosure further relates to a method of stimulating TLR7, which method comprises administering to a patient in need thereof a therapeutically effective dose of a compound represented by the general formula (I) of the present disclosure or a tautomer, a racemate, an exosome Racemate, enantiomer, diastereomer or a mixture thereof, or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the same.

The present disclosure further relates to a method of treating or preventing TLR7-mediated diseases, which includes administering to a desired patient a therapeutically effective amount of a compound represented by the general formula (I) or a tautomer, a racemate, an exosome Racemate, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same.

The present disclosure further relates to a method of treating or preventing tumors, which comprises administering to a desired patient a therapeutically effective amount of a compound represented by general formula (I) or a tautomer thereof, a racemate, a racemate, Enantiomers, diastereomers, or mixtures thereof, or pharmaceutically acceptable salts or pharmaceutical compositions containing them; wherein the tumor is preferably selected from melanoma, lung cancer, small cell lung cancer, non- Small cell lung cancer, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, breast cancer, ovarian cancer, colon cancer, rectal cancer, prostate cancer, brain tumor, esophageal cancer, colorectal cancer, gastric cancer, liver cancer, pancreatic cancer , Colorectal cancer, kidney cancer, skin cancer, neuroblastoma, sarcoma, osteochondroma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck cancer, multiple myeloma, malignant lymphoma, true red blood cells Hyperplasia, leukemia, Hodgkin's disease, adrenal cortical tumors, thyroid tumors, ureteral tumors, bladder cancer, gallbladder cancer, cholangiocarcinoma, chorionic epithelial cancer, and pediatric tumors.

The present disclosure further relates to a method of treating infections caused by viruses, which includes administering to a desired patient a therapeutically effective amount of a compound represented by general formula (I) or a tautomer, a racemate, a racemate Isomer, enantiomer, diastereomer, or a mixture thereof, or a pharmaceutically acceptable salt or pharmaceutical composition containing the same, wherein the virus is selected from the group consisting of: dengue virus, yellow fever virus, West Nile virus, Japanese encephalitis virus, tick-borne encephalitis virus, Kunjin virus, Murray valley encephalitis virus, Saint Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus, HIV , HBV, HCV, HPV, RSV, SARS and influenza viruses.

The active compound can be prepared in a form suitable for administration by any suitable route. The active compound is preferably in unit dosage form, or in such a way that the patient can self-administer it in a single dose. The unit dosage of the disclosed compound or composition may be expressed in the form of tablets, capsules, cachets, bottled syrups, powders, granules, lozenges, suppositories, regenerated powders or liquid preparations.

The dosage of the compound or composition used in the treatment methods of the present disclosure will generally vary with the severity of the disease, the weight of the patient, and the relative efficacy of the compound. However, as a general guide, a suitable unit dose may be 0.1-1000 mg.

In addition to the active compound, the pharmaceutical composition of the present disclosure may contain one or more excipients selected from the following ingredients: fillers (diluents), binders, wetting agents, disintegrating agents, or excipients Wait. Depending on the method of administration, the composition may contain from 0.1 to 99% by weight of active compound.

The pharmaceutical composition containing the active ingredient may be in a form suitable for oral administration, such as tablets, dragees, lozenges, water or oil suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or Elixir. Oral compositions can be prepared according to any method known in the art for preparing pharmaceutical compositions. Such compositions may contain binders, fillers, lubricants, disintegrating agents or pharmaceutically acceptable wetting agents, etc. The composition may also contain one or more ingredients selected from the group consisting of sweeteners, flavoring agents, coloring agents and preservatives to provide pleasing and palatable pharmaceutical preparations.

Aqueous suspensions contain the active substance and excipients suitable for the preparation of aqueous suspensions for mixing. The aqueous suspension may also contain one or more preservatives, one or more coloring agents, one or more flavoring agents, and one or more sweeteners.

Oil suspensions can be prepared by suspending the active ingredient in vegetable oil. The oil suspension may contain a thickener. The above sweeteners and flavoring agents can be added to provide a palatable preparation.

The pharmaceutical composition may also be dispersible powders and granules used to prepare an aqueous suspension to provide the active ingredient by adding one or more of a dispersant, wetting agent, suspending agent or preservative by adding water. Other excipients such as sweeteners, flavoring agents and coloring agents can also be added. These compositions are preserved by adding antioxidants such as ascorbic acid.

The pharmaceutical composition of the present disclosure may also be in the form of an oil-in-water emulsion.

The pharmaceutical composition may be in the form of a sterile injectable aqueous solution. Acceptable solvents or solvents that can be used are water, Ringer's solution, and isotonic sodium chloride solution. The sterile injectable preparation may be a sterile injectable oil-in-water microemulsion in which the active ingredient is dissolved in the oily phase. For example, the active ingredient is dissolved in a mixture of soybean oil and lecithin. The oil solution is then added to a mixture of water and glycerin to form a microemulsion. The injection solution or microemulsion can be injected into the patient's bloodstream by local injection. Alternatively, it is best to administer the solution and microemulsion in a manner that maintains a constant circulating concentration of the disclosed compound. To maintain this constant concentration, a continuous intravenous drug delivery device can be used. An example of such a device is the Deltec CADD-PLUS.TM. 5400 type intravenous pump.

The pharmaceutical composition may be in the form of a sterile injectable water or oil suspension for intramuscular and subcutaneous administration. The suspension can be formulated according to known techniques using those suitable dispersing or wetting agents and suspending agents described above. The sterile injectable preparation may also be a sterile injectable solution or suspension prepared in a parenterally acceptable non-toxic diluent or solvent. In addition, sterile fixed oil can be conveniently used as a solvent or suspending medium.

The compounds of the present disclosure can be administered in the form of suppositories for rectal administration. These pharmaceutical compositions can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid in the rectum and therefore will dissolve in the rectum to release the drug. Such substances include cocoa butter, glycerinated gelatin, hydrogenated vegetable oils, polyethylene glycols of various molecular weights and mixtures of fatty acid esters of polyethylene glycols.

As is well known to those skilled in the art, the dosage of a drug depends on many factors, including but not limited to the following factors: the activity of the specific compound used, the age of the patient, the weight of the patient, the health of the patient, the behavior of the patient , The patient's diet, administration time, administration method, excretion rate, drug combination, etc. In addition, the optimal treatment method such as the mode of treatment, the daily dosage of the general formula (I) or pharmaceutically acceptable salts The type can be verified according to the traditional treatment plan.

Detailed description of the invention

Unless stated to the contrary, the terms used in the specification and claims have the following meanings.

The term "alkyl" refers to a saturated aliphatic hydrocarbon group, which is a linear or branched group containing 1 to 20 carbon atoms, preferably an alkyl group containing 1 to 12 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1 , 2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2- Methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3 -Dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2 -Methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethyl Amyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2 , 5-dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4-ethyl Hexyl, 2-methyl-2-ethylpentyl, 2-methyl-3-ethylpentyl, n-nonyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethyl Ghexyl, 2,2- Diethylpentyl, n-decyl, 3,3-diethylhexyl, 2,2-diethylhexyl, and its various branched isomers, etc. More preferred are lower alkyl groups containing 1 to 6 carbon atoms, non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl Group, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethyl Butyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl Group, 2,3-dimethylbutyl, etc. The alkyl group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any usable connection point. The substituent is preferably independently selected from alkyl, alkenyl, alkynyl, Alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy , Cycloalkylthio, heterocycloalkylthio, oxo, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ One or more substituents in R ⁷ are substituted.

The term "alkylene" refers to a saturated straight-chain or branched-chain aliphatic hydrocarbon group having 2 residues derived from the removal of two hydrogen atoms from the same carbon atom of the parent alkane or two different carbon atoms, which is Straight or branched chain groups containing 1 to 30 carbon atoms, preferably containing 1 to 18 carbon atoms, more preferably 1 to 12 carbon atom alkylene groups, most preferably 1 to 6 carbon atom subalkylene groups alkyl. Non-limiting examples of alkylene include, but are not limited to, methylene (-CH _2- ), 1,1-ethylene (-CH (CH ₃ )-), 1,2-ethylene (-CH ₂ CH ₂ )-, 1,1-propylene (-CH (CH ₂ CH ₃ )-), 1,2-propylene (-CH ₂ CH (CH ₃ )-), 1,3-propylene (-CH ₂ CH ₂ CH _2- ), 1,4-butylene (-CH ₂ CH ₂ CH ₂ CH ₂ -),-(CH ₂ ) ₉ -,-(CH ₂ ) _17- , etc. The alkylene group may be substituted or unsubstituted. When substituted, the substituent may be substituted at any available connection point, and the substituent is preferably independently selected from alkyl, alkenyl, alkynyl. , Alkoxy, alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy Group, cycloalkylthio group, heterocycloalkylthio group, oxo group, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ is substituted by one or more substituents.

The term "alkenyl" refers to a hydrocarbon group formed by one or fewer hydrogen atoms in an olefin molecule. The alkenyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from a hydrogen atom, an alkyl group, an alkoxy group, a halogen, a halogenated alkyl group, a hydroxyl group, Hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ are substituted by one or more substituents;

The term "alkynyl" refers to a hydrocarbon compound containing a carbon-carbon triple bond in the molecule. The alkynyl group may be substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from a hydrogen atom, an alkyl group, an alkoxy group, a halogen, a halogenated alkyl group, a hydroxyl group, Hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ are substituted by one or more substituents.

The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent. The cycloalkyl ring contains 3 to 20 carbon atoms, preferably 3 to 12 carbon atoms, preferably 3 to 10 carbon atoms The carbon atoms more preferably contain 3 to 6 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, cycloheptyl, cycloheptatriene Radicals, cyclooctyl, etc .; polycyclic cycloalkyls include spiro, fused and bridged cycloalkyls.

