阅读说明：本技术 光学活性氯喹和羟氯喹及其类似物、其制备方法、组合物和用途 (Optically active chloroquine and hydroxychloroquine and analogs thereof, preparation method, composition and application thereof ) 是由 张 蒋晟 于 2020-07-17 设计创作，主要内容包括：本发明提供一种制备光学活性氯喹及其类似物的快速和简便的方法,将氯喹及其类似物的消旋体与酸性手性拆分试剂反应生成相应的盐,经分离纯化后得到光学纯的氯喹及其类似物的盐,再将其与碱反应得到高光学纯度的(R)-或(S)-氯喹及其类似物。此方法操作简便,成本较低,且对映体纯度可达99.9％ee,易于实现单一手性构型-氯喹及其类似物的工业化生产。本发明还提供一种(R)-或(S)-氯喹及其类似物、其药物组合物和用途,光学活性的氯喹及其类似物降低了神经系统和视网膜功能损伤,对冠状病毒、流感病毒等病毒造成的感染具有很好的抑制活性。(The invention provides a rapid and simple method for preparing optical activity chloroquine and analogues thereof, which comprises the steps of reacting racemates of the chloroquine and the analogues thereof with an acid chiral resolution reagent to generate corresponding salts, separating and purifying to obtain optically pure chloroquine and salts of the analogues thereof, and reacting the optically pure chloroquine and salts of the analogues thereof with alkali to obtain the (R) -or (S) -chloroquine and the analogues thereof with high optical purity. The method has simple operation and lower cost, the enantiomeric purity can reach 99.9% ee, and the industrial production of chloroquine and analogues thereof with single chiral configuration can be easily realized. The invention also provides (R) -or (S) -chloroquine and analogues thereof, pharmaceutical compositions and application thereof, wherein the optical active chloroquine and analogues thereof reduce the functional damage of nervous system and retina, and have good inhibitory activity on the infection caused by coronavirus, influenza virus and other viruses.)

1. A preparation method of a compound shown as a formula II or a formula III is characterized by comprising the following steps:

(a) reacting a compound shown as a formula I with a chiral resolution reagent in a solvent to generate salt, and filtering to obtain a salt of a compound (II) or a salt of a compound (III);

(b) recrystallizing the salt of compound (II) or the salt of compound (III) obtained in step (a) in a solvent;

(c) reacting the salt of compound (II) or compound (III) recrystallized in step (b) with a base in a solvent to obtain compound (II) or compound (III);

in the formula I, the formula II and the formula III,

R¹is halogen, amino, cyano, C_1-4Haloalkyl, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_3-6Cycloalkyl, or C_1-4A haloalkoxy group;

R²is unsubstituted or R^2-1Substituted C_1-6An alkyl group;

R³is hydrogen, or unsubstituted or R^3-1Substituted C_1-6An alkyl group;

R^2-1and R^3-1Independently selected from hydroxy, halogen, amino or cyano;

R⁴is hydrogen, unsubstituted or R^4-1Substituted C_1-6Alkyl or C_3-10A cycloalkyl group;

R^4-1is hydroxy, halogen, amino or cyano;

preferably, the solvent in step (a) is selected from any one of the following mixed solvents: methanol/ethyl acetate, ethanol/ethyl acetate, n-propanol/ethyl acetate, isopropanol/ethyl acetate, n-butanol/ethyl acetate, tert-butanol/ethyl acetate, tetrahydrofuran/ethyl acetate, acetone/ethyl acetate, acetonitrile/ethyl acetate, methanol/diethyl ether, ethanol/diethyl ether, n-propanol/diethyl ether, n-butanol/diethyl ether, or acetone/diethyl ether;

preferably, the chiral resolving agent described in step (a) is L-malic acid, D-tartaric acid, L-tartaric acid, D-mandelic acid, L-mandelic acid, (R) -1,1 '-binaphthol phosphate, (S) -1,1' -binaphthol phosphate, D-camphor-10-sulfonic acid, L-camphor-10-sulfonic acid, D- (+) -camphoric acid, L- (-) -camphoric acid, (+) -diacetyl-L-tartaric anhydride, (-) -di-p-toluoyl-L-tartaric acid, (+) -di-p-toluoyl-D-tartaric acid, or a mixture thereof, (+) -dibenzoyl-D-tartaric acid, (-) -dibenzoyl-L-tartaric acid, L-glutamic acid, L-glycine, D-aspartic acid, L-aspartic acid, D-pyroglutamic acid, L-pyroglutamic acid, or D- (-) -quinic acid;

preferably, in the preparation method, the ratio of the chiral resolution reagent in the step (a) to the compound shown in the formula I is 3: 1-5: 1;

preferably, the solvent in step (b) is selected from any one of the following mixed solvents: methanol/ethyl acetate, ethanol/ethyl acetate, n-propanol/ethyl acetate, isopropanol/ethyl acetate, n-butanol/ethyl acetate, tert-butanol/ethyl acetate, tetrahydrofuran/ethyl acetate, acetone/ethyl acetate, acetonitrile/ethyl acetate, methanol/diethyl ether, ethanol/diethyl ether, n-propanol/diethyl ether, n-butanol/diethyl ether, or acetone/diethyl ether; the recrystallization is carried out once, twice or more than three times;

preferably, the base in step (c) is one or a combination of two or more selected from sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, sodium propoxide, potassium propoxide, sodium isopropoxide, potassium N-butoxide, sodium t-butoxide, potassium t-butoxide, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, potassium bicarbonate, triethylamine, or N, N-diisopropylethylamine;

preferably, the solvent in step (c) is one or more mixed solvents selected from methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, acetonitrile, 1, 4-dioxane or tetrahydrofuran.

2. The method of claim 1, wherein when R is¹When the halogen is fluorine, chlorine, bromine or iodine;

and/or when R¹Is C_1-4When halogenated alkyl, said C_1-4Haloalkyl is-CF₃；

And/or when R¹Is C_1-4When alkyl, said C_1-4Alkyl is methyl, ethyl, propyl or isopropyl;

and/or when R¹Is C_1-4At alkoxy, said C_1-4Alkoxy is methoxy, ethoxy, propoxy or isopropoxy;

and/or when R¹Is C_3-6When there is a cycloalkyl group, said C_3-6Cycloalkyl is cyclopropyl;

and/or when R¹Is C_1-4When halogenated alkoxy, said C_1-4Haloalkoxy is-OCF₃；

And/or when R²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1Is one or more, when there are more than one R^2-1When R is said^2-1May be the same or different;

and/or when R²Is unsubstituted or R^2-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1Is one or more, when there are more than one R^3-1When R is said^3-1May be the same or different;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1Is one or more, when there are more than one R^4-1When R is said^4-1May be the same or different;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R⁴Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl being C_3-6A cycloalkyl group.

3. The method of claim 1, wherein when R is²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1The number of (a) is 1, 2 or 3;

and/or when R²Is unsubstituted or R^2-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

and/or when R²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1SubstitutionC of (A)_1-6Alkyl is

And/or when R³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1The number of (a) is 1, 2 or 3;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1Substituted C_1-6Alkyl is

And/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1The number of (a) is 1, 2 or 3;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1Substituted C_1-6Alkyl is

And/or when R⁴Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

4. The method of claim 1, wherein the compound of formula II or III is selected from the group consisting of:

or

5. A compound as shown in formula II or formula III, or pharmaceutically acceptable salt thereof,

wherein, in the formula II and the formula III,

R¹is halogen, amino, cyano, C_1-4Haloalkyl, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_3-6Cycloalkyl, or C_1-4A haloalkoxy group;

R²is unsubstituted or R^2-1Substituted C_1-6An alkyl group;

R³is hydrogen, or unsubstituted or R^3-1Substituted C_1-6An alkyl group;

R^2-1and R^3-1Independently selected from hydroxy, halogen, amino or cyano;

R⁴is hydrogen, unsubstituted or R^4-1Substituted C_1-6Alkyl, or C_3-10A cycloalkyl group;

R^4-1is hydroxy, halogen, amino or cyano.

