CN109336800B - DHNB (dehydroepiandrosterone) condensed phenylthiosemicarbazide compound as well as preparation method and application thereof - Google Patents
DHNB (dehydroepiandrosterone) condensed phenylthiosemicarbazide compound as well as preparation method and application thereof Download PDFInfo
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- C07C337/00—Derivatives of thiocarbonic acids containing functional groups covered by groups C07C333/00 or C07C335/00 in which at least one nitrogen atom of these functional groups is further bound to another nitrogen atom not being part of a nitro or nitroso group
- C07C337/06—Compounds containing any of the groups, e.g. thiosemicarbazides
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Abstract
The invention discloses a chemical synthesis drug DHNB (dehydroepiandrosterone) condensed phenylthiosemicarbazide, which has the following structure:
the preparation method of the chemical synthetic medicine DHNB (dehydroepiandrosterone) thiosemicarbazone takes 3, 4-dihydroxy-5-nitrobenzaldehyde (DHNB) shown in a structural formula (I) as a raw material, and the DHNB thiosemicarbazone shown in a structural formula (III) is synthesized by condensation reaction of the DHNB thiosemicarbazone and 4-phenylthiosemicarbazone shown in a structural formula (II) in a solvent methanol through aldehyde amine; the invention can effectively treat gout and hyperuricemia by inhibiting Xanthine Oxidase (XOD) and reducing in vivo uric acid synthesis.
Description
Technical Field
The invention relates to the field of new drug development and application, in particular to a chemical synthesis drug DHNB (dehydroepiandrosterone) thiosemicarbazone, a preparation method thereof and application thereof in preventing and treating gout and hyperuricemia.
Background
Gout is a crystal-related arthropathy caused by deposition of monosodium urate (MSU) and is directly related to Hyperuricemia (HUA) caused by purine metabolic disorder or reduced uric acid excretion. The prevalence rate of the HUA in China is on the rise year by year, the onset age of the HUA is more and more low, the HUA becomes the second most metabolic disease next to diabetes, and huge economic and mental burdens are brought to the society and families. Epidemiological research in the last decade shows that the prevalence rate of the HUA in different areas of China is greatly different and is 5.46-19.30%, wherein 9.2-26.2% of men and 0.7-10.5% of women are in the number of the men, and the total number of the men is higher than that of the women. Gout is a group of clinical syndromes of tissue damage caused by long-term purine metabolic disorder (or) reduction of uric acid excretion and increase of blood uric acid, and hyperuricemia is the most important biochemical basis of gout. Uric acid is produced in the liver by dietary intake and purine compounds decomposed in the body, and about 2/3 of uric acid is excreted through the kidney, and the rest is excreted from the digestive tract. Gout comprises acute gouty arthritis and chronic tophus diseases, and the rise of blood uric acid can cause gout and is also related to the occurrence and development of systemic diseases such as kidney, endocrine metabolism, heart, cerebral vessels and the like.
At present, the conventional medicaments for treating gout mainly comprise colchicine, non-steroidal anti-inflammatory drugs, allopurinol, febuxostat, benzbromarone and the like. The medicines can inhibit the formation of uric acid or (and) promote the excretion of uric acid to achieve the aim of relieving and treating gout and hyperuricemia. The synthesis of uric acid in the body is associated with purine metabolism, and Xanthine Oxidase (XOD), a key enzyme in uric acid production in the body, catalyzes the conversion of hypoxanthine and xanthine into uric acid. Hyperuricemia in the body can be caused by hyperuricemia, and further gout can be caused. Allopurinol and febuxostat are inhibitors of XOD, and the two drugs can reduce synthesis of uric acid in vivo by inhibiting XOD, so that gout is effectively treated. However, they all have certain side effects in clinical use, for example, allopurinol can cause skin anaphylaxis and damage to liver and kidney functions, and severe patients can generate hypersensitivity syndromes such as lethal exfoliative dermatitis and the like; febuxostat can cause liver function impairment, nausea, rash, and the like. Adverse reactions of taking benzbromarone include gastrointestinal discomfort, diarrhea, rash and liver function damage; adverse reactions of colchicine include nausea, vomiting, diarrhea, abdominal pain, liver dysfunction, renal function injury and the like. Although more medicines for preventing and treating gout and hyperuricemia exist at present, most medicines have large side effects and influence the quality of life. Therefore, the development of new drugs for treating hyperuricemia and gout is still a hot spot of research in the medical field.
Research shows that 3, 4-dihydroxy-5-nitrobenzaldehyde (DHNB) belongs to a derivative of protocatechualdehyde, is a strong XOD inhibitor, and can be used as a potential drug for treating hyperuricemia and gout. DHNB can reduce serum uric acid levels in hyperuricemic mice, and a large dose of DHNB has no side effect on the mice (500 mg/kg). However, DHNB has the defect of time dependence on inhibition of XOD activity, has extremely short action time, and has weaker inhibition capability on XOD than allopurinol.