The term "amino protecting group" is to keep the amino group unchanged during the reaction of other parts of the molecule, and protect the amino group with a group that can be easily removed. Non-limiting examples include tert-butoxycarbonyl, acetyl, benzyl, allyl, p-methoxybenzyl and the like. These groups may be optionally substituted with 1-3 substituents selected from halogen, alkoxy or nitro. The amino protecting group is preferably p-methoxybenzyl.

The term "heterocyclic group" refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic hydrocarbon substituent, which contains 3 to 20 ring atoms, wherein one or more ring atoms are selected from nitrogen, oxygen, or S (O) _m (where m is an integer from 0 to 2), but does not include the ring portion of -OO-, -OS-, or -SS-, and the remaining ring atoms are carbon. It preferably contains 3 to 12 ring atoms, of which 1 to 4 are heteroatoms; more preferably 3 to 10 ring atoms, of which 1 to 4 are heteroatoms; more preferably 5 to 6 ring atoms; of which 1 to 3 These are heteroatoms. Non-limiting examples of monocyclic heterocyclic groups include pyrrolidinyl, tetrahydropyranyl, 1,2.3.6-tetrahydropyridinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, High piperazinyl and so on. Polycyclic heterocyclic groups include spiro, fused and bridged heterocyclic groups.

The heterocyclic ring includes a ring fused to an aryl, heteroaryl or cycloalkyl ring, wherein the ring connected to the parent structure is a heterocyclic group, and non-limiting examples include:

Heterocyclic groups include optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, Alkylthio, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkane Sulfur, heterocycloalkylthio, oxo, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ Substituted by one or more substituents.

The term "aryl" refers to a 6 to 14-membered all-carbon monocyclic or fused polycyclic (ie, rings that share adjacent pairs of carbon atoms) groups with a conjugated π-electron system, preferably 6 to 10 members, such as benzene And naphthyl. The aryl ring may be fused to a heteroaryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is an aryl ring, and non-limiting examples thereof include:

Aryl includes substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio , Alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio, One or more substituents in heterocycloalkylthio, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ Replaced by.

The term "heteroaryl" refers to a heteroaromatic system containing 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms are selected from oxygen, sulfur, and nitrogen. The heteroaryl group is preferably 5- to 10-membered, more preferably 5- or 6-membered, such as furyl, thienyl, pyridyl, pyrrolyl, N-alkylpyrrolyl, pyrimidinyl, pyrazinyl, pyridazinyl, Imidazolyl, pyrazolyl, tetrazolyl, etc. The heteroaryl ring may be fused to an aryl, heterocyclic or cycloalkyl ring, wherein the ring connected to the parent structure is a heteroaryl ring, and non-limiting examples thereof include:

Heteroaryl includes optionally substituted or unsubstituted, when substituted, the substituent is preferably one or more of the following groups, which are independently selected from alkyl, alkenyl, alkynyl, alkoxy, alkylthio Group, alkylamino, halogen, thiol, hydroxyl, nitro, cyano, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heterocycloalkoxy, cycloalkylthio , One or more of heterocycloalkylthio, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ Substituted.

The term "alkoxy" refers to -O- (alkyl) and -O- (unsubstituted cycloalkyl), where alkyl is as defined above. Non-limiting examples of alkoxy groups include: methoxy, ethoxy, propoxy, butoxy, cyclopropoxy, cyclobutoxy, cyclopentyloxy, cyclohexyloxy. The alkoxy group may be optionally substituted or unsubstituted. When substituted, the substituent is preferably one or more of the following groups, which are independently selected from halogen, alkyl, alkoxy, haloalkyl, hydroxy, One or more substituents in the hydroxyalkyl group, cyano group, amino group, nitro group, cycloalkyl group, heterocyclic group, aryl group, and heteroaryl group are substituted.

The term "leaving group" refers to an atom or functional group that is detached from a larger molecule in a chemical reaction.

The term "haloalkyl" refers to an alkyl group substituted with one or more halogens, where alkyl is as defined above.

The term "hydroxyl" refers to the -OH group.

The term "hydroxyalkyl" refers to an alkyl group substituted with a hydroxy group, where alkyl is as defined above.

The term "halogen" refers to fluorine, chlorine, bromine or iodine.

The term "amino" means -NH _2.

The term "cyano" refers to -CN.

The term "nitro" refers to -NO ₂ .

The term "oxo" refers to = O.

-(CH ₂ CH ₂ O) n- or-(OCH ₂ CH ₂ ) nO- in the general formula of the present disclosure is a water-soluble non-peptide polymer composed of repeating monomers.

"Water-soluble non-peptide polymer" refers to a polymer that is at least 35% (by weight) soluble in water at room temperature, preferably greater than 70% (by weight), and more preferably greater than 95% (by weight) soluble. Typically, an unfiltered aqueous formulation of a "water soluble" polymer transmits at least 75% (more preferably at least 95%) of the amount of light transmitted by the same solution after filtration. However, it is most preferred that the water-soluble polymer is at least 95% (by weight) water-soluble or completely water-soluble. With regard to being "non-peptide", the polymer is non-peptidic when it has less than 35% (by weight) of amino acid residues.

The terms "monomer", "monomer subunit" and "monomer unit" are used interchangeably herein and refer to one of the basic structural units of the polymer. In the case of homopolymers, a single repeating structural unit forms the polymer. In the case of copolymers, two or more structural units appear repeatedly—in a pattern or randomly—to form the polymer. The preferred polymer is a homopolymer. The water-soluble, non-peptide polymer includes one or more monomers attached in series to form a monomer chain. The polymer may be formed from a single monomer type (ie, homopolymer type) or two or three monomer types (ie, is a copolymer type).

As used herein, a "polymer" is a molecule having from about 2 to about 2000 or more (eg, from about 2 to about 4000) monomers. Specific polymers include those with various geometries (eg, linear, branched, or fork-shaped) as described in more detail below.

"PEG" and "polyethylene glycol" as used herein are meant to include any water-soluble poly (ethylene oxide). Unless otherwise specified, "PEG polymer" or any polyethylene glycol is a substance in which substantially all (preferably all) monomer subunits are ethylene oxide subunits, however, the polymer may contain Different end-capping moieties or functional groups are used, for example, for conjugation. PEG polymer can include one of the following two structures: "-(CH ₂ CH ₂ O) n-" or "-(CH ₂ CH ₂ O) n- ₁ CH ₂ CH _2- ", depending on the terminal Whether one or more of the oxygen is replaced (eg during synthetic conversion). As mentioned above, for these PEG polymers, the variable (n) (ie, the number of repeating units) ranges from about 2 to 2000, or from 2 to 4000, and the end groups and framework of the entire PEG can vary. When PEG further includes a functional group for attachment to, for example, a small molecule drug, the functional group does not cause the formation of an oxygen-oxygen bond (-OO-, a peroxide linkage) when covalently attached to the PEG polymer .

The terms "end-capped" or "end-capped" are used interchangeably herein to refer to the ends or ends of a polymer having a capped portion. Typically, although not required, the end-capping moiety includes a hydroxyl group or a C _1-20 alkoxy or basic aryloxy (alkaaryloxy) group. Therefore, examples of the end-capping portion include alkoxy groups (eg, methoxy and ethoxy), benzyloxy, and aryl, heteroaryl, cyclic, heterocyclic, and the like. In addition, the saturated, unsaturated, substituted, and unsubstituted forms of each of the foregoing groups are envisioned. In addition, the end-capping group can also be a silane. The end-capping group can also advantageously contain a detectable label. When the polymer has a capping group containing a detectable label, the amount or position of the polymer and / or the moiety of interest (eg, an activator) to which the polymer is coupled can be determined by using a suitable detector. Such labels include, but are not limited to: fluorescent agents, chemiluminescent agents, moieties used in enzyme labeling, colorimetric moieties (eg, dyes), metal ions, radioactive moieties, and the like. Suitable detectors include photometers, films, spectrometers, etc. In addition, the end-capping group may include a targeting moiety.

The term "targeting moiety" refers to a molecular structure that helps localize the conjugate to the targeted region (eg, helps enter cells or bind receptors). Preferably, the targeting moiety includes vitamins, antibodies, antigens, receptors, DNA, RNA, sialylated Lewis X antigen, hyaluronic acid, carbohydrates, cell-specific lectins, steroids or steroid derivatives, RGD peptides, cells Surface receptor ligands, serum components, or combined molecules directed against different intracellular or extracellular receptors. The targeting moiety can also include lipids or phospholipids. Exemplary phospholipids include, but are not limited to, phosphatidylcholines, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, and phosphatidylethanolamine. These lipids can be in the form of micelles or liposomes and the like. The targeting moiety can further include a detectable label, or alternatively the detectable label can function as a targeting moiety. When the polymer conjugate has a targeting group that includes a detectable label, a suitable detector can be used to determine the amount of the polymer and / or the moiety to which the polymer is conjugated (eg, an activator) and / or Or distribution / location. Such labels include, but are not limited to: fluorescent agents, chemiluminescent agents, moieties used in enzyme labeling, colorimetric substances (such as dyes), metal ions, radioactive moieties, gold particles, quantum dots, and the like.

In the context of water-soluble polymers such as PEG, molecular weight can be expressed as number average molecular weight or weight average molecular weight. Unless otherwise indicated, all references to molecular weight herein refer to weight average molecular weight. Both molecular weight determinations (number average and weight average molecular weight) can be measured using gel permeation chromatography or other liquid chromatography techniques. Other methods for measuring molecular weight values can also be used, such as the use of end-group analysis or the measurement of colligative properties (eg, freezing point depression, boiling point elevation, or osmotic pressure) to determine the number average molecular weight or the use of light scattering techniques, Ultracentrifugation or viscometry is used to determine the weight average molecular weight. PEG polymers are typically polydisperse (ie, the number average molecular weight of these polymers is not equal to the weight average molecular weight), having preferably less than about 1.2, more preferably less than about 1.15, still more preferably less than about 1.10, yet still more preferably Low polydispersity values less than about 1.05 and most preferably less than about 1.03.