6. A compound of formula II or III, or a pharmaceutically acceptable salt thereof, as claimed in claim 5, wherein when R is¹When the halogen is fluorine, chlorine, bromine or iodine;

and/or when R¹Is C_1-4When halogenated alkyl, said C_1-4Haloalkyl is-CF₃；

And/or when R¹Is C_1-4When alkyl, said C_1-4Alkyl is methyl, ethyl, propyl or isopropyl;

and/or when R¹Is C_1-4At alkoxy, said C_1-4Alkoxy is methoxy, ethoxy, propoxy or isopropoxy;

and/or when R¹Is C_3-6When there is a cycloalkyl group, said C_3-6Cycloalkyl is cyclopropyl;

and/or when R¹Is C_1-4When halogenated alkoxy, said C_1-4Haloalkoxy is-OCF₃；

And/or when R²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1Is one or more, when there are more than one R^2-1When R is said^2-1May be the same or different;

and/or when R²Is unsubstituted or R^2-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1Is one or more, when there are more than one R^3-1When R is said^3-1May be the same or different;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1Is one or more, when there are more than one R^4-1When R is said^4-1May be the same or different;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

and/or when R⁴Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl being C_3-6A cycloalkyl group.

7. A compound of formula II or III, or a pharmaceutically acceptable salt thereof, as claimed in claim 5, wherein when R is²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1The number of (a) is 1, 2 or 3;

and/or when R²Is unsubstituted or R^2-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

and/or when R²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1Substituted C_1-6Alkyl is

And/or when R³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1The number of (a) is 1, 2 or 3;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

and/or when R³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1Substituted C_1-6Alkyl is

And/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1The number of (a) is 1, 2 or 3;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

and/or when R⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1Substituted C_1-6Alkyl is

And/or when R⁴Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

8. The compound of formula II or formula III, or a pharmaceutically acceptable salt thereof, according to claim 5, wherein the compound of formula II or formula III is selected from the group consisting of:

or

9. A pharmaceutical composition comprising a compound of formula II or III, or a pharmaceutically acceptable salt thereof, as claimed in any one of claims 5-8, and a pharmaceutically acceptable adjuvant.

10. Use of a compound of formula II or formula III, or a pharmaceutically acceptable salt thereof, as claimed in any one of claims 5-8, or a pharmaceutical composition as claimed in claim 9, in the manufacture of a medicament for the prophylaxis or treatment of a viral infection;

preferably, the virus is middle east syndrome-associated coronavirus (MERS-CoV), severe acute respiratory syndrome-associated coronavirus (SARS-CoV), influenza A virus, influenza B virus, novel coronavirus pneumonia (COVID-19), rabies virus, poliovirus, AIDS virus, hepatitis A virus, hepatitis C virus, influenza A A H5N1 virus, chikungunya disease, dengue fever virus, Zika virus, lassa virus, Congo hemorrhagic fever virus, Ebola virus, hepatitis B virus or herpes simplex virus.

Technical Field

The invention belongs to the technical field of pharmaceutical chemistry, and particularly relates to optical pure (S) -chloroquine and an analogue thereof, an (R) -chloroquine and an analogue thereof, a chiral resolution method, a medicinal composition and application thereof.

Background

Chloroquine and hydroxychloroquine have been of great interest as antimalarial drugs for over the last 50 years. At present, the sulfate form of hydroxychloroquine, namely hydroxychloroquine sulfate, is one of the most commonly used drugs for the clinical treatment of rheumatic diseases, in particular systemic lupus erythematosus and rheumatoid arthritis. In the related mechanism research experiment, hydroxychloroquine has anti-inflammatory and immunoregulatory activity. Chloroquine has an ultra-long half-life, is easy to accumulate toxicity after continuous administration, and most patients have toxicity when the plasma concentration (valley concentration) is more than 0.8 mu g/mL. The heart toxicity and even death can be caused by more than or equal to 20mg/kg per day. Hydroxychloroquine is used as a common medicine for chronic diseases such as rheumatoid arthritis, systemic lupus erythematosus and the like, and irreversible retinal damage is easy to occur after long-term or high-dose taking.

Disclosure of Invention

The present inventors have conducted intensive studies to overcome the above-mentioned drawbacks and disadvantages of the prior art, and have unexpectedly found that (S) -configuration compounds or (R) -configuration compounds of (S) -chloroquine and hydroxychloroquine and (R) -chloroquine and their analogs are superior to racemic compounds in terms of pharmacological activity, toxicity, and the like. In view of the above, the present invention solves the above technical problems by the following technical solutions.

The invention provides a preparation method of a compound shown as a formula II or a formula III, which comprises the following steps:

(a) reacting a compound shown as a formula I with a chiral resolution reagent in a solvent to generate salt, and filtering to obtain a salt of a compound (II) or a salt of a compound (III);

(b) recrystallizing the salt of compound (II) or the salt of compound (III) obtained in step (a) in a solvent;

(c) reacting the salt of compound (II) or compound (III) recrystallized in step (b) with a base in a solvent to obtain compound (II) or compound (III);

in the formula I, the formula II and the formula III,

R¹is halogen, amino, cyano, C_1-4Haloalkyl, C_1-4Alkyl radical, C_1-4Alkoxy radical, C_3-6Cycloalkyl, or C_1-4A haloalkoxy group;

R²is unsubstituted or R^2-1Substituted C_1-6An alkyl group;

R³is hydrogen, or unsubstituted or R^3-1Substituted C_1-6An alkyl group;

R^2-1and R^3-1Independently selected from hydroxy, halogen, amino or cyano;

R⁴is hydrogen, unsubstituted or R^4-1Substituted C_1-6Alkyl or C_3-10A cycloalkyl group;

R^4-1is hydroxy, halogen, amino or cyano.

Preferably, in the preparation method, the solvent in the step (a) is selected from any one of the following mixed solvents: methanol/ethyl acetate, ethanol/ethyl acetate, n-propanol/ethyl acetate, isopropanol/ethyl acetate, n-butanol/ethyl acetate, tert-butanol/ethyl acetate, tetrahydrofuran/ethyl acetate, acetone/ethyl acetate, acetonitrile/ethyl acetate, methanol/diethyl ether, ethanol/diethyl ether, n-propanol/diethyl ether, n-butanol/diethyl ether, or acetone/diethyl ether.

Preferably, in the preparation method, the chiral resolving agent in the step (a) is selected from any one of the following: l-malic acid, D-tartaric acid, L-tartaric acid, D-mandelic acid, L-mandelic acid, (R) -1,1 '-binaphthol phosphate, (S) -1,1' -binaphthol phosphate, D-camphor-10-sulfonic acid, L-camphor-10-sulfonic acid, D- (+) -camphoric acid, L- (-) -camphoric acid, (+) -diacetyl-L-tartaric anhydride, (-) -di-p-toluoyl-L-tartaric acid, (+) -di-p-toluoyl-D-tartaric acid, (+) -dibenzoyl-D-tartaric acid, di-p-toluoyl-L-tartaric acid, di-n-toluoyl-L-tartaric acid, di-naphthoyl-D-tartaric acid, di-naphthoyl-L-tartaric acid, di-naphthoyl-tartaric acid, di-10-camphoric acid, di-acetyl-4-naphthoyl-tartaric acid, di-naphthoyl-4-naphthoyl-tartaric acid, di-4-naphthoyl-4, and a, (-) -dibenzoyl-L-tartaric acid, L-glutamic acid, L-glycine, D-aspartic acid, L-aspartic acid, D-pyroglutamic acid, L-pyroglutamic acid, or D- (-) -quinic acid.

Preferably, in the preparation method, the ratio of the chiral resolution reagent in the step (a) to the compound shown in the formula I is 3: 1-5: 1.

Preferably, in the preparation method, the solvent in the step (b) is selected from any one of the following mixed solvents: methanol/ethyl acetate, ethanol/ethyl acetate, n-propanol/ethyl acetate, isopropanol/ethyl acetate, n-butanol/ethyl acetate, tert-butanol/ethyl acetate, tetrahydrofuran/ethyl acetate, acetone/ethyl acetate, acetonitrile/ethyl acetate, methanol/diethyl ether, ethanol/diethyl ether, n-propanol/diethyl ether, n-butanol/diethyl ether, or acetone/diethyl ether. Preferably, the recrystallization may be performed once, twice or more.

Preferably, in the preparation method, the base in step (c) is one or a combination of two or more selected from sodium methoxide, potassium methoxide, sodium ethoxide, potassium ethoxide, sodium propoxide, potassium propoxide, sodium isopropoxide, potassium N-butoxide, sodium t-butoxide, potassium t-butoxide, sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, sodium bicarbonate, triethylamine, or N, N-diisopropylethylamine.

Preferably, in the preparation method, the solvent in step (c) is one or more mixed solvents selected from methanol, ethanol, n-propanol, isopropanol, n-butanol, tert-butanol, acetonitrile, 1, 4-dioxane or tetrahydrofuran.

