CN1035767C

CN1035767C - Arteannuin derivant and its composition and application

Info

Publication number: CN1035767C
Application number: CN93108651A
Authority: CN
Inventors: 罗宣德; 李泽林; 马林
Original assignee: Individual
Current assignee: Individual
Priority date: 1992-11-10
Filing date: 1993-07-19
Publication date: 1997-09-03
Anticipated expiration: 2013-07-19
Also published as: CN1102413A

Abstract

The present invention relates to an arteannuin medicine for resisting AIDS viruses. Dihydro-arteannuin methylamino formate is an effective medicine for resisting AIDS viruses, which has the advantages of low toxicity, large therapeutic index, high safety and reliability and low cost. The arteannuin medicine is a medicine firstly invented in China, which has a novel structure. The medicine is a novel derivative of arteannuin, and has prominent progress in treating AIDS.

Description

New artemisinin derivative and uses thereof

The present invention relates to the isocyclic compound field, the derivative dihydro-abrotine methylamino-manthanoate that particularly artemisine of sesquiterpene lactones is new.

Acquired immune deficiency syndrome (AIDS) (AIDS) is that acquired immunodeficiency syndrome (Acquired immune deficiencysyndrome) was found first case in the U.S. in 1981, nineteen eighty-three, France's bus moral professor Montagnier of institute at first isolated virus from patient blood, later called after HIV, thereby the proof acquired immune deficiency syndrome (AIDS) be with the day after tomorrow acquired immunodeficiency be the viral infectious of feature, because virus of AIDS (HIV) parent has a liking for the T lymphocyte, it is constantly bred, discharges in this cell on the one hand, and the virus that discharges is invaded new T lymphocyte again; The T4 lymphocyte of being invaded by virus can merge with other T4 lymphocytes and form huge nuclear synplasm on the other hand, and the instability of synplasm own easily causes death.Viral proliferation, release, synplasm form, death, so therefore cause the cellular immunization of the body degree of depth damaged repeatedly, finally destroy immune function of human body and cause death.Virus of AIDS especially forms chronic infection except that to also invading scavenger cell, bone-marrow-derived lymphocyte etc. the T lymphocyte in scavenger cell, but the virus long-term existence.This sick development can divide the carrier, and in sick three stages of ARC phase and AIDS, in case develop into the AIDS stadium, morbidity is rapid, and three annual survival rates are less than 10%.Have 2,000 ten thousand people to infect virus of AIDS at present in the world approximately, AIDS stadium patient reaches 6,000,000, has half people dead approximately.China was no exception, and acquired immune deficiency syndrome (AIDS) has been imported China in 1984, present HIV the infected's 890 examples of having found of China, and Chinese carrier's 740 examples wherein, AIDS phase patient 5 examples, the HIV patient is still constantly increasing.

Anti-hiv drug, that first is reported is Suramine (Suramin), found that AZT (Azidothymidine) etc. had the activity of external anti AIDS virus in 1985, carried out clinical study in 1986,1987AZT is used for the treatment of the medicine of acquired immune deficiency syndrome (AIDS) as first by drugs approved by FDA.So far in the world wide screening hundreds of kind new compound and prescription thereof arranged, and comprise tens kinds of natural drugs and Chinese medicine.Up to the present, only AZT, DDI (didanosine), DDC (zalcitabine) of the medicine that is used for the treatment of acquired immune deficiency syndrome (AIDS) by drugs approved by FDA; Trichosanthin (GLQ in natural drug and the Chinese medicine ₂₃) carried out clinical observation by the FDA approval, but these several medicines all exist dissimilar toxicity, use 4-6 after week as AZT, and bone marrow depression appears in patient, develops into serious anaemia then; Can produce resistance after separately using 6 months, and AZT can not there be restraining effect to the virus that infects in the scavenger cell, thereby can not removes a hidden danger, cost an arm and a leg simultaneously.DDC and DDI exist peripheroneural toxicity, promptly occur after 6 weeks of taking medicine, and use heavy dose, even drug withdrawal still had sequela in 1 year.Trichosanthin has been found neural poison in clinic trial, momentary dementia appears in severe patient, even stupor, and experimental study shows the scavenger cell of Trichosanthin processing infected by HIV, produce and release solubility toxicant, people's brain cell is had serious destruction.

Up to the present worldwide being removed seldom in hundreds of kind medicine and prescription, natural drug, single medicinal material and the herbal mixture by having of experimental study and evaluation, number has separated effective constituent, as: Japan's report glycyrrhizin, lentinan etc., the Philippine Violet Herb effective constituent of California, USA university report etc., and number average experimentizes with crude extract mostly, mostly be nucleosides in the compound, adenosine or peptide derivative.And to 1992 in world's acquired immune deficiency syndrome (AIDS) conference that Amsterdam, the Netherlands (Amsterdam) is held, do not see that yet drug research has breakthrough.

The object of the invention is: provide a kind of chemical effective constituent of from Chinese medicine, extracting Dihydroqinghaosu-methylcarbamate they are low, the inexpensive anti-AIDS class medicines of toxicity.Overcome the shortcoming of AZT, DDI, DDC medicine.

Another object of the present invention is the composition with this compound anti-AIDS-virus.A further object of the invention is with this compound or contains the application of the pharmacological agent AIDS disease of this compound.

The objective of the invention is to reach like this: adopted China to have the Chinese medicine sweet wormwood of affluent resources, it is the Artemisinin (artemisinin) that extracts in the plant Artemisia annua (Artemisia annua L.), and then the synthesizing series derivative particularly Dihydroqinghaosu-methylcarbamate make a breakthrough using of anti AIDS virus, thereby the object of the invention has just reached fully.