With regard to the geometry or overall structure of a polymer, "branched" refers to a polymer having two or more polymer "arms" extending from a branch point.

With regard to the geometry or overall structure of the polymer, "forked" refers to a polymer having two or more functional groups (typically through one or more atoms) extending from the branch point.

"Branch point" refers to a divergence point that includes one or more atoms, at which point the polymer branches or branches from a linear structure into one or more additional arms.

These multi-arm polymer conjugates contain water-soluble non-peptide polymers. A large number of polymers can be used, and the structure provided herein is not limited with regard to the type of water-soluble polymer (eg, polyethylene oxide or polyoxazoline) or size (eg, from 2 to 4,000 monomer sizes) .

Regarding types, water-soluble non-peptide polymers are understood as a series of repeating monomers, where the type of one or more monomers determines the type of water-soluble non-peptide polymers. Exemplary monomers include, but are not limited to, alkylene oxides, such as ethylene oxide or propylene oxide; enols, such as vinyl alcohol, 1-propenol, or 2-propenol; vinyl pyrrolidone; hydroxyalkyl methacrylamide, and Hydroxyalkyl methacrylate, wherein, in each case, the alkyl is preferably methyl; alpha-hydroxy acid, such as lactic acid or glycolic acid; phosphazene, oxazoline, carbohydrate (such as monosaccharide), Sugar alcohols (such as mannitol); and N-acryloyl morpholine. In one or more embodiments, the water-soluble non-peptide polymer is a copolymer of two monomer types selected from this group, or more preferably a homopolymer of one monomer type selected from this group. With regard to copolymers including block copolymers, the two monomer types in the copolymer may be the same monomer type, for example, two alkylene oxides (such as ethylene oxide and propylene oxide).

Regarding size, the water-soluble non-peptide polymer may be relatively small, or the water-soluble non-peptide polymer may be relatively large.

With regard to water-soluble non-peptide polymers, in other words, with respect to each polymer arm in those embodiments where relatively small water-soluble non-peptide polymers are present, exemplary values of molecular weight include: less than about 2000; less than about 1500; Below about 1450; below about 1400; below about 1350; below about 1300; below about 1250; below about 1200; below about 1150; below about 1100; below about 1050; below about 1000; Below about 950; below about 900; below about 850; below about 800; below about 750; below about 700; below about 650; below about 600; below about 550; below about 500; Below about 450; below about 400; below about 350; below about 300; below about 250; below about 200; and below about 100 Daltons. Exemplary ranges of relatively small water-soluble non-peptide polymers include from about 100 to about 1400 Daltons; from about 100 to about 1200 Daltons; from about 100 to about 800 Daltons; from about 100 to about 500 Dalton; from about 100 to about 400 Daltons; from about 200 to about 500 Daltons; from about 200 to about 400 Daltons; from about 75 to 1000 Daltons; and from about 75 to about 750 Dalton.

For relatively small water-soluble non-peptide polymers, the number of monomers will generally fall within one or more of the following ranges: between 1 and about 30 (inclusive); between about 2 and about 25; Between about 2 and about 20; between about 2 and about 15; between about 2 and about 12; between about 2 and about 10. In some cases, the number of monomers in series in the polymer (and corresponding conjugate) is one of 1, 2, 3, 4, 5, 6, 7, or 8. In additional embodiments, the polymer (and corresponding conjugate) contains 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 monomers. In yet other embodiments, the polymer portion (and corresponding conjugate) in each polymer "arm" has 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 in series Monomers. Thus, for example, when the water-soluble non-peptide polymer arm contains-(OCH ₂ CH ₂ ) n-, "n" is an integer such that, in some embodiments, it is selected from 1, 2, 3, 4, 5 , 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29 and 30 , And can fall into one or more of the following ranges: between about 1 and about 25; between about 1 and about 20; between about 1 and about 15; between about 1 and about 12; between about 1 and about Between 10.

When the molecular weight of the total water-soluble non-peptide polymer in the conjugate is relatively large (eg, greater than 2,000 Daltons), the total molecular weight can fall within the range of 2,000 Daltons to about 150,000 Daltons. However, exemplary ranges include the following molecular weights: in the range from about 3,000 Daltons to about 120,000 Daltons; in the range from about 5,000 Daltons to about 110,000 Daltons; in the range from greater than 5,000 Daltons to about 100,000 Daltons, from about 6,000 Daltons to about 90,000 Daltons, from about 10,000 Daltons to about 85,000 Daltons, from more than 10,000 Daltons to about 85,000 Daltons Range from about 20,000 Daltons to about 85,000 Daltons, from about 53,000 Daltons to about 85,000 Daltons, from about 25,000 Daltons to about 120,000 Daltons Range, in the range from about 29,000 Daltons to about 120,000 Daltons, in the range from about 35,000 Daltons to about 120,000 Daltons, and in the range from about 40,000 Daltons to about 120,000 Daltons .

Exemplary molecular weights for relatively large water-soluble non-peptide polymers (for each polymer arm water-soluble non-peptide polymer in Formula I) include about 500 Daltons, about 750 Daltons, about 1,000 Daltons, About 1500 Daltons, about 2,000 Daltons, about 2,200 Daltons, about 2,500 Daltons, about 3,000 Daltons, about 4,000 Daltons, about 4,400 Daltons, about 4,500 Daltons, about 5,000 Dalton, about 5,500 Dalton, about 6,000 Dalton, about 7,000 Dalton, about 7,500 Dalton, about 8,000 Dalton, about 9,000 Dalton, about 10,000 Dalton, about 11,000 Dal Dayton, about 12,000 Daltons, about 13,000 Daltons, about 14,000 Daltons, about 15,000 Daltons, and about 20,000 Daltons.

Exemplary molecular weights for relatively large water-soluble non-peptide polymers (regarding the total polymer portion of the multi-arm conjugate) include, for example, about 20,000 Daltons, 22,500 Daltons, about 25,000 Daltons, about 30,000 Dalton, about 35,000 Dalton, about 40,000 Dalton, about 45,000 Dalton, about 50,000 Dalton, about 55,000 Dalton, about 60,000 Dalton, about 65,000 Dalton, about 70,000 Dal Dayton, and about 75,000 Daltons. Branched forms of water-soluble non-peptide polymers having any of the aforementioned total molecular weights can also be used in each polymer arm to provide multiple branched conjugates.

Therefore, regardless of whether a relatively small or relatively large water-soluble non-peptide polymer is used, when the water-soluble non-peptide polymer is poly (ethylene oxide), the polymer will contain multiple (OCH ₂ CH ₂ ) monomers [ Or (CH ₂ CH ₂ O) monomer, depending on how PEG is defined]. As used throughout the specification, the number of repeating units is identified by the subscript "n" in "(OCH ₂ CH ₂ ) n". Therefore, the value of (n) typically falls within one or more of the following ranges: from 1 to about 3400, from about 1 to about 2300, from about 1 to about 1000, from about 1 to about 500, from about 1 to about 350, from about 1 to about 300, from about 8 to about 300, from about 10 to about 300, from about 10 to about 280, from about 9 to about 280. For any given polymer of known molecular weight, it is possible to determine the number of repeating units (ie "n") by dividing the total weight average molecular weight of the polymer by the molecular weight of the repeating monomer.

Polyethylene oxide, a ring-opening polymer of ethylene oxide. It is a polyether polymer. The structure is the same as the glycol condensation polymer, also known as polyethylene glycol, the English abbreviation PEG.

"Optional" or "optionally" means that the subsequently described event or environment may but need not occur, and the description includes the occasion where the event or environment occurs or does not occur. For example, "heterocyclic group optionally substituted with an alkyl group" means that an alkyl group may but need not be present, and the description includes the case where the heterocyclic group is substituted with an alkyl group and the case where the heterocyclic group is not substituted with an alkyl group .

"Substituted" refers to one or more hydrogen atoms in a group, preferably up to 5, more preferably 1 to 3 hydrogen atoms are independently substituted with a corresponding number of substituents. It goes without saying that the substituents are only at their possible chemical positions, and those skilled in the art can determine (through experiment or theory) possible or impossible substitutions without undue effort. For example, an amino group or hydroxyl group having free hydrogen may be unstable when combined with a carbon atom having an unsaturated (eg, olefinic) bond.

"Pharmaceutical composition" means a mixture containing one or more compounds described herein or a physiological / pharmaceutically acceptable salt or prodrug thereof with other chemical components, and other components such as physiological / pharmaceutically acceptable carriers And excipients. The purpose of the pharmaceutical composition is to promote the administration to the living body, facilitate the absorption of the active ingredient and thus exert the biological activity.

"Pharmaceutically acceptable salt" refers to a salt of a compound of the present disclosure, such salt is safe and effective when used in mammals, and has due biological activity.

m and R ⁵ to R ⁷ are as defined in the compound of general formula (I).

Synthetic method of the disclosed compound

In order to accomplish the purpose of the present disclosure, the present disclosure adopts the following technical solutions:

Option One

The compound represented by the general formula (II) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or may be The preparation method of the medicinal salt includes the following steps:

The compound of general formula (IA) and the compound of general formula (IB) react under basic conditions to obtain the compound of general formula (II);

among them:

W is a leaving group, preferably halogen or

Y is -C (O) O- or -C (O)-;

R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in the general formula (II).

Reagents that provide basic conditions include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyllithium, and lithium diisopropylamide , Lithium bistrimethylsilylamine, potassium acetate, sodium tert-butoxide, potassium tert-butoxide and sodium n-butoxide, the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, Potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1 , 4-dioxane, water or N, N-dimethylformamide.

