CN111909204B - A tenofovir diphenylpropionate-based phosphoramidate compound and its pharmaceutical composition and application - Google Patents

Abstract

The invention discloses a tenofovir bisphenylpropionate-based phosphoramidate compound and a salt thereof, a preparation method and a pharmaceutical composition containing the compound. Tests have proved that the compound of the invention has the activity of inhibiting HBV virus replication, and at the same time, the compound has the advantages of higher activity and larger development coefficient than the current AIDS drug tenofovir disoproxil fumarate (TAF). Experiments also prove that the compound of the present invention also has the activity of inhibiting HIV-1 virus replication, and can be used for the development of AIDS medicine or hepatitis B medicine.

Description

A tenofovir diphenylpropionate-based phosphoramidate compound and its drug combination substance and use

technical field

The invention relates to the technical field of medicine, in particular to a tenofovir bisphenylpropionate-based phosphoramidate compound, a pharmaceutical composition containing the compound and use thereof.

Background technique

Hepatitis B, especially chronic hepatitis B, has become a global public health problem. According to statistics, more than 1 million people worldwide die from HBV infection and related diseases every year. But so far there is still a lack of ideal treatment methods and drugs for chronic hepatitis B. Therefore, how to effectively prevent and treat hepatitis B is a major problem that will be faced and needs to be solved in the 21st century.

At present, TAF (Valide) and ETV (Entecavir), the first-class drugs for the treatment of hepatitis B in the Chinese market, have gradually shown their respective shortcomings since they were launched.

In the 2018 American Society of Liver Diseases Prevention, Diagnosis and Treatment of Chronic Hepatitis B, entecavir (ETV), tenofovir disoproxil fumarate (TDF), and tenofovir alafenamide fumarate (TAF) were recommended For the best choice of drugs for the treatment of hepatitis B, each of the three has its own advantages and disadvantages:

Advantages of ETV: small side effects; Disadvantages: certain drug resistance, contraindicated for pregnant women;

Advantages of TDF: good antiviral effect, little drug resistance; Disadvantages: certain nephrotoxicity, long-term application will have certain side effects on bones and kidneys;

Advantages of TAF: It has the same antiviral effect as TDF, improves the safety parameters of kidney and bone laboratory, has very little drug resistance, and the dosage is only one-twelfth of that of Nofovir.

Vilide (TAF) is a new oral regimen for the treatment of hepatitis B. Although it cannot completely cure hepatitis B, its efficacy has been greatly improved, and it can well control the amount of hepatitis B virus. In terms of clinical effect, TAF and TDF have the same clinical effect in the treatment of hepatitis B, but the dosage of TAF is smaller, the toxicity is lower, and it will play a greater role in drug resistance and combination therapy. However, since TAF will shed a molecule of phenol after entering the human body, the TAF compound preparation has gradually shown toxic and side effects that cannot be ignored: lactic acidosis, severe hepatomegaly and fatty liver.

Therefore, it is of great significance to develop new analogues of TAF, maintain small changes in structure, maintain the excellent antiviral activity of TAF, and have better stability in plasma and oral bioavailability.

To sum up, although Gilead’s TDF and TAF compound preparations still dominate the global market for AIDS/Hepatitis B drugs, our research on TNF-based AIDS/Hepatitis B drugs is still of great significance. It is still of great value to further improve its bioavailability in the human body, increase its activity, reduce its toxic and side effects and give full play to its efficacy in the treatment of hepatitis B and AIDS.

Contents of the invention

In the process of prodrug research on TAF analogs, it was unexpectedly found that after the benzene ring chain of TAF was replaced by an amino NHR chain, the activity increased, and the biological activity selection coefficient SI also increased, which made the use of research samples Compared with the dose, TAF can be further reduced, that is, the toxicity of TAF can be further reduced, while maintaining the activity of TAF, and the drug concentration in tissue cells, especially liver cells, is significantly increased compared with TAF. Therefore, the tenofovir bisphenylpropionate-based phosphoramidate compound of the present invention can significantly improve the therapeutic effect on hepatitis B and AIDS, and greatly reduce the nephrotoxicity and bone toxicity caused by TDF or TAF. Accordingly, the following inventions have been proposed.

A tenofovir bisphenylpropionate-based phosphoramidate compound, comprising:

a) Compound (Ia), the structure of which is shown below,

Wherein, R ₁ =R ₂ =C ₁ -C ₁₂ alkyl; or

b) A pharmaceutically acceptable salt prepared from the compound (Ia) as described in a) and an acid.