Below the present invention is described in detail.

Medicine Dihydroqinghaosu-methylcarbamate structure of the present invention is as follows:

Dihydroqinghaosu-methylcarbamate

This derivative forms by the Dihydroartemisinin derivation.Synthesis preparation method is as follows:

1. Dihydroartemisinin: Artemisinin is dissolved in methyl alcohol, adds sodium borohydride, and after react, with the acid neutralization, reaction solution adding saturated brine is separated out crystallization, the crystallization of separation dry raw, recrystallization again, yield about 80%.

2. Dihydroqinghaosu-methylcarbamate

The solution of Dihydroartemisinin 284mg (1mmol) in methylene dichloride 6ml adds methyl isocyanate 63mg (1.1mmol) stirring and refluxing and gets white crystals mp175-177 ℃ of (sherwood oil: ethyl acetate)

Ultimate analysis: meet C ₁₇H ₂₇NO ₆1/2H ₂O (350.42)

C H N

Theoretical value (%) 58.27 8.05 4.00

Actual value (%) 58.48 0.09 4.08

In the present composition, formula (I) compound accounts for the 0.1-5% of said composition weight, preferred 1-2%.

The present invention estimates external anti-AIDS toxic action with this artemisine compounds.One, the experiment of anti HIV-1 virus

The method of the evaluation inverase of generally acknowledging is in the world adopted in experiment: 1. experiment material

Virus is HIV-1, and this virus is that the Montagnier of Pasteur Institut professor gives, and used virus titer is 1 * 10 in the experiment ⁴TCID ₅₀/ ml;

Cell with cem cell preserving HIV-1 virus, by the cultivation of going down to posterity of this laboratory.

Medicine Dihydroartemisinin, methylamino-manthanoate Artesunate.Positive control drug AZT, liquor strength are 1mg/ml.2. experimental technique

A. with the MT4 cell or the U937 cell (5 * 10 of fresh culture ⁵Ml) with viral liquid (10 ⁴TCID ₅₀/ ml) the common cultivation, 37 ℃ of effects are 1-1.5 hour in the CO2 incubator, with RPMI 1640 train fully basic (containing 10% calf serum and antibiotic) flush away not with cell bonded virus, standby to train basic corrected concentrations fully.

Test in 96 orifice plates and carry out, add the MT4 cell suspension of the above-mentioned infective virus of 0.1ml in every hole, add the soup of 0.1ml different concns then.Other establishes positive drug AZT control group, also establishes the virus control (the MT4 cell that virus infection is only arranged) of no soup simultaneously, and the soup of each concentration is established two holes.Test group and control group are cultivated in 37 ℃ of following CO2 incubators, and the liquid of changing dressings after three days carries out following experimental observation after six days;

B cell growing state is observed; Respectively organize amount of viable cell with tire Finland staining (Trypan blue dye) observation;

The mensuration of c virus P24 antigen presentation: check the virus antigen expression with immunoenzyme, promptly respectively organizing cell with the slide glass of band circular hole is applied in the hole respectively, each drug level is coated with two holes, cold acetone is fixed, and drip anti-HIV positive serum, in 37 ℃ of following CO2 incubators, cultivate after 30 minutes, it is inferior to give a baby a bath on the third day after its birth with PBS, dripping enzyme mark SPA and above-mentioned similarity condition again cultivated 30 minutes, it is inferior to give a baby a bath on the third day after its birth with PBS, places substrate solution dyeing 2-3 minute then, and distilled water washes down slide, examine under a microscope, normal cell is that the colourless cell that has virus is red-brown.3. the result judges

Virus control group smear is seen many tangible red-brown cells.

AZT1 * 10 ^-1* 10 ^-21 * 10 ^-3Each group loses pink cell fully and represents that virus is suppressed, and shows that method is reliable, and used mark is as follows:

＂-＂ represents full Kong Weijian positive cell;

＂ ± ＂ represents that only there is 1-2 probable positive cell in full hole;

There is 2-3 positive cell in the full hole of "+" expression;

Experiment repeats two to three times.

Each medicine stoste is that the viruliferous cell suspension of 0.1ml adding 0.1ml is got in the 1mg/ml experiment, and pretend with concentration as follows: the stoste group is that 0.1mg/0.2ml is 0.5mg/ml, 1 * 10 ^-1Group is 0.05mg/ml, 1 * 10 ^-2Group is 0.005mg/ml1 * 10 ^-3Also 0.5ug/ml is by that analogy for 0.0005mg/ml for group.

Table 1 medicine concentration 1 * 10 ^-11 * 10 ^-21 * 10 ^-31 * 10 ^-4Two blue or green hydrogen artemisin methylamino-manthanoate---±

AZT - - - /

So the Dihydroqinghaosu-methylcarbamate minimal effective concentration is 1 * 10 ^-4Be that activity is 0.05ug/ml.At each concentration group cell well-grown of Artesunate, compare no significant difference with the cell control group.

Activeconstituents of the present invention can be mixed with the dosage form of any routine, comprises solid form and liquid form, and solid form can be a tablet for example, capsule, suppository, minigel, pulvis, particle looses, transdermal ointment, and navel applies cream etc., liquid preparation such as oral liquid, close liquid, oil for injection injection, the powder pin, liquid drugs injection, intramuscular injection or infusion etc.

Accompanying drawing is the mass spectrum of The compounds of this invention.

Claims

1, formula (I) compound that has following structure:

2, the application of compound as claimed in claim 1 in the medicine of preparation treatment AIDS.