Option II

The compound represented by the general formula (III) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or may be The preparation method of the medicinal salt includes the following steps:

Two molecules of the compound of the general formula (IA) and one molecule of the compound of the general formula (IC) react under basic conditions to obtain the compound of the general formula (III);

among them:

W is a leaving group, preferably halogen or

Y is -C (O) O- or -C (O)-;

R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in the general formula (III).

Reagents that provide basic conditions include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyllithium, and lithium diisopropylamide , Lithium bistrimethylsilylamine, potassium acetate, sodium tert-butoxide, potassium tert-butoxide and sodium n-butoxide, the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, Potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1 , 4-dioxane, water or N, N-dimethylformamide.

third solution

The compound represented by the general formula (IIA) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or may be The preparation method of the medicinal salt includes the following steps:

The compound of general formula (II-1) and the compound of general formula (II-2) react under basic conditions to obtain the compound of general formula (IIA);

among them:

W is a leaving group, preferably halogen or

R ⁰ , G, L ² , R ¹ , R ² , R ⁶ , R ⁷ and t are as defined in the general formula (IIA).

Reagents that provide basic conditions include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyllithium, and lithium diisopropylamide , Lithium bistrimethylsilylamine, potassium acetate, sodium tert-butoxide, potassium tert-butoxide and sodium n-butoxide, the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, Potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1 , 4-dioxane, water or N, N-dimethylformamide.

Option Four

The compound represented by the general formula (IIIA) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or may be The preparation method of the medicinal salt includes the following steps:

Two molecules of the compound of the general formula (II-1) and one molecule of the compound of the general formula (III-1) react under basic conditions to obtain the compound of the general formula (IIIA);

among them:

W is a leaving group, preferably halogen or

R ⁰ , G, L ² , R ¹ , R ² , R ⁶ , R ⁷ and t are as defined in the general formula (IIIA).

Reagents that provide basic conditions include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyllithium, and lithium diisopropylamide , Lithium bistrimethylsilylamine, potassium acetate, sodium tert-butoxide, potassium tert-butoxide and sodium n-butoxide, the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, Potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1 , 4-dioxane, water or N, N-dimethylformamide.

Solution Five

The compound represented by the general formula (IIB) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or may be The preparation method of the medicinal salt includes the following steps:

The compound of general formula (IIB-1) and the compound of general formula (IB) react under basic conditions to obtain the compound of general formula (IIB);

among them:

W is a leaving group, preferably halogen or

Y and R ⁰ are as defined in the general formula (IIB).

Reagents that provide basic conditions include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyllithium, and lithium diisopropylamide , Lithium bistrimethylsilylamine, potassium acetate, sodium tert-butoxide, potassium tert-butoxide and sodium n-butoxide, the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, Potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1 , 4-dioxane, water or N, N-dimethylformamide.

Solution Six

The compound represented by the general formula (IIIB) of the present disclosure or its tautomer, meso, racemate, enantiomer, diastereomer, or a mixture thereof, or may be The preparation method of the medicinal salt includes the following steps:

Two molecules of the compound of the general formula (IIB-1) and one molecule of the compound of the general formula (IC) react under basic conditions to obtain the compound of the general formula (IIIB);

among them:

W is a leaving group, preferably halogen or

Y and R ⁰ are as defined in the general formula (IIIB).

Reagents that provide basic conditions include organic bases and inorganic bases. The organic bases include but are not limited to triethylamine, N, N-diisopropylethylamine, n-butyllithium, and lithium diisopropylamide , Lithium bistrimethylsilylamine, potassium acetate, sodium tert-butoxide, potassium tert-butoxide and sodium n-butoxide, the inorganic bases include but are not limited to sodium hydride, potassium phosphate, sodium carbonate, potassium carbonate, Potassium acetate, cesium carbonate, sodium hydroxide and lithium hydroxide.

The above reaction is preferably carried out in a solvent. The solvents used include but are not limited to: acetic acid, methanol, ethanol, n-butanol, toluene, tetrahydrofuran, dichloromethane, petroleum ether, ethyl acetate, n-hexane, dimethyl sulfoxide, 1 , 4-dioxane, water or N, N-dimethylformamide.

detailed description

The following examples are used to further describe the present disclosure, but these examples do not limit the scope of the present disclosure.

Examples

The structure of the compound is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). The NMR shift (δ) is given in units of 10 ^-6 (ppm). NMR was measured with Bruker AVANCE-400 NMR instrument, the solvent was deuterated dimethyl sulfoxide (DMSO-d ₆ ), deuterated chloroform (CDCl ₃ ), deuterated methanol (CD ₃ OD), the internal standard was four Methylsilane (TMS).

The MS was measured with FINNIGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).

High performance liquid chromatography (HPLC) analysis uses Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489 high pressure liquid chromatograph.

Chiral HPLC analysis and determination using Agilent 1260 DAD high-performance liquid chromatography.

High performance liquid phase preparation uses Waters 2767, Waters 2767-SQ Detector2, Shimadzu LC-20AP and Gilson-281 preparative chromatographs.

For chiral preparation, Shimadzu LC-20AP preparative chromatograph was used.

CombiFlash rapid preparation instrument uses Combiflash Rf200 (TELEDYNE ISCO).

The thin-layer chromatography silica gel plate uses Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plate. The specification of the thin-layer chromatography (TLC) silica gel plate is 0.15mm ～ 0.2mm, and the specification of the thin-layer chromatography separation purification product is 0.4mm ~ 0.5mm.

Silica gel column chromatography generally uses Yantai Huanghai Silica 200-300 mesh silica gel as a carrier.

The average value of ₅₀ measured kinase inhibition rate and IC NovoStar using a microplate reader (BMG, Germany).

The known starting materials of the present disclosure can be synthesized using or following methods known in the art, or can be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Darryl Chemical companies.

There is no special description in the examples, and the reaction can be carried out under an argon atmosphere or a nitrogen atmosphere.

An argon atmosphere or a nitrogen atmosphere means that the reaction bottle is connected to an argon or nitrogen balloon with a volume of about 1 L.

The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon with a volume of about 1L.

The pressurized hydrogenation reaction uses Parr3916EKX type hydrogenation instrument and Qinglan QL-500 hydrogen generator or HC2-SS type hydrogenation instrument.

The hydrogenation reaction is usually evacuated, filled with hydrogen, and repeated 3 times.

The microwave reaction uses CEM Discover-S 908860 microwave reactor.

There is no special description in the examples, and the solution refers to an aqueous solution.

There is no special description in the examples, and the reaction temperature is room temperature, which is 20 ° C to 30 ° C.

The monitoring of the reaction progress in the examples uses thin layer chromatography (TLC), the developing agent used in the reaction, the eluent system of column chromatography used for purifying the compound, and the developing agent system of thin layer chromatography include: A: Dichloromethane / methanol system, B: n-hexane / ethyl acetate system, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of basic or acidic reagents such as triethylamine and acetic acid can also be added for adjustment.

Example 1

2-methoxyethyl (6-butoxy-1- (4-pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) Carbamate

first step

2-methoxyethyl (6-butoxy-1- (4-pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) Carbamate

1- (4- (pyrrolidin-1-ylmethyl) benzyl) -6-butoxy-1H-pyrazolo [3,4-d] pyrimidin-4-amine 1a (100 mg, 0.26 mmol, using It is prepared by the well-known method "WO2018095426 (A1)". It is added to 5mL of N, N-dimethylformamide, cooled in an ice bath to 0-5 ° C, stirred, and sodium hydride (50mg, 1.25mmol, 60%, dispersed in mineral oil), stirred at 0-5 ° C for 30 minutes, and 2-methoxyethyl chloroformate 1b (70 mg, 0.51 mmol, Sigma-Aldrich), 0-5 was added dropwise to the reaction solution React for 1 hour at ℃. To the reaction solution, 10 mL of ice water was added dropwise to quench the reaction, extraction with ethyl acetate (10 mL × 3), the organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), dried over anhydrous magnesium sulfate, and filtered The drying agent was removed, the filtrate was concentrated under reduced pressure, and the resulting residue was purified using high-performance liquid chromatography (Gilson-281, elution system: acetonitrile, water) to obtain the title product 1 (14 mg, yield: 11.1%).

MS m / z (ESI): 483.3 [M + 1]

¹ H NMR (400MHz, CDCl ₃ ) δ8.53 (s, 1H), 7.51-8.16 (br, 1H), 7.21-7.34 (m, 4H), 5.50 (s, 2H), 4.29-4.47 (m, 4H ), 3.66 (dd, J = 5.6, 3.5 Hz, 2H), 3.58 (s, 2H), 3.41 (s, 3H), 2.40-2.56 (m, 4H), 1.68-1.85 (m, 6H), 1.50 ( q, J = 7.5 Hz, 2H), 0.98 (t, J = 7.5 Hz, 3H).

Example 2

(Ethane-1,2-diylbis (oxy)) bis (ethane-2,1-diyl) bis ((6-butoxy-1- (4- (pyrrolidin-1-ylmethyl Group) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) carbamate)

first step

(Ethane-1,2-diylbis (oxy)) bis (ethane-2,1-diyl) bis (perfluorophenyl) bis (carbonate)

Add 2,3,4,5,6-pentafluorophenol 2a (500mg, 2.72mmol, Shaoyuan) to 10mL of dichloromethane, add N-methylmorpholine (450mg, 4.45mmol), cool in ice bath Stir to 0-5 ° C, and drop (ethane-1,2-diylbis (oxy)) bis (ethane-2,1-diyl) dichloroformate 2b (375mg) to the reaction solution dropwise , 1.36 mmol, Alfa Aesar), react at 0-5 ℃ for 1 hour. To the reaction solution was added dropwise 10 mL of ice water to quench the reaction, extraction with dichloromethane (10 mL × 3), the organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), dried over anhydrous magnesium sulfate, and filtered The drying agent was removed, concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title product 2c (500 mg, yield: 64.6%).