Further, when the R ₁ and R ₂ groups are selected from isopropyl, ethyl, isobutyl, neopentyl, n-butyl, cyclohexyl, methyl or tert-butyl, the structural formulas of the compounds are as follows Structural formula:

Further, the pharmaceutically acceptable salt is oxalate, malonate, succinate, fumarate, aspartate, citrate, salicylate, hydrochloride, sulfate , phosphate, the range of molar ratio of acid and compound (Ia) is (0.5～1):1.

Further, when the R ₁ and R ₂ groups are selected from isopropyl, ethyl, isobutyl, neopentyl, n-butyl, cyclohexyl, methyl or tert-butyl, the fumarate The structural formula is the following structural formula respectively:

Further, the fumarate is prepared from fumaric acid and the compound at a molar ratio of 0.5:1.

On the other hand, the present invention also provides a pharmaceutical composition, comprising a therapeutically effective dose of tenofovir bisphenylpropionate phosphoramidate compound, and at least one pharmaceutically acceptable carrier or excipient.

Further, the pharmaceutical composition also comprises a therapeutically effective amount of an additional therapeutic agent selected from compounds that inhibit HIV protease, HIV non-nucleoside inhibitors of reverse transcriptase, HIV nucleoside inhibitors of reverse transcriptase, HIV nucleotide inhibitors of reverse transcriptase, HIV integrase inhibitors, gp41 inhibitors, CXCR4 inhibitors, gp120 inhibitors, CCR5 inhibitors, capsid polymerization inhibitors, non-catalytic site HIV integrase site inhibitors at least one.

Further, the dosage form of the pharmaceutical composition is tablet or capsule.

The tenofovir bisphenylpropionate-based phosphoramidate compound of the present invention can be applied to the preparation of medicines for preventing or treating viral diseases. A particular viral disease is HIV-1 infection or HBV infection or HIV-1 and HBV co-infection.

It has been determined that the compound of the present invention has the excellent properties required to become a drug for the treatment of AIDS/hepatitis B, specifically as follows:

In an in vitro anti-hepatitis B virus activity screening, the IC of compound _COP371371 is 6 times that of TAF (positive control), and the selection coefficient SI of biological activity is _4.0 times of TAF (positive control); the IC of compound COP374374 is TAF (positive control). ) of 6 times, the biological activity selection coefficient SI is 1.5 times of TAF (positive control). It shows that the compound of the present invention has higher activity of inhibiting virus replication than a class of anti-hepatitis B drug TAF, has a large selection coefficient of biological activity, and is expected to become a drug for treating HBV infection. The compound of the invention integrates various good properties such as high activity, low toxicity and high bioavailability, and has the prospect of becoming a new generation of medicine for treating AIDS or hepatitis B.

Detailed ways

The following examples can enable those skilled in the art to understand the present invention more fully, but do not limit the present invention in any way. The structures of all compounds were determined by ¹ H NMR or MS.

The raw materials used in the present invention: tenofovir, L-alanine ester hydrochloride and fumaric acid are all purchased from the market.

Example 1

9-[(R)-2-[[Di[[(S)-(1-n-butoxycarbonyl-2-phenyl)ethyl]amino]phosphinyl]methoxy]propyl]adenine Preparation of (COP374374).

Take commercially available tenofovir (COP00) 7.2g (25mmol), add in the 250ml flask, then add thionyl chloride 50g, heat up to 55°C, after 10 minutes, continue to heat up to 70°C and stir for 1 hour , distilled thionyl chloride under reduced pressure, added 100ml of dry acetonitrile after the distillation was completed and refluxed at 85°C for 10min, evaporated the acetonitrile to dryness under reduced pressure, cooled down to room temperature, then added 50ml of dry dichloromethane, and continued to stir for 0.5h Add 19.4g (75mmol, 3eq) of L-phenylalanine n-butyl hydrochloride (AH374), stir for another 10min, then add about 20ml of triethylamine dropwise, after the dropwise addition is complete, continue stirring for 0.5h.

After suction filtration, the filtrate was evaporated to dryness, and then 100ml of ethyl acetate was added. It was washed twice with 100 g of 5% potassium bicarbonate aqueous solution and twice with 100 g of 20% sodium chloride aqueous solution, dried over anhydrous sodium sulfate, suction filtered after drying, and the filtrate was evaporated to dryness. Ethyl acetate column chromatography yielded about 8 g of COP374374 (purity 98.2%).

