CN1315176A - Application of bilirubin bitaurine sodium and its bilirubin derivative in preparing anti-HIV medicine - Google Patents

Abstract

An application of bilirubin bitaurine sodium and its bilirubin derivative in preparing anti-HIV medicine is disclosed. Experiments prove that they have the effect on suppressing HIV without any toxicity. The bilirubin bitaurine sodium has the highest effect.

Description

Sodium bilirubin ditaurate and its bilirubin derivatives as preparation of anti-HIV drugs

The invention belongs to the application of bilirubin derivatives in the field of medicine, and in particular relates to the use of bilirubin derivatives, especially sodium bilirubin ditaurate (Ditaurobilirubin, DTB) as an anti-HIV drug.

AIDS (AIDS), Acquired Immunodeficiency Syndrome, is a very serious infectious disease that threatens human health in the world. By the end of 1999, there were a total of 49.9 million HIV-infected and AIDS patients worldwide, of which 33.6 million were still alive and 16.3 million had died of AIDS. HIV began to be introduced into my country through importing foreign blood products (1982-1984). By the first half of 1999, the Chinese government reported that 13,619 people were infected. Chinese experts estimated that the number of infected people exceeded 400,000. The World Health Organization estimated that my country's 1998 There are 600,000 infected people, which will reach 800,000 in 1999 and 1.2 million in 2000.

The first case was discovered in the United States in 1981. In 1983, Professor Montagnier of the Pasteur Institute in France first isolated the virus from the cells cultured in the patient's lymph nodes, which was later named HIV (Humanimmunodeficiency virus, HIV), thus proving that AIDS It is a viral infectious disease characterized by acquired immunodeficiency. Because HIV loves _CD4 lymphocytes, on the one hand, it continuously reproduces and releases in the cells, and the released virus invades new lymphocytes; on the other hand, _CD4 lymphocytes invaded by the virus can interact with other _CD4 cells. Fusion forms a giant nuclear syncytium, and the syncytia itself is unstable and easily leads to cell death. The virus reproduces, releases, syncytia forms, and dies. Repeatedly, it causes the body's deep cellular immune deficiency, and finally destroys the immune function of the human body and leads to the patient's death. In addition to invading _CD4 cells, HIV can also invade macrophages and B lymphocytes. In particular, it forms chronic infection in macrophages, and HIV can exist in the body for a long time. The development of the disease can be divided into four stages: acute infection, asymptomatic infection, pre-AIDS and AIDS. Once the infected person develops into AIDS, various pathogens can cause opportunistic infections and different tumors, leading The patient eventually died of exhaustion.

In 1985, it was discovered that azidothymidine (AZT, 3'-Azidothymidine) has the activity of inhibiting reverse transcriptase in vitro, and clinical research was carried out in 1986. In 1987, AZT was the first to be approved by the US Food and Drug Administration (FDA). , used for the treatment of AIDS, the application shows that it can inhibit the replication of HIV and prolong the survival time of patients for half a year to one year. It also has serious side effects of bone marrow suppression, and it is easy to produce drug resistance. It cannot eliminate hidden dangers and is expensive. General patients are often unaffordable, limiting its long-term widespread use. Subsequently, the US FDA has successively approved AZT similar drugs dideoxycytidine (ddC, 2', 3'-Dideoxycytidine), dideoxyinosine (ddl, 2', 3'-Dideoxyinosine) and dideoxythiocytidine (3 -TC, 2'3'-Dideoxy-3'-thiacytidine), several drugs are used in rotation clinically, but the problem of toxicity and drug resistance cannot be completely solved, and the life is prolonged for two or three years. Since 1995, the AIDS research group led by Dr. He Dayi in the United States has used several drugs in combination therapy on the basis of studying protease inhibitors, called Highly Active Antiretroviral Therapy (HAART). The reverse transcriptase and protease parts in the life cycle of the virus can be used in combination to prolong the life span of 2/3 patients to 4-5 years, and it is expected to be longer. Although this method can obviously prolong the patient's life, it still has three disadvantages: ① Drug-resistant HIV strains will still appear, which has toxic side effects; 3. It is inconvenient to take, and the amount is large (dozens of capsules per day), and it needs to be taken in sequence under the guidance of a doctor at any time. In addition, the research time of this method is still short, and its long-term effect needs to be further evaluated.