MS m / z (ESI): 588.1 [M + 18]

¹ H NMR (400 MHz, CDCl ₃ ) δ 4.47-4.52 (m, 4H), 3.82-3.86 (m, 4H), 3.73 (s, 4H).

Second step

(Ethane-1,2-diylbis (oxy)) bis (ethane-2,1-diyl) bis ((6-butoxy-1- (4- (pyrrolidin-1-ylmethyl (Yl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) carbamic acid tert-butyl ester)

Add 1a (100 mg, 0.26 mmol) to 5 mL of N, N-dimethylformamide, cool to 0-5 ° C in an ice bath, stir, and add sodium hydride (50 mg, 1.25 mmol, 60%, dispersed in batches) In mineral oil), stir at 0-5 ° C for 30 minutes, dropwise add 2c (80 mg, 0.14 mmol) to the reaction solution, and react at 0-5 ° C for 1 hour. To the reaction solution, 10 mL of ice water was added dropwise to quench the reaction, extraction with ethyl acetate (10 mL × 3), the organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), dried over anhydrous magnesium sulfate, and filtered The drying agent was removed, the filtrate was concentrated under reduced pressure, and the resulting residue was purified using high-performance liquid chromatography (Gilson-281, elution system: acetonitrile, water) to obtain the title product 2 (25 mg, yield: 10.0%).

MS m / z (ESI): 964.3 [M + 1]

¹ H NMR (400 MHz, CDCl ₃ ) δ 13.06 (br, 2H), 8.54 (s, 2H), 7.31-7.42 (m, 8H), 5.52 (s, 4H), 4.32-4.47 (m, 8H), 4.18 (s, 4H), 3.73-3.83 (m, 4H), 3.70 (s, 4H), 3.63 (m, 4H), 2.80 (m, 4H), 2.08-2.36 (m, 8H), 1.73-1.82 ( m, 4H), 1.43-1.55 (m, 4H), 0.97 (t, J = 7.5 Hz, 6H).

Example 3

(6-Butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) carbamic acid decyl ester

first step

Decyl (perfluorophenyl) carbonate

Add 2a (500 mg, 2.72 mmol) to 10 mL of dichloromethane, add N-methylmorpholine (900 mg, 8.90 mmol), cool in an ice bath to reduce the temperature to 0-5 ° C, stir, and add chloroformic acid to the reaction solution dropwise Decyl ester 3a (900 mg, 4.08 mmol, TCI) was reacted at 0-5 ° C for 1 hour. To the reaction solution was added dropwise 10 mL of ice water to quench the reaction, extraction with dichloromethane (10 mL × 3), the organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), dried over anhydrous magnesium sulfate, and filtered The drying agent was removed, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title product 3b (500 mg, yield: 50.0%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 4.33 (t, J = 6.8 Hz, 2H), 1.71-1.85 (m, 2H), 1.20-1.49 (m, 14H), 0.82-0.99 (m, 3H).

Second step

(6-Butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) carbamic acid decyl ester

Add 1a (100 mg, 0.26 mmol) to 5 mL of N, N-dimethylformamide, cool to 0-5 ° C in an ice bath, stir, and add sodium hydride (50 mg, 1.25 mmol, 60% dispersed in minerals) in portions In oil), stirred at 0-5 ° C for 30 minutes, 3b (150mg, 0.41mmol) was added dropwise to the reaction solution, and reacted at 0-5 ° C for 1 hour. 5mL of ice water was added dropwise to the reaction solution to quench the reaction, extracted with ethyl acetate (10mL × 3), the organic phases were combined, washed with water (10mL), saturated sodium chloride solution (10mL), dried over anhydrous magnesium sulfate, and filtered The drying agent was removed, the filtrate was concentrated under reduced pressure, and the resulting residue was purified using high-performance liquid chromatography (Gilson-281, elution system: acetonitrile, water) to obtain the title product 3 (40 mg, yield: 27.2%).

MS m / z (ESI): 565.3 [M + 1]

¹ H NMR (400 MHz, CDCl ₃ ) δ 13.14 (br, 1H), 8.57 (s, 1H), 7.32-7.46 (m, 4H), 5.54 (s, 2H), 4.40 (t, J = 6.8Hz, 2H), 4.25 (t, J = 6.8Hz, 2H), 4.19 (s, 2H), 3.64 (br, 2H), 2.80 (br, 2H), 1.94-2.06 (m, 4H), 1.76-1.86 (m , 2H), 1.64-1.76 (m, 2H), 1.47-1.56 (m, 2H), 1.18-1.43 (m, 14H), 0.99 (t, J = 7.4Hz, 3H), 0.81-0.91 (m, 3H ).

Example 4

(6-Butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) carbamic acid octadecyl ester

first step

Octadecyl (perfluorophenyl) carbonate

2a (900mg, 4.89mmol) was added to 20mL of dichloromethane, N-methylmorpholine (1.50g, 14.83mmol) was added, the temperature was cooled to 0-5 ° C in an ice bath, stirring, and chlorine was added dropwise to the reaction solution A DMF solution of stearyl formate 4a (1.50 g, 4.51 mmol, prepared by the well-known method "US6686462 (B2)") was reacted at 0-5 ° C for 1 hour. To the reaction solution was added dropwise 10 mL of ice water to quench the reaction, extraction with dichloromethane (10 mL × 3), the organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), dried over anhydrous magnesium sulfate, and filtered The drying agent was removed, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title product 4b (1.30 g, yield: 60.0%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 4.33 (t, J = 6.8 Hz, 2H), 1.71-1.79 (m, 2H), 1.25-1.49 (m, 30H), 0.80-0.91 (m, 3H).

Second step

carbamate 4 (6-butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) carbamate ester

Add 1a (100 mg, 0.26 mmol) to 5 mL of N, N-dimethylformamide, cool to 0-5 ° C in an ice bath, stir, and add sodium hydride (50 mg, 1.25 mmol, 60% dispersed in minerals) in portions (In oil), stirred at 0-5 ° C for 30 minutes, 4b (150mg, 0.31mmol, dissolved in 5mL N, N-dimethylformamide) was added dropwise to the reaction solution, and reacted at 0-5 ° C for 1 hour. To the reaction solution, 10 mL of ice water was added dropwise to quench the reaction, extraction with ethyl acetate (10 mL × 3), the organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), dried over anhydrous magnesium sulfate, and filtered The drying agent was removed, the filtrate was concentrated under reduced pressure, and the resulting residue was purified using high-performance liquid chromatography (Gilson-281, elution system: acetonitrile, water) to obtain product 4 (20 mg, yield: 11.4%).

MS Jack / ES (ESI): 677.5 [M + 1]

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.56 (s, 1H), 7.66 (s, 1H), 7.31-7.39 (m, 4H), 5.52 (s, 2H), 4.40 (t, J = 6.8Hz, 2H), 4.25 (t, J = 6.8Hz, 2H), 3.79 (br, 2H), 2.74 (br, 2H), 1.89 (br, 4H), 1.79-1.83 (m, 2H), 1.68-1.71 (m , 2H), 1.41-1.61 (m, 2H), 1.22-1.36 (m, 32H), 0.99 (t, J = 7.4Hz, 3H), 0.86-0.97 (m, 3H).

Example 5

first step

Triphosgene (2.0g, 6.74mmol) was added to 20mL of tetrahydrofuran, cooled in an ice bath to 0-5 ° C, pyridine (6.5g, 82.1748mmol) was added, and polyethylene glycol 550 monomethyl ether was added dropwise to the reaction solution 5a (average molecular weight 550, 2.0g, 3.6mmol), react at 0-5 ° C for 3 hours. The reaction solution was concentrated under reduced pressure, the residue was dissolved with dichloromethane 20mL, the organic phase was washed with 1mol / L dilute hydrochloric acid solution (10mL × 3), saturated sodium chloride solution (10mL) and water (10mL), anhydrous sulfuric acid After drying with magnesium, the desiccant was removed by filtration, and the filtrate was concentrated under reduced pressure to obtain the title product 5b (2.50 g, yield: 68.7%). The product was directly used in the next reaction without purification.

Second step

Add 1a (100 mg, 0.26 mmol) to 5 mL of N, N-dimethylformamide, cool to 0-5 ° C in an ice bath, stir, and add sodium hydride (50 mg, 1.25 mmol, 60% dispersed in minerals) in portions In oil), stirred at 0-5 ° C for 30 minutes, and 5b (150mg, 0.31mmol, dissolved in 5mL N, N-dimethylformamide) was added dropwise to the reaction solution, warmed to room temperature to continue the reaction for 1 hour, to the reaction solution 10 mL of ice water was added dropwise to quench the reaction, extracted with ethyl acetate (10 mL × 3), the organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), dried over anhydrous magnesium sulfate, and filtered to remove the drying agent The filtrate was concentrated under reduced pressure, and the resulting residue was purified using high-performance liquid chromatography (Gilson-281, elution system: acetonitrile, water) to obtain product 5 (20 mg, yield: 17.3%).

MS / m / z (ESI): 833.4 [M + 1] (n = 9, n is the average)

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.54 (s, 1H), 7.29-7.33 (m, 4H), 5.51 (s, 2H), 4.36-4.42 (m, 4H), 3.81 (br, 2H), 3.62-3.79 (m, 2H), 3.60-3.75 (m, 34H), 3.52-3.58 (m, 2H), 3.38 (s, 3H), 2.75 (br, 4H), 1.89-2.06 (m, 6H), 1.80-1.86 (m, 4H), 1.77-1.81 (m, 2H), 1.48-1.54 (m, 2H), 0.99 (t, J = 7.4Hz, 3H).

