CN101717398B - Sophocarpine-series derivative as well as preparation method and purposes thereof - Google Patents

Abstract

The invention relates to a sophocarpine-series derivative as well as and a preparation method and purposes thereof. The sophocarpine-series derivative is characterized in that the general formula of the sophocarpine-series derivative is (I), wherein the R is CH3MgBr, EtMgBr, n-PrMgBr, i- PrMgBr, n-BuMgBr, i- BuMgBr, n-C5H11MgBr, i- C5H11MgBr, n-C6H13MgBr, n-C7H15MgBr, n-C12H25MgBr, and the like. The invention also discloses the preparation method and the purposes of the sophocarpine-series derivative. The preparation method relates to the diversity synthesis and the activity research of a structure based on sophocarpine, the structures of sophocarpine molecules are reformed and derived in an organically chemical synthesizing method, and then an antiviral medicament with high efficacy and low toxicity is obtained. The sophocarpine-series derivative has strong and obvious biologic activities, and has a very important function on relieving the pain of extensive patients and improving the living quality of the patients as a novel antiviral medicament with high efficacy and low toxicity.

Description

Sophocarpine series derivates and preparation method thereof and purposes

Technical field

The present invention relates to a kind of Chinese medical extract sophocarpine series derivates, the invention still further relates to the preparation method and its usage of this sophocarpine series derivates.

Background technology

Viral myocarditis is the primary myocardial inflammation caused by close Cardiovirus, often involves pericardium, causes perimyocarditis.So-called idiopathic myocarditis is because virus infection causes most probably.Myocarditic viral species can be caused a lot of, and wherein modal is COxsackie (Coxsackie) virus, echo, enteric cytopathogenic human orphan virus (i.e. the lonely virus of people's intestines), rubella virus, influenza virus, mumps virus etc.

The myocarditis of the mankind infects the most common with Coxsackie B virus group.Generally speaking, close Cardiovirus directly can destroy myocardial cell, but also indirectly destroys myocardial cell by the immune response of T cell mediation.Because the glycoprotein molecule structure of this type of viral capsid is similar to the glycoprotein of myocardial cell membrane, therefore after infection, antibody that body produces (antibody of activating complement and the antibody of neutralization virus) both for virus, also for myocardial cell.Therefore, after cytotoxic T cell is by sensitization, can destroy by the myocardial cell of virus infection.Viral myocarditis is one of common heart trouble, is mainly in person between twenty and fifty, mostly causes for CBV.Pump failure, the high Atrioventricular Conduction group of can happening suddenly is stagnant, Ah's spira's syndrome or form irregular pulse fast and die suddenly.As recurrent exerbation, develop into dilated cardiomyopathy, 75% is dead in onset 5 years, and consequence is very serious.

Sophocarpine is the effective constituent in kuh-seng, and in Herba Sophorae alopecuroidis, content is higher, and extracts easier.It has multiple biology, physiologically active.Sophocarpine central nervous system, cardiovascular systems, antiviral, relieving inflammation and relaxing pain, immunity and antitumor etc. in all there is clear and definite pharmacological activity and application prospect, have also been obtained extensive application clinically, Chen Shuxia etc. report that sophocarpidine has the effect of unique anti-Coxsackie B_3 virus (CVB), wherein the strongest with the effect of sophocarpine anti-Ke Saji B group 3 type (CVB3), and Ke Saji B3 virus is the myocarditic main pathogen of human viral.Sophocarpine has obvious anti-inflammatory action to the animal acute exudative inflammation that the multiple proinflammatory agent such as Oleum Tiglii, carrageenin brings out.Its anti-inflammatory action and hypophysis-adrenal system have nothing to do, and may be by direct acting mode.Therefore the means such as organic synthesis technology and biology synthetic drugs are utilized, based on similarity principle, study by structure activity relationship, filter out novel structure, biological action is unique, the active compound that the pharmacy of medicine, medicine generation, drug effect, security reach optimum matching is significant and good feasibility.Ren Ji hospital of Shanghai Second Emdical University entrusts national new medicament screen research centre in the screening to anti-SARS virus activity, find that sophocarpine is when the 100 above concentration of μ g/mL, there is obvious anti-severe acute respiratory syndrome (severe acuterespiratory syndrome, SARS) effect of virus, (its EC50 is about 127 μ g/mL).Recently sophocarpine injection (being used for the treatment of CBV myocarditis) has obtained State Food and Drug Administration enters clinical study certification about Chinese medicine first kind new drug.But but slowly great progress is not obtained for the research of sophocarpine structural derivative.