¹ H NMR (400MHz, CDCl ₃ ) δ, (ppm): 0.85-0.98 (6H, dt, 2×CH ₃ ), 1.02-1.15 (3H, d, CH ₃ ), 1.25-1.41 (4H, m, 2 ×CH ₂ ), 1.45-1.67(4H,m,2×CH ₂ ),2.73-2.98(5H,m,OCH and 2×CH ₂ ),3.01-3.54(4H,m,2×NH and OCH ₂ P ),3.70-4.12(6H,m,2×NCH and 2×COOCH ₂ ),4.13-4.36(2H,m,NCH ₂ ),6.27(2H,s,NH ₂ ),7.00-7.38(10H,m, hydrogen on two benzene rings), 7.88 (1H, s, H on the purine ring), 8.32 (1H, s, H on the purine ring).

ESI-MS: [M+H]+694.7

Example 2

9-[(R)-2-[[Di[[(S)-(1-n-butoxycarbonyl-2-phenyl)ethyl]amino]phosphinyl]methoxy]propyl]adenine Preparation of Fumarate (FCOP374374).

An equivalent amount of 9-[(R)-2-[[two[[(S)-(1-n-butoxycarbonyl-2-phenyl)ethyl]amino]

Phosphinyl] methoxyl group] propyl group] adenine (COP374374) and fumaric acid were dissolved in hot acetonitrile, stirred at reflux for 2 hours, cooled and crystallized at room temperature, filtered out the precipitated solid and washed with acetonitrile to obtain a white solid: 9-[(R)-2-[[Di[[(S)-(1-n-butoxycarbonyl-2-phenyl)ethyl]amino]phosphinyl]methoxy]propyl]adenine Fumarate (FCOP374374).

Example 3

In Vitro Determination of Anti-HBV Viral Activity of Compounds of the Present Invention

1. In vitro cell model: HepG 2.2.15 cells

2. Dot blot method to determine the anti-hepatitis B virus activity of the compound

2. Put 1 HepG 2.2.15 cells (4×10 ⁴ cells/well) into a 96-well plate and culture overnight at 37°C with 5%.

2.2 On the second day, dilute the compound and add different concentrations of the compound to the culture wells. The final concentration of DMSO in the culture medium was 1%. 1 μM entecavir (ETV) was used as 100% Inhibition control; 1% DMSO was used as 0% Inhibition control.

2.3 On the fifth day, replace with fresh culture medium containing the compound.

2.4 On the eighth day and the ninth day, the culture medium in the culture well was removed, and the cells were harvested for dot hybridization.

3. Results: See Table 1.

Table 1: Cytotoxicity of compounds, extracellular anti-HBV activity (HepG2.2.15)

4. Experimental conclusion:

4.1 The positive compound TAF has the expected inhibitory effect on the replication of hepatitis B virus in HepG 2.2.15 cells, which proves that the experiment is effective and credible.

4.2 Anti-hepatitis B virus activity. In the dot hybridization experiment, the anti-hepatitis B virus activity of the two tested compounds in the above table was 6 times that of TAF (positive control). And the development coefficient SI is higher than the positive control TAF inhibition rate.

4.3 This fully shows that the compound of the present invention has higher activity in inhibiting virus replication than the current anti-hepatitis B drug TAF, and is expected to become a new generation of drugs for the treatment of HBV infection.

Example 4

Preparation formula and process

The pharmaceutical composition of the present invention can be made into tablet or capsule according to the preparation method of general oral pharmaceutical preparation, and the tablet or capsule unit content of the compound of the present invention of 5mg dosage is as follows:

Table II

name Proportion/mg Compounds of the invention 2.50 lactose 1.65 starch 0.66 microcrystalline cellulose 0.08 Carboxymethyl Starch Sodium 0.08 Magnesium stearate 0.03

.

Claims (4)

1. A tenofovir disoproxil phosphoramidate compound or a pharmaceutically acceptable salt thereof, wherein the structural formula of the tenofovir disoproxil phosphoramidate compound is selected from one of the following structural formulas:

；

the pharmaceutically acceptable salt is fumarate, and the structural formula of the fumarate is selected from one of the following structural formulas:

。

2. the tenofovir disoproxil phosphoramidate compound or a pharmaceutically acceptable salt thereof according to claim 1, wherein the molar ratio of fumaric acid of the fumarate to the tenofovir disoproxil phosphoramidate compound is in the range of (0.5-1): 1.

3. a pharmaceutical composition comprising a therapeutically effective amount of a tenofovir disoproxil phosphoramidate compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2, and at least one pharmaceutically acceptable carrier or excipient.

4. Use of a tenofovir dipivoxil phenylpropionate compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 2 in the manufacture of a medicament for the prevention or treatment of HBV infection viral diseases.