Therefore, it is still very important to search for new anti-HIV drugs from various ways. The Japanese had reported in 1991 that biliverdin had anti-HIV effects, see the article (H.Mari et.al., Jpn.J.Cancer Res.82, 755 -757,1991); the article of University of California, San Francisco in "Journal of Biological Chemistry" 1996, volume 320, pages 681-686 (F.McPheeet, al., Biochem.J.320,681-686,1996) believes that bilirubin is Promising HIV protease inhibitors. The Institute of Photosensitive Chemistry, Chinese Academy of Sciences has conducted research on bilirubin for many years and synthesized various derivatives. The Institute of Virology of the Chinese Academy of Preventive Medicine began experimental research on anti-HIV drugs in 1988.

The object of the present invention is to show that bilirubin ditaurine sodium (DTB) and other bilirubin derivatives inhibit HIV, and provide a use for preparing a medicine for treating AIDS.

The purpose of the present invention is achieved like this:

The structure of the bilirubin derivative used in the technical scheme of the present invention is as shown in formula (I):

Wherein R ₁ , R ₂ , R ₃ , R ₆ , R ₇ , R ₈ are alkyl, vinyl or alkylcarbonyl; R ₄ , R ₅ are fatty acid groups, sodium salts, ammonium salts of fatty acids or pharmacologically acceptable Acceptable salt: or a substituent containing sulfonic acid, sodium salt of sulfonic acid, ammonium salt or pharmaceutically acceptable salt; _R8 can also be a substituent containing sulfonic acid, sodium salt of sulfonic acid, ammonium salt or pharmaceutically acceptable salt acceptable salt substituents;

When the substituent is the following group, it is the following compound:

R ₁ R ₂ R ₃ R ₄ R ₅ R ₆ R ₇ R ₈ Sodium Bilirubin Ditaurate (DTB) M V M PS PS M M M Bilirubin ⅩⅢα(MBR) M E M P P M E M Bilirubin ⅩⅢα Sulfonic Acid Derivatives M E M S S M E M Bilirubin Side Group Derivatives M V M P P M M ES

Among them, M=CH ₃ , V=CH:CH ₂ , PS=CH ₂ CH ₂ CONHCH ₂ CH ₂ SO ₃ H, its sodium salt, ammonium salt or pharmaceutically acceptable salt; E=CH ₂ CH ₃ , P=CH ₂ CH ₂ COOH, its sodium salt, ammonium salt, pharmaceutically acceptable salt or ditaurine derivative; S=(CH ₂ )nSO ₃ H, its sodium salt, ammonium salt or A pharmaceutically acceptable salt wherein n=1-3; ES= _CH2CH ( _CH3 ) _CH2CH2SO3H , its sodium salt, ammonium _salt or a pharmaceutically acceptable salt _.

The structure of another class of bilirubin derivatives used in the technical scheme of the present invention is shown in formula (II):

Wherein R ₁ , R ₂ , R ₃ are alkyl, vinyl or alkylcarbonyl; R ₄ is fatty acid, sodium salt of fatty acid, ammonium salt of fatty acid or pharmaceutically acceptable salt, or sulfonic acid, sulfonic acid Substituents of sodium salts, ammonium salts or pharmaceutically acceptable salts.

When the substituent is the following group, it is the following compound:

R ₁ R ₂ R ₃ R ₄

Yellow Bilirubic Acid (XBR) M E M P

Taurine Derivatives of Yellow Bilirubic Acid M E M PS

Where M=CH ₃ , E=CH ₂ CH ₃ , P=CH ₂ CH ₂ COOH, its sodium salt, ammonium salt or pharmaceutically acceptable salt; PS=CH ₂ CH ₂ CONHCH ₂ CH ₂ SO ₃ H , its sodium salt, ammonium salt or pharmaceutically acceptable salt. The structure of used sodium bilirubin ditaurate (DTB) in the technical scheme of the present invention is as shown in formula (Ⅲ):

where M=CH ₃ ; V=CH:CH ₂ ; PS=CH ₂ CH ₂ CONHCH ₂ CH ₂ SO ₃ Na

Bilirubin ditaurine sodium salt (DTB) physical and chemical properties: molecular weight 798,9 (Na-free), yellow solid, soluble in water, dimethyl sulfoxide, slightly soluble in alcohol, insoluble in non-polar organic solvent. It is stable in solid state or in alkaline solution, but easily oxidized in acidic solution.