Preferably, when R is¹When the halogen is fluorine, chlorine, bromine or iodine;

preferably, when R is¹When the halogen is fluorine, chlorine, bromine or iodine;

preferably, when R is¹Is C_1-4When halogenated alkyl, said C_1-4Haloalkyl is-CF₃；

Preferably, when R is¹Is C_1-4When alkyl, said C_1-4Alkyl is methyl, ethyl, propyl or isopropyl;

preferably, when R is¹Is C_1-4At alkoxy, said C_1-4Alkoxy is methoxy, ethoxy, propoxy or isopropoxy;

preferably, when R is¹Is C_3-6When there is a cycloalkyl group, said C_3-6Cycloalkyl is cyclopropyl;

preferably, when R is¹Is C_1-4When halogenated alkoxy, said C_1-4Haloalkoxy is-OCF₃；

Preferably, when R is²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1Is one or more, when there are more than one R^2-1When R is said^2-1May be the same or different;

preferably, when R is²Is unsubstituted or R^2-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

preferably, when R is³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1Is one or more, when there are more than one R^3-1When R is said^3-1May be the same or different;

preferably, when R is³Is unsubstituted or R^3-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

preferably, when R is⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1Is one or more, when there are more than one R^4-1When R is said^4-1May be the same or different;

preferably, when R is⁴Is not takenIs substituted or R^4-1Substituted C_1-6When alkyl, said C_1-6Alkyl is C_1-4An alkyl group;

preferably, when R is⁴Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl being C_3-6A cycloalkyl group.

Further preferably, when R is²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1The number of (a) is 1, 2 or 3;

preferably, when R is²Is unsubstituted or R^2-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

preferably, when R is²Is unsubstituted or R^2-1Substituted C_1-6When it is alkyl, said R^2-1Substituted C_1-6Alkyl isOr,

Preferably, when R is³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1The number of (a) is 1, 2 or 3;

preferably, when R is³Is unsubstituted or R^3-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

preferably, when R is³Is unsubstituted or R^3-1Substituted C_1-6When it is alkyl, said R^3-1Substituted C_1-6Alkyl isOr,

Preferably, when R is⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1The number of (a) is 1, 2 or 3;

preferably, when R is⁴Is unsubstituted or R^4-1Substituted C_1-6When alkyl, said C_1-6Alkyl is methyl, ethyl, propyl, butyl or isopropyl, preferably ethyl;

preferably, when R is⁴Is unsubstituted or R^4-1Substituted C_1-6When it is alkyl, said R^4-1Substituted C_1-6Alkyl isOr,

Preferably, when R is⁴Is C_3-10When there is a cycloalkyl group, said C_3-10Cycloalkyl is cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl.

Preferably, the compound represented by formula II or formula III is selected from the following compounds:

or

In another aspect, the present invention also provides a method for preparing a compound represented by formula II or formula III as described above, wherein the compound represented by formula II or formula III is not (R) -chloroquine, (S) -chloroquine, (R) -hydroxychloroquine, or (S) -hydroxychloroquine.

The invention also provides a compound shown as the formula II or the formula III or pharmaceutically acceptable salt thereof.

The invention also provides an application of the compound shown in the formula II or the formula III or the pharmaceutically acceptable salt thereof in preparing a medicament for preventing or treating virus infection.

Further, the virus is middle east syndrome related coronavirus (MERS-CoV), severe acute respiratory syndrome related coronavirus (SARS-CoV), influenza A virus, influenza B virus, novel coronavirus pneumonia (COVID-19), rabies virus, poliovirus, AIDS virus, hepatitis A virus, hepatitis C virus, influenza A A H5N1 virus, chikungunya disease, dengue fever virus, Zika virus, lassa virus, Congo hemorrhagic fever virus, Ebola virus, hepatitis B virus or herpes simplex virus.

The invention also provides a pharmaceutical composition which comprises the compound shown as the formula II, pharmaceutically acceptable salts thereof and pharmaceutic adjuvants.

Further, in the pharmaceutical composition, the compound shown in formula II or the pharmaceutically acceptable salt thereof is used in an amount of therapeutically effective amount.

The invention also provides application of the pharmaceutical composition in preparing a medicament for preventing or treating virus infection.

Further, the virus is middle east syndrome related coronavirus (MERS-CoV), severe acute respiratory syndrome related coronavirus (SARS-CoV), influenza A virus, influenza B virus, novel coronavirus pneumonia (COVID-19), rabies virus, poliovirus, AIDS virus, hepatitis A virus, hepatitis C virus, influenza A A H5N1 virus, chikungunya disease, dengue fever virus, Zika virus, lassa virus, Congo hemorrhagic fever virus, Ebola virus, hepatitis B virus or herpes simplex virus.

The invention also provides a pharmaceutical composition which comprises the compound shown as the formula III or pharmaceutically acceptable salt thereof and pharmaceutic adjuvant.

Further, in the pharmaceutical composition, the compound shown in the formula III or the pharmaceutically acceptable salt thereof is used in a therapeutically effective amount.

The invention also provides application of the pharmaceutical composition in preparing a medicament for preventing or treating virus infection.

Further, the virus is middle east syndrome related coronavirus (MERS-CoV), severe acute respiratory syndrome related coronavirus (SARS-CoV), influenza A virus, influenza B virus, novel coronavirus pneumonia (COVID-19), rabies virus, poliovirus, AIDS virus, hepatitis A virus, hepatitis C virus, influenza A A H5N1 virus, chikungunya disease, dengue fever virus, Zika virus, lassa virus, Congo hemorrhagic fever virus, Ebola virus, hepatitis B virus or herpes simplex virus.

The pharmaceutical excipients can be those widely used in the field of pharmaceutical production. The excipients are used primarily to provide a safe, stable and functional pharmaceutical composition and may also provide methods for dissolving the active ingredient at a desired rate or for promoting the effective absorption of the active ingredient after administration of the composition by a subject. The pharmaceutical excipients may be inert fillers or provide a function such as stabilizing the overall pH of the composition or preventing degradation of the active ingredients of the composition. The pharmaceutical excipients may include one or more of the following excipients: binders, suspending agents, emulsifiers, diluents, fillers, granulating agents, adhesives, disintegrating agents, lubricants, antiadherents, glidants, wetting agents, gelling agents, absorption delaying agents, dissolution inhibitors, reinforcing agents, adsorbents, buffering agents, chelating agents, preservatives, colorants, flavoring agents, sweeteners, and the like.

The pharmaceutical compositions of the present invention may be prepared according to the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, levigating, encapsulating, entrapping or lyophilizing processes.

The pharmaceutical compositions of the present invention may be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid formulations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implant, subcutaneous, intravenous, intraarterial, intramuscular) administration. The pharmaceutical compositions of the present invention may also be in a controlled release or delayed release dosage form (e.g., liposomes or microspheres). Examples of solid oral formulations include, but are not limited to, powders, capsules, caplets, soft capsules, and tablets. Examples of liquid formulations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs and solutions. Examples of topical formulations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serum formulations. Examples of formulations for parenteral administration include, but are not limited to, solutions for injection, dry preparations which can be dissolved or suspended in a pharmaceutically acceptable carrier, suspensions for injection, and emulsions for injection. Examples of other suitable formulations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic formulations; aerosol: such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories and lozenges.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of the present invention, prepared from the compounds of the present invention found to have particular substituents, with relatively nontoxic acids or bases. When compounds of the invention contain relatively acidic functional groups, base addition salts can be obtained by contacting free forms of such compounds with a sufficient amount of a base in neat solution or in a suitable inert solvent. Pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic ammonia or magnesium salts or similar salts. When compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting free forms of such compounds with a sufficient amount of an acid in neat solution or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include salts of inorganic acids including, for example, hydrochloric acid, hydrobromic acid, nitric acid, carbonic acid (forming a carbonate or bicarbonate), phosphoric acid ((forming a phosphate, monohydrogen phosphate, dihydrogen phosphate), sulfuric acid (forming a sulfate or bisulfate), hydroiodic acid, phosphorous acid, and the like, as well as salts of organic acids including similar acids such as acetic acid, propionic acid, isobutyric acid, maleic acid, malonic acid, benzoic acid, succinic acid, suberic acid, fumaric acid, lactic acid, mandelic acid, phthalic acid, benzenesulfonic acid, p-toluenesulfonic acid, citric acid, tartaric acid, and methanesulfonic acid, and salts of organic acids including amino acids such as arginine, and the like, as well as salts of organic acids such as glucuronic acid, certain specific compounds of the invention contain basic and acidic functional groups and thus can be converted to any base or acid addition salt. The free form of the compound is regenerated by contacting the salt with a base or acid and isolating the parent compound in a conventional manner. The free form of the compound differs from its various salt forms in certain physical properties, such as solubility in polar solvents.