Example 6

2- (decyloxy) ethyl (6-butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine-4 -Group) carbamate

first step

2- (decyloxy) ethyl chloroformate

Triphosgene (1.5g, 5.05mmol) was added to 20mL of dichloromethane, cooled to 0-5 ° C in an ice bath, N, N-diisopropylethylamine (1.0g, 7.74mmol) was added to the reaction solution Add 6a (0.5g, 2.47mmol, prepared by the well-known method "Chemistry-A European Journal, 2002, vol. 8, 4,900-909", dissolved in 10mL dichloromethane), reaction at 0-5 ℃ 2 hour. The reaction solution was concentrated under reduced pressure, and the residue was dissolved in 20 mL of dichloromethane. The organic phase was washed with 1 mol / L dilute hydrochloric acid solution (10 mL × 3), saturated sodium chloride solution (10 mL), and water (10 mL) successively, without water. It was dried over magnesium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to obtain the title product 6b (650 mg, yield: 99.3%), which was directly used in the next reaction without purification.

Second step

2- (decyloxy) ethyl (perfluorophenyl) carbonate

Add 2a (500mg, 2.72mmol) to 10mL of dichloromethane, add N-methylmorpholine (900mg, 8.90mmol), cool in an ice bath to reduce the temperature to 0-5 ° C, stir, and add 6b ( 650mg, 2.45mmol), react at 0-5 ° C for 1 hour. To the reaction solution was added dropwise 10 mL of ice water to quench the reaction, extracted with dichloromethane (10 mL × 3), the organic phases were combined, washed sequentially with water (10 mL), saturated sodium chloride solution (10 mL), and dried over anhydrous magnesium sulfate, The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title product 6c (430 mg, yield: 42.5%).

¹ H NMR (400 MHz, CDCl ₃ ) δ 4.42-4.52 (m, 2H), 3.74 (dd, J = 5.3, 4.0 Hz, 2H), 3.50 (t, J = 6.7 Hz, 2H), 1.60 (d, J = 7.3Hz, 2H), 1.15-1.44 (m, 14H), 0.83-0.95 (m, 3H).

third step

2- (decyloxy) ethyl (6-butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine-4 -Group) carbamate

Add 1a (100 mg, 0.26 mmol) to 5 mL of N, N-dimethylformamide, cool to 0-5 ° C in an ice bath, stir, and add sodium hydride (50 mg, 1.25 mmol, 60% dispersed in minerals) in portions (In oil), stirred at 0-5 ° C for 30 minutes, 6c (300 mg, 0.72 mmol) was added dropwise to the reaction solution, and reacted at 0-5 ° C for 1 hour. To the reaction solution, 5 mL of ice water was added dropwise to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), and dried over anhydrous magnesium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was purified using high-performance liquid chromatography (Gilson-281, elution system: acetonitrile, water) to obtain the title product 6 (35 mg, yield: 24.0%).

MS m / z (ESI): 609.1 [M + 1]

¹ H NMR (400 MHz, CDCl ₃ ) δ 8.54 (s, 1H), 7.84 (br. 1H), 7.24-7.31 (m, 4H), 5.50 (s, 2H), 4.32-4.48 (m, 4H), 3.64-3.74 (m, 2H), 3.58 (s, 2H), 3.48 (t, J = 6.8Hz, 2H), 2.44-2.54 (m, 4H), 1.72-1.85 (m, 6H), 1.43-1.64 ( m, 4H), 1.23-1.34 (m, 14H), 0.98 (t, J = 7.4 Hz, 3H), 0.83-0.92 (m, 3H).

Example 7

PEG5K-CM-Glycine-N-6-butoxy-1- (4-((pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine-4- amine

first step

(2-((6-butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) amino) -2-oxoethyl) t-butyl carbamate 7b

Add 1a (700 mg, 1.82 mmol) to 15 mL of N, N-dimethylformamide, cool it to 0-5 ° C in an ice bath, stir, and add sodium hydride (200 mg, 5.0 mmol, 60% dispersed in minerals) in portions In oil), stirred at 0-5 ° C for 30 minutes, 7a (1.7g, 4.98mmol prepared by a well-known method "J. Org. Chem. 2010, 75, 3, 564-569") was added dropwise to the reaction liquid, and the temperature was raised The reaction was continued to room temperature for 2 hours. To the reaction solution, 5 mL of ice water was added dropwise to quench the reaction, and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with water (10 mL), saturated sodium chloride solution (10 mL), and dried over anhydrous magnesium sulfate. The drying agent was removed by filtration, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by silica gel column chromatography with eluent system A to obtain the title product 7b (470 mg, yield: 52.1%).

MS m / z (ESI): 538.3 [M + 1]

Second step

2-amino-N- (6-butoxy-1- (4- (pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidin-4-yl) Acetamide

7b (470 mg, 0.87 mmol) was dissolved in 10 mL of dichloromethane, 5 mL of trifluoroacetic acid was added dropwise to the solution, stirred at room temperature for 1 hour, the reaction solution was spin-dried, diluted with 10 mL of ethyl acetate, and the organic phase was saturated with hydrogen carbonate The sodium solution (10 mL × 3) and water (10 mL) were washed, dried over anhydrous magnesium sulfate, filtered to remove the drying agent, and the filtrate was concentrated under reduced pressure to obtain the title product 7c (360 mg, yield: 94.1%).

MS m / z (ESI): 438.3 [M + 1]

third step

PEG5K-CM-Glycine-N-6-butoxy-1- (4-((pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine-4- amine

Add 7c (90mg, 0.21mmol) and one-arm-PEG5K-SCM (M-SCM-5K, methoxypolyethylene glycol succinimide acetate) (150mg, Mw: 5000, Beijing Kaijian) to To 5mL of N, N-dimethylformamide, N, N-diisopropylethylamine (45mg, 0.35mmol) was added and reacted at room temperature for 16 hours. Spin off most of the solvent under reduced pressure. The crude product is mixed with water and acetonitrile (V: V = 10: 1). The biological dialysis membrane is filtered three times. The filtrate is lyophilized to obtain the final product 7 (75.0 mg, loading rate 8.0 (w / w)%)

¹ H NMR (400 MHz, D ₂ O) δ 8.34 (s, 1.05H), 7.32-7.49 (m, 4.19H), 5.54 (s, 2.18H), 4.45 (br.s., 2.10H), 4.32 -4.35 (m, 3.09H), 4.16 (s, 1.30H), 3.42-3.83 (m, 454H), 3.33 (s, 3.22H), 3.11 (br.s., 4.32H), 2.02-2.15 (m , 4.16H), 1.78-1.95 (m, 2.24H), 1.68-1.77 (m, 2.39H), 1.32-1.43 (m, 2.42H), 0.90 (t, 2.99H).

Example 8

PEG10K-CM-Glycine-N-6-butoxy-1- (4-((pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine-4- amine

first step

Add 7c (90mg, 0.21mmol) and single-arm-PEG10K-SCM (M-SCM-10K, methoxypolyethylene glycol succinimide acetate) (150mg, Mw: 10000, Beijing Kaijian) to To 5mL of N, N-dimethylformamide, N, N-diisopropylethylamine (45mg, 0.35mmol) was added and reacted at room temperature for 16 hours. Spin off most of the solvent under reduced pressure. The crude product is mixed with water and acetonitrile (V: V = 10: 1). The biodialysis membrane is filtered three times. The filter cake solution is lyophilized to obtain the final product 8 (85.0 mg, loading rate 3.7 ( w / w)%)

¹ H NMR (400 MHz, D ₂ O) δ 8.34 (s, 0.97H), 7.25-7.55 (m, 3.59H), 5.53 (s, 2.00H), 4.44 (br.s., 2.09H), 4.32 -4.35 (m, 2.27H), 4.15 (s, 1.36H), 3.45-3.83 (m, 909H), 3.33 (s, 3.29H), 3.06-3.19 (m, 4.05H), 2.02-2.15 (m, 2.10H), 1.80-1.95 (m, 2.11H), 1.68-1.78 (m, 2.17H), 1.35-1.45 (m, 2.08H), 1.29 (d, 2.23H), 0.89 (t, 2.98H).

Example 9

Four-arm-PEG5K-CM-glycine-N-6-butoxy-1- (4-((pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine -4-amine

first step

7c (90mg, 0.21mmol) and 4-arm-PEG5K-SCM (F-SCM-5K, four-arm methoxypolyethylene glycol succinimide acetate) (150mg, Mw: 5000, Beijing Kaijian ) Was added to 5mL of N, N-dimethylformamide, N, N-diisopropylethylamine (45mg, 0.35mmol) was added and reacted at room temperature for 16 hours. Spin off most of the solvent under reduced pressure. The crude product is mixed with water and acetonitrile (V: V = 10: 1). The biodialysis membrane is filtered three times. The filter cake solution is lyophilized to obtain the final product 9 (50 mg, loading rate 27.7 (w / w)%).

¹ H NMR (400MHz, D ₂ O) δ 8.29 (br.s., 3.64H), 7.25-7.45 (m, 12.28H), 5.38 (br.s., 7.09H), 4.25-4.42 (m, 12.37H), 4.11-4.23 (m, 13.26H), 3.25-3.82 (m, 484H), 3.10-3.25 (m, 13.63H), 1.85-2.05 (m, 12.63H), 1.64-1.80 (m, 7.11) H), 1.34-1.48 (m, 7.17H), 0.86-0.95 (m, 9.58H).