Disclosure of Invention
The invention aims to overcome the defects of the prior art, adapt to practical development and provide a chemical synthesis medicament DHNB (dehydroepiandrosterone) phenylthiosemicarbazone, a preparation method thereof and application of the DHNB phenylthiosemicarbazone in preventing and treating gout and hyperuricemia so as to solve the problems in the background technology.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a chemical synthetic drug DHNB (dehydroepiandrosterone) thiosemicarbazone has the following structure:
the preparation method of the chemical synthesis medicine DHNB (dehydroepiandrosterone) condensed phenylthiosemicarbazide takes 3, 4-dihydroxy-5-nitrobenzaldehyde (DHNB) shown in a structural formula (I) as a raw material, and the DHNB condensed phenylthiosemicarbazide and 4-phenylthiosemicarbazide shown in a structural formula (II) undergo a condensation reaction in a solvent methanol through aldehyde amine to synthesize the DHNB condensed phenylthiosemicarbazide shown in a structural formula (III), wherein the molar ratio of the 3, 4-dihydroxy-5-nitrobenzaldehyde (DHNB) shown in the structural formula (I) to the 4-phenylthiosemicarbazide shown in the structural formula (II) is 1.0-1.05; the reaction formula of the above process is shown below.
The application of the chemical synthesis medicine DHNB (dehydroepiandrosterone) thiosemicarbazone in preventing and treating gout and hyperuricemia is that the DHNB thiosemicarbazone is used as a pharmaceutical active ingredient to be prepared into tablets, capsules, granules, injection, liposome or sustained-release controlled-release preparations.
The DHNB phenylthiosemicarbazone can also be used in combination with allopurinol, febuxostat or colchicine for preventing and treating gout and hyperuricemia.
The DHNB phenylthiosemicarbazone can also be chelated with metal ions to form a corresponding metal complex for preventing and treating gout and hyperuricemia.
The application of the chemical synthetic medicine DHNB (dehydroepiandrosterone) thiosemicarbazone in preventing and treating gout and hyperuricemia is used as an additive to be applied to health-care food or feed.
Compared with the prior art, the invention has the following advantages and positive effects:
the invention provides a chemical synthetic drug DHNB (dehydroepiandrosterone) thiosemicarbazone, which is subjected to in vivo and in vitro pharmacodynamic experiments, wherein the in vivo experiments comprise the determination of the influence of the thiosemicarbazone on serum XOD, liver XOD and serum uric acid of a hyperuricemia model mouse; in vitro experiments, a multifunctional microplate reader is used for measuring the inhibition effect of the enzyme label on XOD, and the inhibition mechanism is discussed; and carrying out acute toxicity test to obtain the high-efficiency and low-toxicity DHNB phenylthiosemicarbazone.
The research on the in vitro inhibition of XOD enzyme activity of the DHNB phenylthiosemicarbazone compound shows that: in a xanthine oxidase system, fixing the concentration of xanthine, changing the concentration of the xanthine oxidase, adding DHNB (dehydroepiandrosterone) thiosemicarbazone with different concentrations, and respectively measuring the inhibition effect of the DHNB thiosemicarbazone with different concentrations on XOD (X-ray diffraction order); with the increase of the concentration of the DHNB (dehydroepiandrosterone) thiosemicarbazone, the inhibition effect of the DHNB thiosemicarbazone compound on the XOD (X-ray dehydrogenase) enzyme activity is enhanced, namely the inhibition rate is in positive correlation with the concentration; and DHNB phenylthiosemicarbazone was more pronounced than DHNB and the positive control allopurinol.
The research on the influence of the DHNB (dehydroepiandrosterone) thiosemicarbazone compound on hyperuricemic model mice in vivo indicates that: the DHNB (dehydroepiandrosterone) phenylthiosemicarbazone can obviously reduce the activity of the serum XOD (x-ray oxygen deficiency) and the serum uric acid level of a hyperuricemia model mouse and can obviously reduce the activity of the liver XOD of the hyperuricemia model mouse; the inhibition of XOD activity and the capability of reducing serum uric acid are stronger than that of a positive control allopurinol.