The literature has reported its maximum absorption and molar absorptivity ε in protein buffer, see Table 1. See "Clinical Chemistry", Volume 31, 1985, pages 1677-1682 (B.T.Doumas et.a1., Clin.Chem.31, 1677-1682, 1985).

Table 1 Maximum absorption and molar absorptivity of DTB

εx10 ^-3 Lmol ^-1 cm ^-1 Tris(pH8.5) Phosphate buffer (pH7.4) BSA (bovine serum albumin) HSA (human serum albumin) Human serum (human serum) Bovine serum (bovine serum) H ₂ O (450nm) (460nm)56.755.357.257.5-- (422nm)50.4----47.9-- (460m)43.255.958.945.749.5

Preparation of sodium bilirubin ditaurate (DTB):

1. Bilirubin (Bilirubin IXα)

Bilirubin is purchased from the market. Because the purity is too low, it must be purified and recrystallized. The purification method refers to "Journal of Biochemistry" 1972, 129 pages 797-800 (AFMcDonagh and F.Assisi, Biochem.J., 129797-800, 1972) and makes improvements. The specific method is as follows: add 500 mg of commercially available bilirubin into 500 mL of destabilized chloroform, heat to reflux under nitrogen flow for half an hour, and then cool to room temperature. The solution was filtered and the chloroform solution was washed with 0.1M NaHCO ₃ (3×100 mL). Then filter through double filter paper to remove water in chloroform. The chloroform solution was concentrated by rotary evaporation under vacuum at 30 °C. Then add methanol to the concentrated chloroform solution until it becomes turbid, put it in the refrigerator until crystals appear, and filter. Wash with methanol containing a little chloroform, then dry under vacuum. According to different sources of raw materials, the recovery rate is 50-90%, and the purity is more than 98% (calculated by the molar extinction coefficient of 65000), and the whole operation should be carried out in the dark. All solvents were pre-gassed with argon for half an hour.

2. Synthesis of Sodium Bilirubin Ditaurate (DTB)

The starting materials we used were bilirubin and taurine sodium salt. Combine bilirubin and taurine according to the principle of peptide synthesis. Its chemical reaction scheme is as follows, and synthetic method is as follows: bilirubin (1.2g) adds triethylamine 0.6mL in ethyl acetate, ethyl chloroformate 0.45mL, stirs 2 hours, then adds taurine sodium salt ( NH ₂ CH ₂ CH ₂ SO ₃ Na) methanol solution (0.51g taurine sodium salt in 20mL methanol), stirred well, overnight, centrifuged, and dried to obtain solid 1.5g (yield 87%), in methanol and Recrystallize in ethyl acetate and dry in vacuum to obtain the product DTB. The yield of recrystallization is 98%, and the purity is greater than 95%. Confirmation of the structure of DTB

We confirmed the chemical structure of DTB by conventional physical methods.

①Infrared Spectrum The infrared spectrum of DTB is shown in Figure 1. Figure 1 is obtained by making potassium bromide pellets from DTB samples after twice recrystallization, purification and drying. The characteristic identification points are ①formation of amide: the first double frequency of bending vibration in the NH plane of amide 3300cm ^-1 ; the absorption band of amide Ⅰ is 1640cm ^-1 ; the absorption band of amide Ⅱ is 1560cm ^-1 ; ②the characteristic band of sulfonate is 1050cm ^-1 and 1200cm ^-1 . The remaining peaks closely resemble the IR spectrum of bilirubin itself.

②Visible absorption spectrum Figure 2 is the visible absorption spectrum of DTB in phosphate buffer containing human serum albumin at pH 7.4. Since the chromophore part of bilirubin does not change after being chemically bonded with taurine, the absorption spectrum is basically similar to that of bilirubin.

DTB合成路线图

DTB synthetic route map

③ NMR spectrum was measured on a 400MHz high-resolution NMR spectrometer using heavy water as a solvent. The main peaks and their attribution list 2 are as follows:

Table 2. 400MHz NMR spectrum data of DTB (internal standard TMS=0ppm) chemical shift Peak multiplicity and coupling constant Hydrogen number attribution 1.9151.9302.9273.1013.2673.3563.4833.5013.964 sst 6.8Hzt 6.8Hzt 6.8Hzt 6.8Hzt 6.8Hzt 6.8Hzm 662222224 Pyrrole CH ₃ Pyrrole CH ₃ -CH ₂ CH ₂ CONH--CH ₂ CH ₂ CONH--CH ₂ CH ₂ CONH--CH ₂ CH ₂ CONH--CH ₂ CH ₂ SO ₃ Na-CH ₂ CH ₂ SO ₃ Na _{-CH2CH2SO3Na _ _} Note: Due to the presence of a small amount of water in the sample and the purity limit of the deuterated reagent, the absorption intensity of the water peak (HDO at ~5ppm) in the NMR spectrum is particularly large, resulting in the low intensity of the olefin hydrogen itself due to coupling and other reasons, which cannot be very good The hydrogen on 10-CH ₂ is covered by the solvent peak, and 10-CH ₂ should be around 4.5ppm, which is not resolved in this spectrum.