The "pharmaceutically acceptable salts" of the present invention can be synthesized from the parent compound containing an acid or base by conventional chemical methods. In general, such salts are prepared by the following method: prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid, in water or an organic solvent or a mixture of the two. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred.

The invention has the beneficial effects that:

(1) the preparation method of the optical active chloroquine/hydroxychloroquine and the analogue thereof has the advantages of simple and convenient operation, high resolution yield, higher optical purity of the product, realization of industrial production and wide application prospect.

(2) Compared with racemes, the single-configuration chloroquine/hydroxychloroquine and the analogues thereof have better antiviral activity and smaller toxic and side effects.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

Example 1: preparation of L-malate salt of (R) -hydroxychloroquine

The racemate hydroxychloroquine (5g, 14.88mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), L-malic acid (6g, 44.64mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of L-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the racemate at 0 ℃. Heating to 5 ℃, reacting for 30min, dissolving most of the L-malic acid salt of (S) -hydroxychloroquine in the ethanol/ethyl acetate mixed solution until the reaction is complete, precipitating the L-malic acid salt of (R) -hydroxychloroquine, and filtering to obtain crude L-malic acid salt of (R) -hydroxychloroquine (5.2 g).

Example 2: recrystallization purification of L-malate of (R) -hydroxychloroquine

Weighing and suspending an L-malate crude product (5.2g) of (R) -hydroxychloroquine in 10mL of ethanol/ethyl acetate (v/v is 1:2), heating and refluxing to be completely dissolved, slowly cooling to room temperature, keeping the temperature, stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain an L-malate (4.3g) of (R) -hydroxychloroquine with the optical purity of 87.8 percent ee.. Then, the L-malate salt of (R) -hydroxychloroquine obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain an L-malate salt of (R) -hydroxychloroquine (3.8g) having an optical purity of ee. of 98.8%.

Example 3: preparation of (R) -hydroxychloroquine

Sodium methoxide (1g,20.24mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, and L-malate salt of (R) -hydroxychloroquine (3.8g, 5.06mmol) was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (R) -hydroxychloroquine (1.6g), wherein the purity is more than 99%, the chiral purity is more than 99.92%, and the yield is 64%.

Example 4: preparation of D-malate salt of (S) -hydroxychloroquine

The racemate hydroxychloroquine (5g, 14.88mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), D-malic acid (6g, 44.64mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of D-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the racemate at 0 ℃. Heating to 5 ℃, reacting for 30min, dissolving most of the salt of the D-malic acid of the (R) -hydroxychloroquine in the ethanol/ethyl acetate mixed solution until the reaction is complete, precipitating the D-malic acid salt of the (S) -hydroxychloroquine, and filtering to obtain a crude D-malic acid salt of the (S) -hydroxychloroquine (4.8 g).

Example 5: recrystallization purification of D-malate of (S) -hydroxychloroquine

Weighing a crude D-malate salt (4.8g) of (S) -hydroxychloroquine, suspending the crude D-malate salt in 10mL of ethanol/ethyl acetate (v/v is 1:2), heating and refluxing until the crude D-malate salt is completely dissolved, slowly cooling to room temperature, keeping the temperature, stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain a D-malate salt (4.3g) of (S) -hydroxychloroquine with the optical purity of 88.3 percent ee.. Then, the D-malate salt of (S) -hydroxychloroquine obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain a D-malate salt of (S) -hydroxychloroquine (3.6g) having an optical purity of ee. of 97.4%.

Example 6: preparation of (S) -hydroxychloroquine

Sodium methoxide (1g,19.17mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, and D-malate salt of (S) -hydroxychloroquine (3.6g, 4.79mmol) was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (S) -hydroxychloroquine (1.4g), wherein the purity is more than 99%, the chiral purity is more than 99.92%, and the yield is 56%.

Example 7: preparation of L-mandelate salts of (R) -hydroxychloroquine

The racemate hydroxychloroquine (5g, 14.88mol) was dissolved in methanol/ethyl acetate (v/v ═ 1:3, 40mL), L-mandelic acid (7.9g, 42.08mol) was dissolved in methanol/ethyl acetate (v/v ═ 1:3, 20mL), and a methanol/ethyl acetate solution of L-mandelic acid was slowly added dropwise to an ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 ℃, reacting for 30min, dissolving most of the L-mandelate of (S) -hydroxychloroquine in the mixed solution of methanol and ethyl acetate until the reaction is complete, separating out the L-mandelate of (R) -hydroxychloroquine, and performing suction filtration to obtain a crude L-mandelate product (5.2g) of (R) -hydroxychloroquine.

Example 8: recrystallization purification of L-mandelate salt of (R) -hydroxychloroquine

Weighing an L-mandelate crude product (5.2g) of (R) -hydroxychloroquine, suspending the L-mandelate crude product in 12mL of methanol/ethyl acetate (v/v is 1:3), heating and refluxing until the L-mandelate crude product is completely dissolved, slowly cooling to room temperature, keeping the temperature, stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain an L-mandelate (4.5g) of (R) -hydroxychloroquine with the optical purity of 88.3 percent ee.. Then, the L-mandelic acid salt of (R) -hydroxychloroquine obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain an L-mandelic acid salt of (R) -hydroxychloroquine (3.8g) having an optical purity of ee. of 97.2%.

Example 9: preparation of (R) -hydroxychloroquine

The L-mandelate salt of (R) -hydroxychloroquine (3.8g, 4.72mmol) was dissolved in 30mL of methanol, left at room temperature, potassium carbonate (2.7g,19.83mol) was added to the above solution in portions, stirred at room temperature for about 30min, filtered with suction, and concentrated under reduced pressure to remove methanol. Adding 20ml of ethyl acetate to dissolve, adding water for 5min, then extracting with ethyl acetate (20ml multiplied by 2), combining organic phases, washing with water (10ml multiplied by 2), drying with anhydrous sodium sulfate, filtering, concentrating, and drying in vacuum to obtain (R) -hydroxychloroquine (1.4g) with the purity of more than 99%, the chiral purity of more than 99.93%, and the yield of 56%.

Example 10: preparation of D-mandelate salts of (S) -hydroxychloroquine

The racemate hydroxychloroquine (5g, 14.88mol) was dissolved in methanol/ethyl acetate (v/v ═ 1:3, 40mL), D-mandelic acid (7.9g, 42.08mol) was dissolved in methanol/ethyl acetate (v/v ═ 1:3, 20mL), and a methanol/ethyl acetate solution of D-mandelic acid was slowly added dropwise to an ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 ℃, reacting for 30min, dissolving most of the D-mandelate of the (R) -hydroxychloroquine in the mixed solution of methanol and ethyl acetate until the reaction is complete, separating out the D-mandelate of the (S) -hydroxychloroquine, and performing suction filtration to obtain a crude D-mandelate product (5.4g) of the (S) -hydroxychloroquine.

Example 11: recrystallization purification of D-mandelate salt of (S) -hydroxychloroquine

Weighing a crude D-mandelate salt (5.4g) of (S) -hydroxychloroquine, suspending the crude D-mandelate salt in 12mL of methanol/ethyl acetate (v/v is 1:3), heating and refluxing until the crude D-mandelate salt is completely dissolved, slowly cooling to room temperature, keeping the temperature, stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain a D-mandelate salt (4.7g) of (S) -hydroxychloroquine with the optical purity of 87.2 percent ee.. Then, the D-mandelic acid of (S) -hydroxychloroquine obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain a D-mandelic acid salt of (S) -hydroxychloroquine (4.0g) having an optical purity of 98.3% ee..

Example 12: preparation of (S) -hydroxychloroquine

D-mandelate salt of (S) -hydroxychloroquine (4.0g, 4.96mmol) was dissolved in 30mL of methanol, and potassium carbonate (3g,22.32mol) was added to the above solution in portions at room temperature, followed by stirring at room temperature for about 30min, suction filtration, and concentration under reduced pressure to remove methanol. Adding 20ml of ethyl acetate to dissolve, adding water for 5min, then extracting with ethyl acetate (20ml multiplied by 2), combining organic phases, washing with water (10ml multiplied by 2), drying with anhydrous sodium sulfate, filtering, concentrating, and drying in vacuum to obtain (S) -hydroxychloroquine (1.4g) with the purity of more than 99%, the chiral purity of more than 99.95% and the yield of 56%.