Example 10

Four-arm-PEG20K-CM-glycine-N-6-butoxy-1- (4-((pyrrolidin-1-ylmethyl) benzyl) -1H-pyrazolo [3,4-d] pyrimidine -4-amine

first step

7c (82mg, 0.18mmol) and 4-arm-PEG20K-SCM (F-SCM-20K, four-arm methoxypolyethylene glycol succinimide acetate) (220mg, Mw: 20000, Beijing Kaijian ) Was added to 4mL of N, N-dimethylformamide, N, N-diisopropylethylamine (24mg, 0.18mmol) was added and reacted at room temperature for 16 hours. Spin off most of the solvent under reduced pressure, and add 30 mL of anhydrous ether to the reaction solution. A large amount of white solid is precipitated. The white solid is filtered, recrystallized three times with isopropanol, and filtered to dryness to obtain the final product of white solid 10 (62 mg, The load ratio is 7.8 (w / w)%).

¹ H NMR (400 MHz, D ₂ O) δ 8.25 (s., 3.59H), 7.27-7.34 (m, 14.95H), 5.44 (s., 8.18H), 4.33-4.37 (m, 10.74H), 4.20-4.22 (m, 14.64H), 3.25-3.82 (m, 1818H), 2.82-3.01 (m, 13.6H), 1.95-2.05 (m, 12.46H), 1.73-1.88 (m, 13.23H), 1.62 -1.66 (m, 11.24H), 1.29-1.34 (m, 11.48H), 0.79-0.82 (m, 11.97H).

Test case:

Biological evaluation

Test Example 1. Determination of the agonistic activity of the compounds of the present disclosure on human TLR7

The activation of hTLR7 protein expressed by HEK-Blue ^TM hTLR7 stable transfectant cells by the disclosed compounds is determined by the following experimental method:

1. Experimental materials and instruments

1.DMEM (Gibco, 10564-029),

2. Fetal bovine serum (GIBCO, 10099),

3. Penicillin (Gibco, 15140-122),

4. Trypan blue solution (Sigma, T8154-100ML),

5.Flexstation 3 multi-functional microplate reader (Molecμlar Devices),

6.HEK-Blue ^TM HTLR7 cell line (InvivoGen, hkb-hTLR7),

7. HEK-Blue detection reagent (InvivoGen, hb-det3).

2. Experimental procedure

Configure HEK-Blue test medium, take a bag of HEK-Blue test dry powder, add 50mL of endotoxin-free water to dissolve it, put it in a 37 ° C incubator, and sterile filter after 10 minutes. The compound was first formulated into a 20 mM stock solution; then it was diluted with pure DMSO to a maximum concentration of ⁶ × 10 ⁶ nM, and was diluted by a 3-fold gradient for a total of 10 points.

The compound prepared above was first diluted 20-fold with the medium, and then 20 μl of the diluted compound was added to each well. Take HEK-Blue ^TM hTLR7 cells, remove the supernatant first, then add 2-5mL of pre-warmed PBS, put it into the incubator for 1-2 minutes, gently pipette the cells, trypan blue staining and counting. Resuspend the cells in HEK-Blue detection medium to adjust the concentration to 2.2 × 10 ⁵ cells / mL, add 180 μL of cells to the 96-well cell culture plate to which 20 μL of drug has been added, and incubate at 37 ° C for 6-16 h.

Active molecule 1 was tested as a positive compound in the same way as above.

Microplate reader reading, the wavelength is 620nm. Corresponding OD values obtained by Graphpad Prism calculated EC ₅₀ value of the drug.

Compounds of the present disclosure human TLR7 activating effect can be determined by the above test, the measured EC ₅₀ values are shown in Table 1.

Table 1 EC _{50 of} compounds of the present disclosure against human-derived TLR7.

Conclusion: The compound of the present disclosure has no activation effect on TLR7, and the corresponding active molecule has a significant activation effect on TLR7, indicating that the compound of the present disclosure is a prodrug compound.

Test Example 2. Plasma metabolism evaluation test of the disclosed compounds

1. Summary

Using rat, mouse and human plasma as substrates, the compounds of the examples of the present disclosure and the above three different plasmas were determined using Thermo QE high-resolution liquid mass spectrometry, and were incubated at 37 ° C for 2 hours, respectively. The release of active molecule 1. The metabolic behavior of the compounds of the present disclosure in the plasma of three different species of rats, mice and humans was studied to evaluate their metabolic characteristics.

2. Test plan

2.1 Test drug

Examples Compound 1, Compound 2, Compound 3, Compound 4, Compound 5, Compound 6, Compound 7, Compound 8, Compound 9 and Compound 10

2.2 Test plasma

SD rat plasma and C57 mouse plasma are prepared in our laboratory. Human plasma is donated by volunteers of our company with the consent of the company. SD rats and C57 mice were purchased from Shanghai Jiejie Experimental Animal Co., Ltd. or Beijing Viton Lihua Experimental Animal Technology Co., Ltd.

2.3 Drug preparation

Weigh a certain amount of drug, add DMSO to make a 20mM stock solution, remove a certain volume of the stock solution and dilute it with a 50% acetonitrile solution to a 2mM working solution.

2.4 Sample incubation

Take 2 μL of 2 mM working solution and add to 198 μL of plasma of different species to make the final concentration of the compound 10 μM. The samples were incubated in a 37 ° C water bath for 120 minutes.

3. Sample processing and testing

After the incubation, 800 μL of acetonitrile was added, centrifuged at 12,000 rpm for 10 min, and the supernatant was evaporated in a vacuum evaporator. Reconstitute with 200 μL of 50% acetonitrile. The complex solution was injected and analyzed on a QE high-resolution liquid-mass spectrometer with a sample volume of 10 μL.

4. Results

The metabolism of the compounds of the present disclosure in the plasma of three species is as follows:

Conclusion: It can be seen from the above table that Example 5 has the largest proportion of active molecules 1 metabolized in human plasma, Example 3 has the least proportion of active molecules 1 metabolized in human plasma, and other embodiments release the proportion of active molecules. Between the two.

The main difference in the structure of the embodiments of the present disclosure is that the tail chain is different. For example, the tail chain of Example 5 is-(CH ₂ CH ₂ O) _n -CH ₃ , and the tail chain of Example 3 is -C ₁₀ alkyl. The difference is only the structure of the tail chain, the proportion of the active molecules released in the plasma of different species varies greatly, which fully shows that by changing the structure of the tail chain -R ⁰ -R in the general formula (I) of the present disclosure, the prodrug can be adjusted The rate at which compounds release active molecules.

Claims (30)