④Mass Spectrometry Due to the large molecular weight of DTB, and there are many chemical bonds that are easy to break in the DTB molecule, the mass spectrum of DTB cannot be determined by electron bombardment (EI) method. Mass spectrometric analysis of DTB was performed using fast atom bombardment (FAB) negative ion mass spectrometry. The mass number of the negative ion peak obtained was 855. Its negative ion group may have the following structure:

Due to the rotation of 10-CH ₂ , the carbonyl groups on the skeleton of bilirubin can respectively form hydrogen bonds with matching water molecules to stabilize the negative ion groups.

From the above data analysis, it can be concluded that the chemical structure of the DTB prepared by us is correct. It is the chemical structure listed in the chemical reaction formula.

Purity analysis of DTB

① Absolute purity

The prepared DTB was crystallized twice, then vacuum-dried (1mmHg) by phosphorus pentoxide for 10 hours, and the absolute purity analysis of DTB was carried out.

② Relative content ratio

The prepared DTB was analyzed by high pressure liquid chromatography. Adopt C18 reverse-phase analytical column to detect at 450mm, do eluent with 92% 0.1mol n-octylamine methanol solution and 8% water, see " American Chemical Society Journal " 1982 volume 104 page 6865 (A.F.McDonagh, L.A.Palma , F.R.Trull and D.A.Lightner, J.Am.Chem.Soc., 104, 6865, 1982).

The structure of yellow bilerubic acid (XBR) used in the technical scheme of the present invention is as shown in formula (IV):

Wherein M=CH ₃ ; E=CH ₂ CH ₃ ; P=CH ₂ CH ₂ COOH The synthetic method of yellow bile rubic acid (XBR) is:

The oxime of ethyl acetoacetate and pentanedione are condensed into ethyl 4-acetyl-3,5-dimethyl-1hydropyrrole-2-carboxylate under the reduction of acetic acid-zinc powder, and further reduced, hydrolyzed and decarboxylated , oxidation, and bromination to obtain 5-bromomethylene-4-ethyl-3-methyl-2-oxo-2,5-dihydropyrrole, which is combined with 2,4-dimethyl-5-carboxy - 1-hydropyrrole-3-propionic acid is condensed under acid-catalyzed reflux in methanol to form yellow bile rubic acid methyl ester, and then hydrolyzed with NaOH aqueous solution to obtain yellow bile rubic acid (XBR). References "Journal of Heterocyclic Chemistry", Volume 21, 1984, pp. 139-144 (D.A. Lightner, J.S.Ma et.al., J. Heterocyclic Chem., 21 139-144, 1984.).

Structural parameters: IR(KBr)ν3360, 3200, 2500, 1705, 1670, 1630 cm ^-1 . ¹ H NMR (DMSO-d ⁶ ): δ1.10(t,3H),1.81(s,3H),2.07(s,3H),2.22(s,3H),2.25-2.80(m,6H),5.93( s, 1H), 9.67 (br s, 1H), 10.18 (broad s, 1H), 11.83 (brs, 1H). Uv-vis: λ _max (CHCl ₃ )=408nm, ε=28000, λ _max (CH ₃ OH)=416nm, ε=34000.

The structure of bilirubin XIIIα (MBR) is shown in formula (Ⅴ):

Where M=CH ₃ ; E=CH ₂ CH ₃ ; P=CH ₂ CH ₂ COOH, the synthesis method of bilirubin XⅢα is:

Yellow bile rubinate methyl ester is oxidatively condensed by 2,3-dichloro-5,6-dicyano-1,4-p-benzoquinone (DDQ) into biliverdin XIIIα dimethyl ester, and finally treated with sodium borohydride It can be reduced and hydrolyzed into medium bilirubin XIIIα with a purity of over 99%. References "Journal of Heterocyclic Chemistry", Vol. 24, pp. 1573-1579, 1987 (R.T. Trull, R.W. Franklin and D.A. Lightner, J. Heterocyclic Chem., 24 1573-1579 1987).