Example 13: preparation of L-malate salt of (R) -chloroquine

The racemate chloroquine (5g, 15.67mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), L-malic acid (6.7g, 50.16mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of L-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the racemate at 0 ℃. Heating to 5 ℃, reacting for 30min, dissolving most of the L-malic acid salt of (S) -chloroquine in the ethanol/ethyl acetate mixed solution until the reaction is complete, precipitating the L-malic acid salt of (R) -chloroquine, and filtering to obtain crude L-malic acid salt of (R) -chloroquine (5.0 g).

Example 14: recrystallization purification of L-malate of (R) -chloroquine

Weighing and suspending an L-malate crude product (5.0g) of (R) -chloroquine in 10mL of ethanol/ethyl acetate (v/v is 1:2), heating and refluxing to be completely dissolved, slowly cooling to room temperature, keeping the temperature, stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain an L-malate (4.4g) of (R) -chloroquine with the optical purity of 87.8 percent ee.. Then, the L-malate salt of (R) -chloroquine obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain an L-malate salt of (R) -chloroquine (3.9g) having an optical purity of ee. of 98.8%.

Example 15: preparation of (R) -chloroquine

Sodium ethoxide (1.5g,21.64mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, L-malate salt of (R) -chloroquine (3.9g, 5.41mmol) was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (R) -chloroquine (1.7g), wherein the purity is more than 99%, the chiral purity is more than 99.92%, and the yield is 68%.

Example 16: preparation of D-malate salt of (S) -chloroquine

The racemate chloroquine (5g, 15.67mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), D-malic acid (6.7g, 50.16mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of D-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the racemate at 0 ℃. And (3) heating to 5 ℃, reacting for 30min, dissolving most of the salt of the D-malic acid of the (R) -chloroquine in the ethanol/ethyl acetate mixed solution until the reaction is complete, precipitating the D-malic acid salt of the (S) -chloroquine, and filtering to obtain a crude D-malic acid salt of the (S) -chloroquine (4.8 g).

Example 17: recrystallization purification of D-malate of (S) -chloroquine

Weighing a crude D-malate salt (4.8g) of (S) -chloroquine, suspending the crude D-malate salt in 10mL of ethanol/ethyl acetate (v/v is 1:2), heating and refluxing until the crude D-malate salt is completely dissolved, slowly cooling to room temperature, keeping the temperature, stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain a D-malate salt (4.3g) of (S) -chloroquine with the optical purity of 88.3 percent ee.. Then, the D-malate salt of (S) -chloroquine obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain a D-malate salt of (S) -chloroquine (3.6g) having an optical purity of ee. of 97.4%.

Example 18: preparation of (S) -chloroquine

Sodium ethoxide (1.36g,20.0mol) is added into 10ml of anhydrous methanol in batches, the mixture is stirred for about 10min at 0 ℃, then the system becomes clear, D-malate of (S) -chloroquine (3.6g, 5.0mmol) is slowly added, the mixture is stirred for 30min at room temperature, and the methanol is removed by concentration under reduced pressure. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (S) -hydroxychloroquine (1.4g), wherein the purity is more than 99%, the chiral purity is more than 99.92%, and the yield is 56%.

Example 19: preparation of (R) -N¹,N¹-diethyl-N⁴L-malic acid salt of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine

Racemic modification N¹,N¹-diethyl-N⁴- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (5g, 16.39mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), L-malic acid (7.7g, 57.37mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of L-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete and most of (S) -N¹,N¹-diethyl-N⁴A salt of L-malic acid of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine is dissolved in an ethanol/ethyl acetate mixed solution, and (R) -N¹,N¹-diethyl-N⁴Salting out L-malic acid salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine, and suction filtering to obtain (R) -N¹,N¹-diethyl-N⁴Crude L-malate salt of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (5.0 g).

Example 20: (R) -N¹,N¹-diethyl-N⁴Recrystallization purification of L-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine

Weighing (R) -N¹,N¹-diethyl-N⁴- (7-fluoro)Suspending a crude L-malate (5.0g) of quinoline-4-yl) pentyl-1, 4-diamine in 10mL of ethanol/ethyl acetate (v/v ═ 1:2), heating and refluxing to completely dissolve the crude L-malate, slowly cooling to room temperature, keeping the temperature, stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain (R) -N with the optical purity of 87.6 percent ee.¹,N¹-diethyl-N⁴L-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (4.4 g). Then, the (R) -N obtained above is recrystallized¹,N¹-diethyl-N⁴(R) -N with an optical purity of 97.9% ee. was obtained by primary recrystallization of L-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine¹,N¹-diethyl-N⁴L-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (3.9 g).

Example 21: (R) -N¹,N¹-diethyl-N⁴Preparation of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine

Sodium methoxide (1.3g,23.23mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear and (R) -N was slowly added¹,N¹-diethyl-N⁴L-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (3.9g, 5.53mmol), stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (R) -N¹,N¹-diethyl-N⁴- (7-Fluoroquinolin-4-yl) pentyl-1, 4-diamine (1.3g), purity>99% chiral purity>99.95% and a yield of 52%.

Example 22: preparation of (S) -N¹,N¹-diethyl-N⁴D-malic acid salt of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine

Racemic modification N¹,N¹-diethyl-N⁴- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (5g, 16.39mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL) and D-apple was addedFruit acid (7.7g, 57.37mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL) and the ethanol/ethyl acetate solution of D-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete and most of (R) -N¹,N¹-diethyl-N⁴The salt of D-malic acid of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine is dissolved in an ethanol/ethyl acetate mixed solution, and (S) -N¹,N¹-diethyl-N⁴Salting out D-malic acid salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine, and suction filtering to obtain (S) -N¹,N¹-diethyl-N⁴Crude D-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (4.7 g).

Example 23: (S) -N¹,N¹-diethyl-N⁴Recrystallization purification of D-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine

Weighing (S) -N¹,N¹-diethyl-N⁴Suspending the crude D-malate (4.7g) of the- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine in 10mL of ethanol/ethyl acetate (v/v ═ 1:4), heating and refluxing to completely dissolve the D-malate, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain (S) -N with the optical purity of 88.3% ee.¹,N¹-diethyl-N⁴D-malate salt of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (4.3 g). Then, the (S) -N obtained above is recrystallized¹,N¹-diethyl-N⁴(S) -N with an optical purity of 97.4% ee. was obtained by primary recrystallization of D-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine¹,N¹-diethyl-N⁴D-malate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (3.6 g).

Example 24: (S) -N¹,N¹-diethyl-N⁴Preparation of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine

Adding sodium ethoxide (1.36g,20.0mol) into 10ml of anhydrous methanol in batches, stirring at 0 deg.C for about 10min to clarify the system, and slowly adding (S) -N¹,N¹-diethyl-N⁴D-malate salt of (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (3.6g, 5.0mmol), stirred at room temperature for 30min, concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (S) -N¹,N¹-diethyl-N⁴- (7-Fluoroquinolin-4-yl) pentyl-1, 4-diamine (1.6g), purity>99% chiral purity>99.90% and yield 64%.

Example 25: preparation of (R) -N¹,N¹-diethyl-N⁴L-Camphorate salts of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine

Racemic modification N¹,N¹-diethyl-N⁴- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (5g, 14.16mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:4, 25mL), L-camphoric acid (11.6g, 58.07mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:4, 12mL), and the ethanol/ethyl acetate solution of L-camphoric acid was slowly added dropwise to the ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete and most of (S) -N¹,N¹-diethyl-N⁴The salt of L-camphoric acid of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine is dissolved in the ethanol/ethyl acetate mixed solution, and (R) -N¹,N¹-diethyl-N⁴Precipitating (R) -N with L-camphoric acid salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine, and vacuum filtering to obtain (R) -N¹,N¹-diethyl-N⁴Crude L-camphorate salt of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (5.8 g).

Example 26: (R) -N¹,N¹-diethyl-N⁴Recrystallization purification of L-Campholate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine

Weighing (R) -N¹,N¹-diethyl-N⁴Crude L-camphorate salt of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (5.8g) was suspended in 10mL ethanol/ethyl acetate (v/v ═ 1:4)Heating and refluxing to completely dissolve, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain (R) -N with the optical purity of 87.6 percent ee.¹,N¹-diethyl-N⁴L-Camphorate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (5.0 g). Then, the (R) -N obtained above is recrystallized¹,N¹-diethyl-N⁴(R) -N with an optical purity of 97.9% ee. was obtained by primary recrystallization of the L-camphorate salt of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine¹,N¹-diethyl-N⁴L-Camphorate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (4.7 g).