At present all over the world, virus disease, as: the global prevalence of SARS, bird flu, mad cow disease, acquired immune deficiency syndrome (AIDS), viral hepatitis etc., makes the research of antiviral become the focus of people's extensive concern.Think at present: in crowd, the morbidity of viral myocarditis is 2.3% ~ 5.0%, in newborn infant, children's sudden death case, 20% caused by viral myocarditis, and the same period accounts for 8.69% ~ 20.80% in cardiovascular disorder inpatient.Virus infection is except causing acute injury, and some patients prognosis is not good, can cause dilated cardiomyopathy, even in heart failure; Severe myocarditis patient also likely occurs extremely broken.Antiviral why slower development, major cause is that the cycle of virus is in close relations with the metabolic process of host normal cell, suppresses the damage that viral breeding certainly will cause non-infected cells.Constantly find along with to virus-specific drug target target, and the appearance of a large amount of specificity antivirus medicine, people recognize gradually can selective therapy virus infection, has both suppressed viral proliferation and don't has affected the growth of host cell.The main policies of present antiviral research comprises: the combination of blocking virus and target cell, suppress reversed transcriptive enzyme, suppress viral protein translation, arrestin matter is modified or sprouting of virion discharges and cut off viral genome etc. in addition, also have by carrying out structure of modification to known curative effect compound, by bioinformatics software design guide molecule, the general sour structure of modification of adopted few core and chemosynthesis, immunomodulator development and screen natural anti-virus lead compound etc. from plant and microorganism.But, think effective to treatment of viral myocarditis at present and clinical medicine can be applied to little.

Summary of the invention

Technical problem to be solved by this invention is for the deficiencies in the prior art, provide a kind of newly, antiviral activity is high, toxicity is low sophocarpine series derivates.

Another technical problem to be solved by this invention there is provided a kind of preparation method of sophocarpine series derivates as above.

Another technical problem to be solved by this invention there is provided a kind of purposes of sophocarpine series derivates as above.

Technical problem to be solved by this invention is realized by following technical scheme.The present invention is a kind of sophocarpine series derivates, is characterized in, it has following general formula (I),

Wherein, R is CH ₃mgBr, EtMgBr, n-PrMgBr, i-PrMgBr, n-BuMgBr, i-BuMgBr, n-C ₅h ₁₁mgBr, i-C ₅h ₁₁mgBr, n-C ₆h ₁₃mgBr, n-C ₇h ₁₅mgBr, n-C ₁₂h ₂₅mgBr, BnMgBr,

Technical problem to be solved by this invention can also be realized further by following technical scheme.Present invention also offers a kind of preparation method of the sophocarpine series derivates as described in above technical scheme, be characterized in, its reaction equation is as follows,

Reaction conditions is:

General formula (II) is sophocarpine, and R is CH ₃mgBr, EtMgBr, n-PrMgBr, i-PrMgBr, n-BuMgBr, i-BuMgBr, n-C ₅h ₁₁mgBr, i-C ₅h ₁₁mgBr, n-C ₆h ₁₃mgBr, n-C ₇h ₁₅mgBr, n-C ₁₂h ₂₅mgBr, BnMgBr,

Below the correlation test that does of contriver and result thereof.

1, testing compound:

1), compound 1:

Chemical name: 16-methyl-15,16-seco-matridin-15-oic acid methyl ester

Molecular formula: C ₁₇h ₃₀n ₂o ₂

Structural formula:

2), compound 2:

Chemical name: (S)-13-methyl-matridin-15-one;

Molecular formula: C ₁₆h ₂₆n ₂o;

Structural formula:

2. Drug toxicity trails:

Testing compound is added in the HeLa cell containing cell maintenance medium by following concentration respectively: 8,16,32,125,250,500,2000,4000,8000 μ g/ml, each concentration establishes 4 repeating holes, in 37 DEG C, 5%CO ₂cultivate in incubator, observation of cell toxicity change day by day.Calculate cell survival rate.

Cell survival rate=(experimental port OD value/control wells OD value) × 100%

3. In vitro antibacterial test (CPE suppresses method and mtt assay):

Testing compound RPMI1640 substratum is added after 1: 2 to 1: 128 doubling dilutions 40 orifice plates (final concentration 1.875 ~ 120 μ g/mL), each extent of dilution adds 4 holes, with 100TCID ₅₀cVB3m suspension is placed in CO together ₂1h in incubator, it is 3 × 10 that every hole adds concentration again ⁵the HeLa cell suspension of/mL, puts CO ₂incubator is cultivated.Establish virus, sophocarpine and cell controls group simultaneously.Cultivate and observe CPE on the 4th day.Survey the OD value at 570nm place, each hole with mtt assay when virus control, " +++ ~ ++++", and by following formulae discovery to the inhibiting rate of CVB3.