Structural parameters: IR (liquid film): ν3420, 3260, 2966, 1704, 1689, 1629 ^{cm -1} . ¹ HNMR (deuterated chloroform): δ1.12 (t, 6H, J=7Hz), 1.85 (s, 6H, 2, 18-position CH ₃ ), 1,85 (s, 6H, 7, 13-position CH ₃ ) ,2.48(q,4H,J=7Hz),2.78(m,8H,-CH ₂ CH ₂ COOH),4.06(s,2H,10-CH2-),6.04(s,2H,5,10= CH), 9.15 (s, 2H, pyrrole NH), 10.52 (s, 2H, lactam NH), 13.31 (s, 2H, COOH) ppm; MS: M ⁺ 587.7. UV-Vis (chloroform): λ _max =431 nm, ε = 60000. ¹³ CNMR (deuterated chloroform): 8.54(q), 9.62(q), 15.29(q), 17.64(t), 19.74(t), 23.96(t), 34.81(t), 98.19(d), 119.72( s), 122.43(s), 123.00(s), 123.39(s), 128.28(s), 130.81(s), 147.67(s), 172.42(s), 174.46(s).

The compound of the present invention can be used to prepare medicine for treating AIDS.

Configuration of bilirubin ditaurine sodium salt (DTB) anti-HIV drug reagent:

Dissolve bilirubin ditaurine sodium salt (DTB) directly in serum-free RPMI 1640 culture medium (0.5mg/mL) to prepare the desired concentration (see Table 3 and Table 4).

Configuration of yellow bilerubic acid (XBR) anti-HIV drug reagents:

Accurately weighed and prepared as a saturated solution of dimethyl sulfoxide (DMSO), the concentration was 12.3 mg/mL. During the experiment, it was diluted to the required concentration with serum-free RPMI 1640 medium (see Table 3 and Table 4).

The configuration of bilirubin ⅩⅢα (MBR) anti-HIV drug reagents:

Accurately weighed and prepared as a saturated solution of dimethyl sulfoxide (DMSO), the concentration was 11.6 mg/mL. During the experiment, it was diluted to the required concentration with serum-free RPMI 1640 medium (see Table 3 and Table 4).

The compounds of the present invention, especially DTB, can be prepared into capsules, tablets, injections and the like by conventional methods.

Effect of the present invention:

We screened a variety of bilirubin derivatives synthesized against HIV, and in vitro experiments proved that several bilirubin derivatives have the effect of inhibiting HIV, among which DTB has the best effect, and its half inhibitory concentration (IC ₅₀ ) was 3.75 μg/mL, the inhibition rate was 100% when the concentration was 40 μg/mL, and the inhibition rate was 90% when the concentration was 10 μg/mL. Cytotoxicity experiments showed that the cytotoxicity of DTB was zero. The acute toxicity test on mice shows that intragastric administration to mice is equivalent to more than 390 times the effective dose of antiviral in vitro, without any toxicity. DTB is a component of chicken and fish gallbladder, and is also used in traditional Chinese medicine. Using our synthetic method, it is easy to synthesize and produce in large quantities.

When these compounds, especially DTB, are prepared into capsules, tablets, and injections by traditional methods, they can achieve the same effect of inhibiting HIV.

These compounds, especially DTB, are used as anti-AIDS drugs in combination with AZT and the like, and can also achieve the same effect of inhibiting HIV.

Description of drawings:

Figure 1 - Infrared Spectrum of Synthetic DTB

Figure 2 - Visible absorption spectrum of synthesized DTB

The following examples further illustrate the pharmacological and toxicological effects of the pharmaceutical compounds of the present invention.