Example 27: (R) -N¹,N¹-diethyl-N⁴Preparation of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine

Sodium methoxide (1.1g,20.71mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear and (R) -N was slowly added¹,N¹-diethyl-N⁴L-Camphorate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (4.7g, 4.93mmol), stirred at room temperature for 30min, concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (R) -N¹,N¹-diethyl-N⁴- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (1.3g), purity>99% chiral purity>99.95% and a yield of 52%.

Example 28: preparation of (S) -N¹,N¹-diethyl-N⁴D-Camphorate salts of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine

Racemic modification N¹,N¹-diethyl-N⁴- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (5g, 14.16mol) in ethanol/ethyl acetate (v/v ═ 1:4, 25mL) and D-camphoric acid (11.6g, 58.07mol) in ethanol/ethyl acetateEthanol/ethyl acetate (v/v ═ 1:4, 12mL), D-camphoric acid in ethanol/ethyl acetate was slowly added dropwise to the ethanol/ethyl acetate solution of the racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete and most of (R) -N¹,N¹-diethyl-N⁴The salt of D-camphoric acid of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine is dissolved in the ethanol/ethyl acetate mixed solution, and (S) -N¹,N¹-diethyl-N⁴D-camphoric acid salt of- (7-trifluoromethyl quinoline-4-yl) amyl-1, 4-diamine is precipitated and filtered to obtain (S) -N¹,N¹-diethyl-N⁴Crude D-camphorate salt of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (4.7 g).

Example 29: (S) -N¹,N¹-diethyl-N⁴Recrystallization purification of D-Campholate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine

Weighing (S) -N¹,N¹-diethyl-N⁴Suspending the crude D-camphorate (4.7g) of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine in 10mL of ethanol/ethyl acetate (v/v ═ 1:2), heating and refluxing to completely dissolve, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain (S) -N with the optical purity of 88.3% ee.¹,N¹-diethyl-N⁴D-Camphorate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (4.5 g). Then, the (S) -N obtained above is recrystallized¹,N¹-diethyl-N⁴(S) -N with an optical purity of 97.4% ee. was obtained by primary recrystallization of D-camphorate of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine¹,N¹-diethyl-N⁴D-Camphorate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (3.7 g).

Example 30: (S) -N¹,N¹-diethyl-N⁴Preparation of (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine

Adding sodium ethoxide (1.1g,15.53mol) into 10ml of anhydrous methanol in batches, stirring at 0 deg.C for about 10min to clarify the system, and slowly adding (S) -N¹,N¹-diethyl-N⁴D-Camphorate salt of- (7-fluoroquinolin-4-yl) pentyl-1, 4-diamine (3.7g, 3.88mmol), stirred at room temperature for 30min, concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (S) -N¹,N¹-diethyl-N⁴- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (1.6g), purity>99% chiral purity>99.93%, yield 64%.

Example 31: preparation of L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol

The racemate 2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (5g, 13.55mol) was dissolved in acetonitrile/ethyl acetate (v/v ═ 1:4, 25mL), L-tartaric acid (7.1g, 47.42mol) was dissolved in acetonitrile/ethyl acetate (v/v ═ 1:4, 12mL), and the acetonitrile/ethyl acetate solution of L-tartaric acid was slowly added dropwise to the acetonitrile/ethyl acetate solution of the above racemate at 0 ℃. Heating to 10 ℃, reacting for 1h, dissolving most of the salt of the L-tartaric acid of the (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinoline-4-yl) amino) amyl) amino) ethyl-1-alcohol in the mixed solution of acetonitrile/ethyl acetate until the reaction is completed, and the L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol was precipitated and filtered to give a crude L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (5.2 g).

Example 32: recrystallization purification of L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol

Weighing a crude L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (5.2g) and suspending the crude L-tartrate salt in 10mL of acetonitrile/ethyl acetate (v/v ═ 1:4), heating and refluxing to be completely dissolved, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, further cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain an L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (4.8g) with the optical purity of 88.8 percent ee.. Then, the L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain an L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol having an optical purity of ee. of 98.2% (4.2 g).

Example 33: preparation of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol

Sodium methoxide (1.1g,20.52mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, and the L-tartrate salt of (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (4.2g, 5.13mmol) was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. 20ml of ethyl acetate is added, stirred for 1 hour at room temperature, filtered to remove salts, and the ethyl acetate mother liquor is concentrated under reduced pressure to obtain (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (1.3g), the purity is more than 99%, the chiral purity is more than 99.92%, and the yield is 52%.

Example 34: preparation of D-tartrate salt of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol

The racemate 2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (5g, 13.55mol) was dissolved in acetonitrile/ethyl acetate (v/v ═ 1:4, 25mL), D-tartaric acid (7.1g, 47.42mol) was dissolved in acetonitrile/ethyl acetate (v/v ═ 1:4, 12mL), and the acetonitrile/ethyl acetate solution of D-tartaric acid was slowly added dropwise to the acetonitrile/ethyl acetate solution of the above racemate at 0 ℃. Heating to 10 ℃, reacting for 1h, dissolving most of the salt of the D-tartaric acid of the (R) -2- (ethyl (4- ((7- (trifluoromethyl) quinoline-4-yl) amino) amyl) amino) ethyl-1-alcohol in the mixed solution of acetonitrile/ethyl acetate until the reaction is completed, the D-tartrate salt of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol was precipitated and filtered to give a crude D-tartrate salt of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (4.9 g).

Example 35: recrystallization purification of D-tartrate salt of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol

Weighing crude D-tartrate (4.9g) of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol, suspending the crude D-tartrate in 10mL of acetonitrile/ethyl acetate (v/v ═ 1:4), heating and refluxing to completely dissolve the crude D-tartrate, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, further cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain D-tartrate (4.8g) of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol with optical purity of 89.9 percent ee.. Then, the D-tartrate salt of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol obtained above was recrystallized once according to the above-mentioned recrystallization method to obtain a D-tartrate salt of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol having an optical purity of ee. of 98.6% (4.3 g).

Example 36: preparation of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol

Sodium methoxide (1.2g,21.53mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, and the D-tartrate salt of (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (4.3g, 5.25mmol) was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. 20ml of ethyl acetate is added, stirred for 1 hour at room temperature, filtered to remove salts, and the ethyl acetate mother liquor is concentrated under reduced pressure to obtain (S) -2- (ethyl (4- ((7- (trifluoromethyl) quinolin-4-yl) amino) pentyl) amino) ethyl-1-ol (1.4g), the purity is more than 99%, the chiral purity is more than 99.94%, and the yield is 56%.

Example 37: preparation of (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴1, 4-diamine L-methyl-pentyl-tartrate

Racemic modification N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Methyl pentyl-1, 4-diamine (5g, 15.02mol) was dissolved in methanol/ethyl acetate (v/v ═ 1:3, 30mL), L-tartaric acid (7.9g, 52.55mol) was dissolved in methanol/ethyl acetate (v/v ═ 1:3, 15mL), and the methanol/ethyl acetate solution of L-tartaric acid was slowly added dropwise to the methanol/ethyl acetate solution of the racemate at 0 ℃. Heating to 10 deg.C, reacting for 1h until the reaction is complete and most of (S) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴The salt of L-tartaric acid of (1, 4-methylpentyl-1, 4-diamine is dissolved in the mixed solution of acetonitrile/ethyl acetate, and (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Separating out L-tartrate of the (R) -N-methyl amyl-1, 4-diamine, and filtering to obtain (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Crude L-tartrate salt of (E) -methylpentyl-1, 4-diamine (5.0 g).

Example 38: (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Recrystallization purification of L-tartrate of (E) -methylpentyl-1, 4-diamine

Weighing (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Suspending the L-tartrate crude product (5.0g) of the (E) -methyl amyl-1, 4-diamine in 12mL of acetonitrile/ethyl acetate (v/v is 1:3), heating and refluxing until the L-tartrate crude product is completely dissolved, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain (R) -N with the optical purity of 87.9 percent ee.⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴L-tartrate salt of (E) -methylpentyl-1, 4-diamine (4.5 g). Then, it is recrystallized as described aboveThe method of (R) -N obtained as described above⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴The L-tartrate salt of (E) -methylpentyl-1, 4-diamine was recrystallized once to obtain N having an optical purity of 98.2% ee.⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴L-tartrate salt of (E) -methylpentyl-1, 4-diamine (4.2 g).