A compound represented by general formula (I):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: Y is selected from -C (O) O-, -C (O) NH-, -C (O)-, -C (O) (CH ₂ ) _s- , -C (O) NH (CH ₂ ) _s- Or -C (O) (CR ^a R ^b ) _s NHC (O) CH _2- ; R ^a and R ^{b are the} same or different, and are each independently selected from a hydrogen atom, halogen, alkyl, haloalkyl, hydroxy, or hydroxyalkyl; R ^{0 is} selected from-(CH ₂ CH ₂ O) _n- , alkylene,-(CH ₂ CH ₂ O) _n -alkylene-,-(OCH ₂ CH ₂ ) _n -O- or-(OCH ₂ CH ₂ ) _n -O-alkylene-; R is selected from _{-CH 3, -CH 2 CH 2 -} , - CH 2 -, - CH- , or C; Ring A is selected from cycloalkyl, heterocyclyl, aryl and heteroaryl; G is CH or N; X ¹ is alkylene or S (O) _m , wherein the alkylene is optionally selected from halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro , Cycloalkyl and heterocyclic groups are substituted by one or more substituents; L ^{1 is} selected from -NR ^4- , -O-, -S-, -C (O)-, -C (O) -OR ⁴ , -S (O) _m- , -N (R ⁴ ) C (O )-, -C (O) N (R ⁴ )-, -N (R ⁴ ) S (O) _2- , -S (O) ₂ N (R ⁴ )-and covalent bonds; R ^{1 is} selected from alkyl, alkoxy, haloalkyl, alkenyl, alkynyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, alkenyl, alkynyl, cycloalkane The group, heterocyclic group, aryl group and heteroaryl group are each independently selected from alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, One of heterocyclic group, aryl group, heteroaryl group, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ Or substituted by multiple substituents; R ^{2 is the} same or different and is independently selected from hydrogen atom, halogen, alkyl, alkoxy, haloalkyl, hydroxyl, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aromatic Group and heteroaryl group, wherein the alkyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group are each independently selected from the group consisting of alkyl group, alkoxy group, halogen, halogenated alkyl group, hydroxyl group, hydroxyl group Alkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ One or more substituents in R ⁷ and -C (O) NR ⁶ R ⁷ are substituted; L ² is an alkylene group or a covalent bond, wherein the alkylene group is optionally selected from alkyl, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cyclic Alkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ Substituted by one or more substituents in R ^{3 is} selected from haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclyl, aryl, heteroaryl, -OR ⁵ , -C (O) R ⁵ , -S (O) _m R ⁵ , -NR ⁶ R ⁷ and -C (O) NR ⁶ R ⁷ , wherein the cycloalkyl, heterocyclic, aryl and heteroaryl groups are each independently selected from Group, alkoxy, halogen, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl, heterocyclic, aryl, heteroaryl, -OR ⁸ , -C (O) R ^8. -S (O) _m R ⁸ , -NR ⁹ R ¹⁰ and -C (O) NR ⁹ R ¹⁰ are substituted by one or more substituents; R ^{4 is} selected from hydrogen atom, alkyl group, haloalkyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group; R ^{5 is} selected from hydrogen atom, alkyl group, haloalkyl group, amino group, hydroxyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group; R ⁶ and R ^{7 are the} same or different, and are each independently selected from a hydrogen atom, alkyl group, haloalkyl group, cycloalkyl group, heterocyclic group, aryl group, heteroaryl group, -C (O) R ⁸ , -S ( O) _m R ⁸ and -C (O) NR ⁹ R ¹⁰ ; wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl groups are each independently selected from alkyl, alkoxy Substituted by one or more substituents in the group, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; Alternatively, the R ⁶ and R ⁷ together with the connected nitrogen atom form a heterocyclic group, wherein the heterocyclic group may contain 1 to 2 same or different options in addition to 1 nitrogen atom. Heteroatoms from N, O and S, and the heterocyclic group is optionally selected from alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, hetero Substituted by one or more substituents in the cyclic group, aryl group and heteroaryl group; R ^{8 is} selected from hydrogen atom, alkyl group, haloalkyl group, amino group, hydroxyl group, cycloalkyl group, heterocyclic group, aryl group and heteroaryl group; R ⁹ and R ^{10 are the} same or different, and are each independently selected from a hydrogen atom, an alkyl group, a halogenated alkyl group, an amino group, a hydroxyl group, a cycloalkyl group, a heterocyclic group, an aryl group, and a heteroaryl group; t is 0, 1, 2, 3 or 4; s is an integer from 1 to 6; n is an integer from 0 to 460; q is 1, 2, 3, or 4; and m is 0, 1, or 2. The compound represented by the general formula (I) according to claim 1, wherein the ring A is phenyl or pyridyl. The compound represented by the general formula (I) according to claim 1 or 2, wherein X ¹ is an alkylene group; L ^{1 is} selected from -O-, -NR ^4- , -C (O)-and -C (O) N (R ⁴ )-, R ⁴ is a hydrogen atom or an alkyl group. The compound represented by general formula (I) according to any one of claims 1 to 3, which is a compound represented by general formula (II) or general formula (III):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: Y, R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in claim 1. The compound represented by the general formula (I) according to claim 1, which is a compound represented by the general formula (IV):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: Y is selected from -C (O) O- or ^{-C (O) (CR a R} b) s NHC (O) CH 2 -; R a and R ^{b are} identical or different and are each independently selected from hydrogen atoms or alkoxy Base, s is an integer from 1 to 3; R ^{0 is} selected from-(CH ₂ CH ₂ O) _n- , alkylene,-(CH ₂ CH ₂ O) _n -alkylene-,-(OCH ₂ CH ₂ ) _n -O- or-(OCH ₂ CH ₂ ) _n -O-alkylene-; q, R, R ¹ , L ² and R ^{3 are} as defined in claim 1; n is the average of 1 to 460. The compound represented by the general formula (I) according to any one of claims 1 to 5, wherein the R ³ is a heterocyclic group, and the heterocyclic group is optionally selected from alkyl, alkoxy Substituted by one or more substituents in the group, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclic, aryl and heteroaryl; R ³ is preferably -NR ⁶ R ⁷ , R ⁶ and R ⁷ together with the connected nitrogen atom form a heterocyclic group, wherein the heterocyclic group contains 1 nitrogen atom, and optionally contains 1 to 2 identical or different Heteroatoms of N, O and S, and the heterocyclic group is optionally selected from alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxyl, hydroxyalkyl, cycloalkyl, heterocyclic One or more substituents in the group, aryl group and heteroaryl group are substituted. The compound represented by general formula (I) according to any one of claims 1 to 6, which is a compound represented by general formula (IIA) or general formula (IIIA):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: R ⁶ and R ⁷ together with the connected nitrogen atom form a heterocyclic group, wherein the heterocyclic group contains 1 nitrogen atom, and optionally contains 1 or 2 same or different selected from N, O And S heteroatoms, and the heterocyclic group is optionally selected from alkyl, alkoxy, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aromatic Substituted by one or more substituents in the group and heteroaryl; R ⁰ , G, L ² , R ¹ , R ² and t are as defined in claim 1. The compound represented by general formula (I) according to any one of claims 1 to 7, wherein said G is N. The compound represented by general formula (I) according to any one of claims 1 to 8, wherein L ² is an alkylene group; R ^{2 is the} same or different, and each is independently a hydrogen atom or a halogen. The compound represented by the general formula (I) according to claim 1, which is a compound represented by the general formula (V):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: Y, R ¹ , R ⁰ , q, R are as defined in the general formula (IV). The compound represented by general formula (I) according to any one of claims 1 to 10, wherein R ¹ is an alkyl group, wherein the alkyl group is optionally substituted with one or more alkoxy groups. The compound represented by general formula (I) according to any one of claims 1 to 11, which is a compound represented by general formula (IIB) or general formula (IIIB):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: Y and R ^{0 are} as defined in claim 1. The compound represented by general formula (I) according to any one of claims 1 to 6 or 12, wherein said Y is -C (O) O- or -C (O)-. The compound represented by general formula (I) according to any one of claims 1 to 13, wherein said R ⁰ is-(CH ₂ CH ₂ O) _n- (CH ₂ ) _p- , n is 0 Integer to 20, p is an integer from 0 to 18, n and p are not 0 at the same time; preferably-(CH ₂ CH ₂ O) _n -,-(CH ₂ ) _p -or -CH ₂ CH ₂ O- ( CH ₂ ) _p- , n is 1, or an integer of 8 to 14, and p is an integer of 8 to 18. The compound represented by general formula (I) according to any one of claims 1 to 13, wherein said Y is -C (O) O-, R ^{0 is} selected from-(CH ₂ CH ₂ O) _n -, Alkylene or-(CH ₂ CH ₂ O) _n -alkylene-, n is as defined in claim 1. The compound represented by general formula (I) according to any one of claims 1 to 12, wherein Y is -C (O) (CR ^a R ^b ) _s NHC (O) CH _2- , R ⁰ is _{- (OCH 2 CH 2) n} -O- or _{- (OCH 2 CH 2) n} -O- alkylene ^{-, R a, R b,} n, s are as defined in claim 1. The compound represented by the general formula (I) according to claim 1, which is a compound represented by the general formula (VI):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: p is an integer of 0 to 18; R ¹ , q, R or n are as defined in the general formula (IV). The compound represented by the general formula (I) according to claim 1, which is a compound represented by the general formula (VII):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: R ¹ , q, R or n are as defined in the general formula (IV). The compound represented by the general formula (IIB) according to claim 12, which is a compound represented by the general formula (IIC):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: n is an integer from 0 to 20; p is an integer from 0 to 18; and n and p are not 0 at the same time. The compound represented by general formula (I) according to any one of claims 1 to 19, wherein said n is an average of 8 to 460, preferably an average of 10-460. The compound represented by the general formula (IIIB) according to claim 12, which is a compound represented by the general formula (IIIC):

Or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof or a pharmaceutically acceptable salt thereof, among them: n is an integer from 1 to 10. The compound represented by general formula (I) according to any one of claims 1 to 21, which is selected from:

n is an average of 8 to 500, preferably an average of 10-460. A method for preparing a compound represented by the general formula (II) according to claim 4, the method comprising:

The compound of formula (IA) and the compound of formula (IB) are reacted to obtain the compound of formula (II); among them: W is a leaving group, preferably halogen or

Y is -C (O) O- or -C (O)-; R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in claim 4. A method for preparing a compound represented by general formula (III) according to claim 4, the method comprising:

2 molecules of the compound of the general formula (IA) react with 1 molecule of the compound of the general formula (IC) to obtain the compound of the general formula (III); among them: W is a leaving group, preferably halogen or

Y is -C (O) O- or -C (O)-; R ⁰ , G, L ² , R ¹ to R ³ and t are as defined in claim 4. A method for preparing a compound represented by general formula (III) according to claim 4, the method comprising:

The compound of general formula (VI-A) and the compound of (VI-B) react under basic conditions to obtain the compound of general formula (VI); Among them, R ¹ , R and p are as defined in the general formula (VI). A pharmaceutical composition containing a therapeutically effective amount of a compound according to any one of claims 1 to 22 or a tautomer, a racemate, a racemate, and an enantiomer Isomers, diastereomers, or mixtures thereof or pharmaceutically acceptable salts thereof, and one or more pharmaceutically acceptable carriers, diluents or excipients. The compound according to any one of claims 1 to 22 or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof , Or a pharmaceutically acceptable salt thereof, or the use of the pharmaceutical composition according to claim 26 in the preparation of a medicament for agonizing TLR7. The compound according to any one of claims 1 to 22 or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof , Or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 26, for use in the preparation of a medicament for the treatment or prevention of a disease mediated by TLR7. The compound according to any one of claims 1 to 22 or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof , Or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 26 for use in the preparation of a medicament for treating or preventing tumors; wherein the tumors are preferably selected from melanoma, lung cancer, small cell lung cancer, Non-small cell lung cancer, hepatocellular carcinoma, basal cell carcinoma, renal cell carcinoma, myeloma, breast cancer, ovarian cancer, colon cancer, rectal cancer, prostate cancer, brain tumor, esophageal cancer, colorectal cancer, gastric cancer, liver cancer, pancreas Cancer, colorectal cancer, renal cancer, skin cancer, neuroblastoma, sarcoma, osteochondroma, osteosarcoma, seminoma, testicular tumor, uterine cancer, head and neck cancer, multiple myeloma, malignant lymphoma, true Polycythemia, leukemia, Hodgkin's disease, adrenal cortical tumors, thyroid tumors, ureteral tumors, bladder cancer, gallbladder cancer, cholangiocarcinoma, chorionic epithelial cancer, and pediatric tumors. The compound according to any one of claims 1 to 22 or a tautomer, a racemate, a racemate, an enantiomer, a diastereomer, or a mixture thereof , Or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 26 for use in the preparation of a medicament for treating infections caused by viruses selected from the group consisting of: dengue virus, yellow fever virus, sini Virus, Japanese encephalitis virus, tick-borne encephalitis virus, Kunjin virus, Murray valley encephalitis virus, Saint Louis encephalitis virus, Omsk hemorrhagic fever virus, bovine viral diarrhea virus, Zika virus, HIV HBV, HCV, HPV, RSV, SARS and influenza viruses.