Embodiment 1 Anti-HIV virus experiment of the compound of the present invention

In this experiment, an in vitro method was used to observe the effect of drugs on inhibiting virus replication in cells after lymphocyte strains were infected with HIV-1. 1. Experimental Materials

The virus is HIV-1, which was donated by Professor Montagnier of the Pasteur Institute in France. The concentration of the virus used in the experiment was 1×10 ⁴ TCID ₅₀ /mL (TCID ₅₀ is 50% cytopathic dose); it was preserved in CEM cells HIV-1 virus was subcultured by the HIV laboratory of Institute of Virology, Chinese Academy of Preventive Medicine. Drugs use DTB, XBR and MBR. During the experiment, it was dissolved in serum-free RPMI1640 culture medium to prepare the required concentration (see Table 3 and Table 4). 2. Experimental method (1) Virus and cell culture:

Co-culture freshly cultured MT ₄ cells (5×10 ⁵ /mL) with HIV-1 virus liquid (10 ³ TCID ₅₀ /mL), act in a CO ₂ incubator at 37°C for 1-1.5 hours, and complete with RPMI 1640 After the culture solution (containing 10% calf serum and antibiotics) was used to wash away the free virus from uninfected cells, the cell concentration (5×10 ⁵ /mL) was corrected with the complete culture solution for further use. (2) Determination of HIV-1p24 antigen expression:

A micro-ELISA method is used to quantitatively measure the p24 antigen of HIV virus in the supernatant of cultured cells in vitro to indicate the amount of virus. Operate according to kit instructions.

Results According to the OD value, the Cut Off value and the standard curve, the amount of virus contained in each well (ie pg/mL) was calculated. (3) Anti-HIV-1 virus experiment, determine the half maximal inhibitory concentration (IC ₅₀ ) of the drug:

实验结果如下：The cell volume is 5×10 ⁵ /mL, add a certain amount of HIV-1 virus, act at 37°C for 60-90 minutes, centrifuge, discard the supernatant, add serum-free RPMI 1640 to suspend the cells, centrifuge as above and discard the supernatant RPMI 1640 culture medium with double serum was added to make 5×10 ⁵ /mL. Use a 96-well plate for the experiment, dilute the drug to different concentrations, set four reaction wells for each concentration, and set up virus control and cell control reaction wells, add 100 μL of drug and 100 μL of virus-infected cells to each well, and incubate at 37 °C After three days, the medium was changed, 100 μL of supernatant was removed from each well, and 100 μL of the same drug solution or culture medium as in the original well was added. After six days, the supernatant of each well was collected, and the virus contained in each well was determined according to the above-mentioned p24 antigen analysis method. The inhibition rate was calculated according to the following formula. According to the inhibition rate of each concentration group, calculate the half inhibitory concentration (IC ₅₀ ), and make statistics according to the formula.

The experimental results are as follows:

Table 3 Antiviral activity compound stock solution dilution Concentrationμg/mL OD value Inhibition rate XBR 1/2001/4001/800 43.2521.6310.81 0.0580.0880.112 80.56% 72.22% 47.22% MBR 1/2001/800 29.007.25 0.0680.100 78.89% 54.44% DTB 1/12.51/501/2001/800 40.0010.002.500.625 0.0150.0300.1320.147 100.0% 90.00% 43.33% 30.56% virus control group 2.5 _IC50 of DTB ≈3.75μg/mL (4) Cytotoxicity test:

MT ₄ cells were mixed with double-serum culture solution to make 5×10 ⁵ /mL, and planted in a 96-well plate, adding 100 μL to each well, and adding 100 μL of different concentrations of drug solution. In the control group, no drug was added, and no serum was added. 100 μL of RPMI 1640 nutrient solution, cultured at 37°C for three days, aspirated 100 μL of supernatant from each well, added 0.25% MTT (blue tetrazolium, prepared with pH7.2 phosphate buffer), 20 μL per well, and incubated at 37°C for 4 hours. The yellow MTT forms purple crystals under the action of the mitochondria of living cells, add 100 μL DMSO to each well to dissolve the crystals, then measure the optical density (OD value) with a wavelength of 570 nm on a microplate reader, and then calculate the cell killing rate according to the following formula : The experimental results are as follows:

Table 4 Cytotoxicity compound stock solution dilution Concentrationμg/mL OD value Killing rate XBR 1/2001/4001/800 43.2521.6310.81 0.2470.30650.3405 29.32% 12.30% 2.58% MBR 1/2001/4001.800 29.0014.507.25 0.260.3150.313 25.61% 9.87% 10.44 DTB 1/12.51/501/2001/4001/800 40.0010.002.501.250.625 0.3610.3700.3950.3720.492 0% 0% 0% 0% 0% cell control group 0.3495

Example 2 Toxicological experiment of the compound of the present invention

Acute toxicity test in mice:

Animals: Kunming outbred mice (provided by the Experimental Animal Breeding Field of the Institute of Zoology, Chinese Academy of Medical Sciences) were divided into males and females, weighing 18-20 grams, and the males and females were housed in separate cages. Before the experiment, the mice were fed and observed in the laboratory for 3-5 days free to eat and drink.