Example 39: (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Preparation of (E) -methylpentyl-1, 4-diamine

Sodium methoxide (1.2g,21.99mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear and (R) -N was slowly added¹,N¹-diethyl-N⁴L-tartrate salt of- (7-trifluoromethylquinolin-4-yl) pentyl-1, 4-diamine (4.2g, 5.36mmol), stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Methyl pentyl-1, 4-diamine (1.4g), purity>99% chiral purity>99.90% and yield 56%.

Example 40: preparation of (S) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴1, 4-diamine-methylpentyl-1, 4-diamine tartrate

Racemic modification N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Methyl pentyl-1, 4-diamine (5g, 15.02mol) was dissolved in tetrahydrofuran/ethyl acetate (v/v ═ 1:1, 40mL), D-tartaric acid (7.9g, 52.55mol) was dissolved in methanol/ethyl acetate (v/v ═ 1:1, 10mL), and the methanol/ethyl acetate solution of D-tartaric acid was slowly added dropwise to the tetrahydrofuran/ethyl acetate solution of the racemate at 0 ℃. Heating to 10 deg.C, reacting for 1h until the reaction is complete and most of (R) -N⁴- (7-chloroquinoline-4-Radical) -N¹,N¹-diethyl-N⁴The salt of D-tartaric acid of (S) -N is dissolved in a mixed solution of acetonitrile/ethyl acetate⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Separating out D-tartrate of the (R) -N-methyl amyl-1, 4-diamine, and filtering to obtain (R) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Crude D-tartrate salt of (4.8g) methylpentyl-1, 4-diamine.

Example 41: (S) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Recrystallization purification of D-tartrate of (E) -methylpentyl-1, 4-diamine

Weighing (S) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Suspending the crude D-tartrate (4.8g) of the (E) -methyl amyl-1, 4-diamine in 12mL tetrahydrofuran/ethyl acetate (v/v is 1:1), heating and refluxing to completely dissolve the D-tartrate, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, continuously cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain (S) -N with the optical purity of 88.5% ee.⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴D-tartrate salt of (E) -methylpentyl-1, 4-diamine (4.6 g). Then, the (S) -N obtained above is recrystallized⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴(S) -N with an optical purity of 98.3% ee. was obtained by one recrystallization of D-tartrate salt of (methylpentyl-1, 4-diamine)⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴D-tartrate salt of (E) -methylpentyl-1, 4-diamine (4.0 g).

Example 42: (S) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Preparation of (E) -methylpentyl-1, 4-diamine

Sodium methoxide (1.1g,20.95mol) was added in portions to 10ml of anhydrous methanol, and stirred at 0 ℃After about 10min, the system became clear and (S) -N was added slowly⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴D-tartrate salt of (E) -methylpentyl-1, 4-diamine (4.0g, 5.10mmol), stirred at room temperature for 30min, concentrated under reduced pressure to remove methanol. Adding 20ml ethyl acetate, stirring for 1 hour at room temperature, filtering to remove salt, and concentrating the ethyl acetate mother liquor under reduced pressure to obtain (S) -N⁴- (7-chloroquinolin-4-yl) -N¹,N¹-diethyl-N⁴Methyl pentyl-1, 4-diamine (1.2g), purity>99% chiral purity>99.92% and a yield of 48%.

Example 43: preparation of L-malate salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

The racemate 2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (5g, 14.33mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), L-malic acid (5.76g, 42.99mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of L-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete, dissolving most of (S) -2- ((4- ((7-chloroquinoline-4-yl) (methyl) amino) amyl) (ethyl) amino) ethyl-1-alcohol L-malic acid salt in ethanol/ethyl acetate mixed solution, and L-malic acid salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol was precipitated, and filtered under suction to give a crude L-malic acid salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (4.8 g).

Example 44: recrystallization purification of L-malate salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Weighing crude L-malate (4.8g) of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol, suspending the crude L-malate in 10mL of ethanol/ethyl acetate (v/v ═ 1:2), heating and refluxing to completely dissolve, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, further cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain L-malate (4.4g) of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol with optical purity of 87.8% ee.. Then, (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol obtained above was subjected to one-time recrystallization according to the above-mentioned recrystallization method to obtain an L-malate salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol having an optical purity of ee. (3.9 g).

Example 45: preparation of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Sodium methoxide (1.1g,20.77mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, and L-malate salt (3.9g, 5.19mmol) of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml of ethyl acetate, stirring at room temperature for 1 hour, carrying out suction filtration to remove salt, and concentrating an ethyl acetate mother liquor under reduced pressure to obtain (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (1.4g), wherein the purity is more than 99%, the chiral purity is more than 99.91%, and the yield is 56%.

Example 46: preparation of D-malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

The racemate 2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (5g, 14.33mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), D-malic acid (5.76g, 42.99mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of D-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete, dissolving most of (R) -2- ((4- ((7-chloroquinoline-4-yl) (methyl) amino) amyl) (ethyl) amino) ethyl-1-alcohol salt of D-malic acid in ethanol/ethyl acetate mixed solution, and D-malic acid salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol was precipitated, and filtered under suction to give a crude D-malate salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (4.6 g).

Example 47: recrystallization purification of D-malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Weighing crude D-malate (4.6g) of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol, suspending the crude D-malate in 10mL of ethanol/ethyl acetate (v/v ═ 1:2), heating and refluxing to completely dissolve, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, continuing cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain D-malate (4.2g) of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol with optical purity of 88.3% ee.. Then, (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol obtained above was subjected to one-time recrystallization according to the above-mentioned recrystallization method to obtain a D-malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol having an optical purity of ee. (3.7 g).

Example 48: preparation of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Sodium methoxide (1.1g,19.70mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, and the D-malate salt (3.7g, 4.93mmol) of (S) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml of ethyl acetate, stirring for 1 hour at room temperature, carrying out suction filtration to remove salt, and concentrating an ethyl acetate mother liquor under reduced pressure to obtain (R) -2- ((4- ((7-chloroquinolin-4-yl) (methyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (1.1g), wherein the purity is more than 99%, the chiral purity is more than 99.93%, and the yield is 44%.

Example 49: preparation of L-malate salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

The racemate 2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (5g, 13.33mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), L-malic acid (7.1g, 5.33mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of L-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete, dissolving most of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol L-malic acid salt in ethanol/ethyl acetate mixed solution, and L-malic acid salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol was precipitated, and filtered under suction to give a crude L-malic acid salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (5.0 g).

Example 50: recrystallization purification of L-malate salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Weighing a crude L-malate salt (5.0g) of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol, suspending the crude L-malate salt in 10mL of ethanol/ethyl acetate (v/v ═ 1:2), heating and refluxing to completely dissolve, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, further cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain an L-malate salt (4.6g) of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol with the optical purity of 87.3 percent ee.. Then, the salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol obtained above was subjected to one-time recrystallization according to the above-mentioned recrystallization method to obtain an L-malate salt of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (4.0g) having an optical purity of ee. (98.9%).

Example 51: preparation of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Sodium methoxide (1.1g,20.59mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, and L-malate salt (4.0g, 5.15mmol) of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml of ethyl acetate, stirring at room temperature for 1 hour, carrying out suction filtration to remove salt, and concentrating an ethyl acetate mother liquor under reduced pressure to obtain (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (1.3g), wherein the purity is more than 99%, the chiral purity is more than 99.90%, and the yield is 52%.

Example 52: preparation of D-malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

The racemate 2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (5g, 13.33mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 30mL), D-malic acid (7.1g, 5.33mol) was dissolved in ethanol/ethyl acetate (v/v ═ 1:2, 15mL), and the ethanol/ethyl acetate solution of D-malic acid was slowly added dropwise to the ethanol/ethyl acetate solution of the above racemate at 0 ℃. Heating to 5 deg.C, reacting for 30min until the reaction is complete, dissolving most of (R) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol salt of D-malic acid in ethanol/ethyl acetate mixed solution, and D-malic acid salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol was precipitated, and filtered under suction to give a crude D-malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (5.2 g).