Viral suppression=(drug treating group OD value-virus control group OD value)/(cell controls group OD value-virus control group OD value) × 100%

Therapeutic index (TI)=TC50/IC50, calculates TC50 and IC50 by the Probit Return Law of SPSS11.5.The results are shown in Table 1 and table 2.

Table 1 medicine is to caused CPE restraining effect and viral suppression

Group	Drug dose (μ g/mL)	Cytopathy (CPE)	Viral suppression (%)
				Sophocarpine	1.875	++	10.4
	3.75	++	21.7
					7.5	+	42.8
	15	+	52.4
					30	+	64.2
	60	++	70.1
					120	++	75.9
16-methyl-15，16-seco-matridin-15-oic acid methyl ester	1.875	+++	30.5
					3.75	+++	44.6
	7.5	++	51.8
					15	++	58.3
	30	+	65.9
					60	+	74.2
	120	－	85.9
				(S)-13-methyl-matridin-15-one	1.875	+++	33.8
	3.75	+++	47.1
					7.5	++	52.3
	15	++	58.8
					30	+	66.2
	60	+	78.9
					120	－	88.2
Cell controls group		－
				Virus control group		++++

Table 2 medicine is to the toxicity of cell

Cell survival rate	Sophocarpine	16-methyl-15，16-seco-matridin-15-oic acid methyl ester	(S)-13-methyl-matridin-15-one
				Drug dose (μ g/mL)
8	100	100	100
				16	100	100	100
32	100	100	100
				125	78.6	91.3	96.5
250	65.2	82.5	82.6
				500	41.6	76.5	72.3
2000	10.5	55.8	63.2
				4000	2.8	39.2	55.9
8000	1.2	22.4	36.9
				IC50	400.3287±62.0648	2151.582±499.299	3821.019±1148.26

Result shows, and 16-methyl-15,16-seco-matridin-15-oic acid methyl ester and (S)-13-methyl-matridin-15-one have obvious provide protection to the cell that external CVB3 infects.Its median effective dose is respectively 7.2636 ± 2.33415,6.2147 ± 2.04095 μ g/ml, and median toxic dose is then 5.37,9.55 times of sophocarpine respectively.Prompting 16-methyl-15,16-seco-matridin-15-oic acidmethyl ester has definite antivirus action with (S)-13-methyl-matridin-15-one and toxicity is lower, and security is higher than sophocarpine.

Pharmacodynamics in vitro screening and toxicity test: Coxsackie B virus 3 (CVB3) is one of main inducing of viral myocarditis.Therefore the target that screens using CVB3 as Anti-viral activity in vitro of this research, set up the viral myocarditis cell model that CVB3 infects HeLa cell and myocardial cell, select ribavirin and sophocarpine as positive control medicine, and measure the toxicity of each derivative compound at cell levels.By cytopathic effect inhibition (CPE) experiment, viral proliferation Inhibition test, preliminary screening is carried out to the antiviral activity of compound.

The present invention is that the structure diversity based on sophocarpine synthesizes and activity research, by using organic chemical synthesis means to derive the transformation that sophocarpine molecule carries out structure, obtains antiviral that is efficient, low toxicity.Sophocarpine series derivates of the present invention has stronger obvious biologic activities, and it is as a kind of antiviral that is novel, high-efficiency low-toxicity, and to the misery alleviating extensive patients, the quality of life improving patient has very important effect.

Embodiment

Below further describe concrete technical scheme of the present invention, so that those skilled in the art understands the present invention further, and do not form the restriction to its right.

Embodiment 1.A kind of sophocarpine series derivates, it has following general formula (I),

Wherein, R is CH ₃mgBr, EtMgBr, n-PrMgBr, i-PrMgBr, n-BuMgBr, i-BuMgBr, n-C ₅h ₁₁mgBr, i-C ₅h ₁₁mgBr, n-C ₆h ₁₃mgBr, n-C ₇h ₁₅mgBr, n-C ₁₂h ₂₅mgBr, BnMgBr,

Its preparation method reaction equation is as follows,

Reaction conditions is:

General formula (II) is sophocarpine, and R is CH ₃mgBr, EtMgBr, n-PrMgBr, i-PrMgBr, n-BuMgBr, i-BuMgBr, n-C ₅h ₁₁mgBr, i-C ₅h ₁₁mgBr, n-C ₆h ₁₃mgBr, n-C ₇h ₁₅mgBr, n-C ₁₂h ₂₅mgBr, BnMgBr,

Claims (2)

1. a sophocarpine derivative, is characterized in that, it has following structural formula:

2. the purposes of sophocarpine derivative as claimed in claim 1 in the anti-CVB3 virus drugs of preparation.