Drugs: DTB, dissolved in water.

Method: Mice were administered intragastrically, and the performance and death were observed after administration, and the mortality rate was recorded for three days. Results: The dosage of the mice was 25 mg/20 grams of body weight. After the administration, the mice had normal activities, no toxicity, and no death in three days. This dose was equivalent to more than 390 times the effective dose of antiviral in vitro.

Example 3 Sodium bilirubin ditaurate was made into capsules using known techniques in the art.

Example 4 Sodium bilirubin ditaurate was made into tablets using known techniques in the art.

Example 5 Sodium bilirubin ditaurate was made into an injection using known techniques in the art.

Claims (5)

1. Bilirubin ditaurate and bilirubin derivative thereof be as the purposes of preparation anti-hiv drug, it is characterized in that the application that chemical compound that following structural formula (I) or structure formula II are represented is used to prepare anti-hiv drug:

R wherein ₁, R ₂, R ₃, R ₆, R ₇, R ₈Be alkyl, vinyl or alkyl-carbonyl; R ₄, R ₅Be acceptable salt on sodium salt, ammonium salt or the materia medica of fatty acid-based, fatty acid; Or contain the substituent group of acceptable salt on sodium salt, ammonium salt or the materia medica of sulfonic acid, sulfonic acid;

Or

R wherein ₁, R ₂, R ₃Be alkyl, vinyl or alkyl-carbonyl; R ₄Acceptable salt or contain the substituent group of acceptable salt on sodium salt, ammonium salt or the materia medica of sulfonic acid, sulfonic acid on the sodium salt of fatty acid, the ammonium salt of fatty acid or the materia medica.

2. a kind of Bilirubin ditaurate as claimed in claim 1 and bilirubin derivative thereof is characterized in that the R of the chemical compound that described structure formula I is represented as the purposes of preparation anti-hiv drug ₈The application that is used to prepare anti-hiv drug during for the substituent group of acceptable salt on the sodium salt, ammonium salt or the materia medica that contain sulfonic acid, sulfonic acid.

3. a kind of Bilirubin ditaurate as claimed in claim 1 and bilirubin derivative thereof is characterized in that chemical compound that described structure formula I or structure formula II are represented is used to prepare anti-hiv drug when substituent group is following group application as the purposes of preparation anti-hiv drug:

R ₁R ₂R ₃R ₄R ₅R ₆R ₇R ₈The pendant derivatized thing M of DTB M V M PS PS M M V mesobilirubin X III α M E M P P M E M mesobilirubin X III α sulfonic acid M E M S S M E M bilirubin V M P P M M E S

Wherein, M=CH ₃, V=CH:CH ₂, PS=CH ₂CH ₂CONHCH ₂CH ₂SO ₃Acceptable salt on H, its sodium salt, ammonium salt or the materia medica; E=CH ₂CH ₃, P=CH ₂CH ₂Acceptable salt or two taurine derivatives on COOH, its sodium salt, ammonium salt, materia medica; S=(CH ₂) nSO ₃Acceptable salt, wherein n=1-3 on H, its sodium salt, ammonium salt or the materia medica; ES=CH ₂CH (CH ₃) CH ₂CH ₂SO ₃Acceptable salt on H, its sodium salt, ammonium salt or the materia medica;

Or

When substituent group is following group, be following chemical compound:

R ₁???R ₂???R ₃???R ₄

Xanthobilirubic acid M E M P

The taurine derivatives M E M PS of Xanthobilirubic acid

M=CH wherein ₃, E=CH ₂CH ₃, P=CH ₂CH ₂Acceptable salt on the sodium salt of COOH, ammonium salt or the materia medica;

PS=CH ₂CH ₂CONHCH ₂CH ₂SO ₃Acceptable salt on H, its sodium salt, ammonium salt or the materia medica.

4. as the described a kind of Bilirubin ditaurate of claim 1-3 and bilirubin derivative thereof the purposes as the preparation anti-hiv drug, it is characterized in that being prepared into capsule, tablet, injection is the application of anti-AIDS drug.

5. as the described a kind of Bilirubin ditaurate of claim 1-4 and bilirubin derivative thereof purposes, it is characterized in that being used as the application of anti-AIDS drug with the zidovudine compatibility as the preparation anti-hiv drug.

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