Example 53: recrystallization purification of D-malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Weighing a crude D-malate salt (5.2g) of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol, suspending the crude D-malate salt in 10mL of ethanol/ethyl acetate (v/v ═ 1:2), heating and refluxing to be completely dissolved, slowly cooling to room temperature, keeping the temperature and stirring for 1-2 h, further cooling to 5-10 ℃, stirring for 30min, and performing suction filtration to obtain a salt (4.7g) of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol with the optical purity of 87.3 percent ee.. Then, the salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol obtained above was subjected to primary recrystallization according to the above-mentioned recrystallization method to obtain a D-malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (4.2g) having an optical purity of ee. (98.9%).

Example 54: preparation of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol

Sodium methoxide (1.2g,21.62mol) was added in portions to 10ml of anhydrous methanol, and after stirring at 0 ℃ for about 10min, the system became clear, the D malate salt of (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (4.2g, 5.40mmol) was slowly added, stirred at room temperature for 30min, and concentrated under reduced pressure to remove methanol. Adding 20ml of ethyl acetate, stirring at room temperature for 1 hour, carrying out suction filtration to remove salt, and concentrating an ethyl acetate mother liquor under reduced pressure to obtain (S) -2- ((4- ((7-chloroquinolin-4-yl) (cyclopropyl) amino) pentyl) (ethyl) amino) ethyl-1-ol (1.4g), wherein the purity is more than 99%, the chiral purity is more than 99.91%, and the yield is 56%.

Example 55: test experiment for cytotoxicity and anti-influenza virus drug effect of test compound

Test compound cytotoxicity assay: the cytotoxic effect of the compound on Vero cells is detected by adopting an MTT method, which is also called as an MTT colorimetric method and is a method for detecting the survival and growth of the cells. MTT (yellow thiazole blue) can penetrate through cell membranes to enter cells, amber dehydrogenase in mitochondria of living cells can enable exogenous MTT to be reduced into water-insoluble needle-shaped Formazan crystals and deposited in the cells, the crystals can be dissolved by dimethyl sulfoxide (DMSO), an enzyme linked immunosorbent detector is used for detecting the light absorption value at the wavelength of 490nm/570nm, and the quantity of the living cells can be indirectly reflected.

1) Inoculating Vero cells into a 96-well plate, and adding 100 mu L of cell suspension into each well; the number of cells per well was about 100000 cells in 5% CO₂Incubating and culturing at 37 ℃ for 24 h;

2) setting drug concentration gradient, setting 3 multiple wells for each concentration gradient, diluting the drug into corresponding culture medium to desired final concentration, sucking out original culture medium in 96-well plate, adding prepared culture medium containing drug with desired final concentration 100 μ L in 5% CO₂Incubating at 37 ℃; simultaneously setting a blank group (only containing 100 mu L of culture medium, no cells and the same subsequent treatment as other wells) and a control group (containing cells and culture medium);

3) 10 μ L of MTT solution (5mg/ml) was added to each well at 44 hours of drug treatment and incubation was continued for 4h (48 hours total drug treated cells);

4) the well was aspirated to remove the medium (if the cells were suspended, the medium was aspirated after centrifugation at 2500rpm for 5 min). Add 150. mu.L of DMSO to each well and shake until the crystals are fully dissolved. Detecting the light absorption value of each hole at OD 490nm on an enzyme-labeling instrument;

5) calculating an inhibition rate: the inhibition ratio is 1- (the OD value of the drug addition agent-the OD value of the blank group)/(the OD value of the control group-the OD value of the blank group) — (the OD value of the control group-the OD value of the drug addition agent)/(the OD value of the control group-the OD value of the blank group);

6) IC calculation Using Graphpad software₅₀A value;

the anti-influenza activity of the compound to be tested is determined by a plaque inhibition method, and the specific operation steps are as follows:

1) vero cells were seeded in 96-well plates (5X 10)⁵Cells/well) at 5% CO₂Incubating overnight at 37 ℃;

2) the influenza virus strain A/PR/8/34 is diluted to 100PFU/mL virus solution by adopting a serum-free culture medium, the virus solution (0.5 mL/hole) is added into a 96-hole plate, and Vero cells are infected for 1 h;

3) after completion of inoculation and adsorption, the virus culture was discarded and the cells were washed 1 time with PBS (pH 7.2);

3) to a 96-well plate, 80% agar, 1% bovine serum albumin and 3 μ g/mL acetyl trypsin, and an EMEM medium were added to dilute the test compound and the positive control plavir (8 concentrations were set, 3 duplicate wells per concentration) to the desired concentrations, and additionally, a virus control well (virus-infected cells, no test compound) and a normal cell well (normal cells, no test compound) were set. After the mixture is completely condensed, turning over the mixture up and down, and culturing for 72 h;

4) removing agarose after the culture is finished, and fixing the cell layer for 3min by using 70% ethanol at room temperature; removing ethanol, adding 1mL of crystal violet coloring agent, and dyeing at room temperature for 5 min; removing the crystal violet stain and washing the cells twice with PBS; the number of plaques was then calculated. The plaque inhibition was expressed as a percentage compared to the control against the virus;

5) EC was calculated using GraphPad software₅₀A value;

6) the final Selection Index (SI), SI ═ IC, of the compound was calculated₅₀/EC₅₀；

Specific experimental results are shown in table 1, the compound in the example has low cytotoxicity, and the inhibition activity and the selection index of (R) configuration chloroquine, hydroxychloroquine and analogues thereof and (S) configuration chloroquine, hydroxychloroquine and analogues thereof on influenza virus infection are superior to those of raceme chloroquine, hydroxychloroquine and analogues thereof, and the compound has better selection index;

TABLE 1 cytotoxicity, anti-influenza Activity and selection index

Example 56: activity assay against novel coronavirus pneumonia (COVID-19)

The specific operation steps of the detection of the anti-novel coronavirus pneumonia activity of the compound to be detected are as follows:

1) vero cells were seeded in 96-well plates (5X 10)⁴Cells/well) at 5% CO₂Incubating overnight at 37 ℃;

2) adding a novel coronavirus (MOI ═ 0.05) into a 96-well plate, infecting Vero cells for 2h, and discarding virus liquid;

3) adding a test compound containing a culture medium diluted to a required concentration and a positive control method Pilarvir (8 concentrations are set, each concentration is 3 multiple wells), and additionally arranging a virus control well (cells infected with virus and not containing the test compound) and a normal cell well (normal cells and not containing the test compound); after continuing culturing for 48h, collecting cell supernatant, adopting a lysis buffer solution for lysis, and using the lysis buffer solution for subsequent qRT-PCR determination of RNA copy number;

4) collecting 100. mu.L cell supernatant, and extracting virus RNA according to MiniBEST virus RNA/DNA extraction kit instructions; RNA was eluted using RNase-free water. Reverse transcription was performed using PrimeScript RT Reagent kit containing gDNA Eraser, and qRT-PCR was performed using StepOne Plus Real-time PCR system and TB Green Premix Ex Taq II.

5) Removing genome DNA in 3 mu L of total RNA by using gDNA scanner, and then synthesizing first strand cDNA in 20 mu L of reaction solution, wherein 2 mu L of cDNA is used as a template for quantitative PCR; using cDNA as a template, and adopting primers used for PCR amplification of a receptor domain of the Spike gene: RBD-F:5 '-GCTCCATGGCCTAATATTACAAACTTGTGCC 3' and RBD-R: 5'-TGCTCTAGACTCAAGTGTCTGTGGATCAC-3', then cloning into pMT/BiP/V5-His vector, and using the vector as plasmid standard after sequencing confirmation;

6) the primer of the quantitative PCR is RBD-qF1: 5'-CAATGGTTTAACAGGCACAGG-3', RBD-qR1:5 ' -CTCAAGTGTCTGTGGATCACG-3; reaction conditions are as follows: pre-denaturation at 95 ℃ for 5min, denaturation at 95 ℃ for 15s, annealing at 54 ℃ for 15s, extension at 72 ℃ for 30s, and reaction cycle number of 40 Cycles;

7) calculating copy number according to standard curve, and calculating EC by GraphPad software₅₀A value;

8) the final Selection Index (SI) of the compound, SI ═ EC, was calculated₅₀/IC₅₀；

Specific experimental results are shown in table 2, wherein the inhibitory activity and selection index of chloroquine and hydroxychloroquine of (R) configuration and chloroquine and hydroxychloroquine and analogs of (S) configuration on influenza virus infection are superior to those of racemic chloroquine and hydroxychloroquine and analogs thereof, and the selection index is better;

TABLE 2 cytotoxic, anti-novel coronavirus pneumonia Activity and selection index

Sequence listing

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