Drawings
FIG. 1 is a chemical synthesis scheme of DHNB phenylthiosemicarbazone according to the present invention;
FIG. 2 shows the inhibition of different concentrations of XOD by DHNB thiosemicarbazone;
FIG. 3 is a Lineweaver-Burk plot of DHNB phenylthiosemicarbazone inhibition of XOD catalysis;
FIGS. 4 to 5 show the results of the inhibition of serum XOD activity of mice with hyperuricemia by DHNB-p-phenylamino thiourea;
FIGS. 6 to 7 show the effect of DHNB (dehydroepiandrosterone) on blood uric acid of a hyperuricemic mouse model;
FIG. 8 shows the results of the inhibition of liver XOD activity by DHNB-dephenylthiosemicarbazone in hyperuricemic mouse model.
Detailed Description
The following detailed description of specific embodiments of the invention refers to the accompanying drawings.
Preparation, separation and purification of DHNB (dehydroepiandrosterone) thiosemicarbazone compound
A chemical synthetic drug DHNB (dehydroepiandrosterone) thiosemicarbazone has the following structure:
the preparation method of the chemical synthesis medicine DHNB (dehydroepiandrosterone) thiosemicarbazone comprises the following steps:
taking 0.005mol of 4-phenylthiosemicarbazide in a 500mL three-neck flask, adding 80mL of methanol, heating and refluxing until the 4-phenylthiosemicarbazide is completely dissolved; dissolving 0.005mol of DHNB in 100mL of hot methanol; after 4-phenylthiosemicarbazide is completely dissolved, slowly dropping methanol liquid with DHNB dissolved into a three-neck flask by using a constant-pressure dropping funnel, dropping within 20min, dropping 3 drops of glacial acetic acid into the three-neck flask as a catalyst after dropping, continuously heating and refluxing for 2h, tracking and monitoring by TLC (thin layer chromatography), and completing the reaction until the raw material point disappears;
the specific reaction formula is shown as follows:
separation and purification of the chemical synthesis medicament DHNB phenylthiosemicarbazone: and (3) distilling and concentrating the DHNB phenylthiosemicarbazone methanol reaction liquid (reddish brown liquid) under reduced pressure, cooling to separate out crystals, filtering, washing with glacial acetone, and drying to obtain brick-red flocculent crystals.
In vitro inhibition of xanthine oxidase by DHNB phenylthiosemicarbazone
2.1 Experimental materials DHNB phenylthiosemicarbazone, phosphate Buffered Saline (PBS), xanthine Oxidase (XOD), allopurinol, and the like.
2.2 Experimental methods
2.2.1 enzyme Activity detection and sample inhibition XOD determination
The reaction temperature was 25 ℃ and the total reaction volume was 0.3mL (including 1mM EDTA in 0.05M pH =7.5 PBS buffer solution, 0.15mM xanthine substrate solution, 0.5 IU. ML) -1 XOD enzyme solution) was zeroed with buffer and measured with a multifunctional microplate reader containing enzyme kinetics software. Setting a normal group (without adding a sample) and a sample group in an experiment, sequentially adding 100uL of xanthine substrate solution and the sample solution into a 96-well plate, replacing the others with PBS (phosphate buffer solution), keeping the volume (300 uL) of the whole system consistent, finally adding 3uL of XOD solution to start reaction, measuring the absorbance value at 290nm, recording once at an interval of 10s, measuring the absorbance change value within 2min, taking time as an independent variable and the absorbance value as a dependent variable, obtaining an absorbance-time straight line, and calculating the slope Rate (dA/min) of the straight line. Each sample requires 3 parallel operations, and the inhibition rate of the sample is calculated by taking an average value. Inhibition (%) = [ (K1-K2)/K1]X100%. Wherein K1 represents the slope of the line of the normal group, and K2 represents the line of the sample groupThe slope. And calculating a regression equation by using the sample concentration as an independent variable X and the inhibition rate as a dependent variable Y and adopting an SPSS19.0 statistical software package. The concentration of the sample at which the inhibition rate was 50%, that is, the half-inhibitory concentration IC was calculated from the regression equation 50 。
2.2.2 Mechanism of inhibition of XOD by DHNB-Phenylthiosemicarbazone
Referring to 2.2.1, in the activity measuring system, the concentration of the substrate xanthine is fixed, DHNB phenylthiosemicarbazone is added at different concentrations, the mass concentration of XOD is changed, and the influence of inhibitors at different concentrations on the xanthine catalytic oxidation capability of XOD is measured. Plotting the enzyme concentration with the speed of the enzymatic reaction, and if a series of straight lines passing through the origin are obtained, the reversible inhibition is obtained; if a set of parallel lines is obtained, irreversible inhibition is indicated.
2.2.3 Inhibition type of DHNB (dehydroepiandrosterone) phenylthiosemicarbazone on XOD (x-ray diffraction)
Fixing the mass concentration of XOD, changing the concentration of substrate xanthine, and determining the influence of inhibitors with different concentrations on enzyme activity. By the Lineweaver-Burk equation: 1/V = Km/Vmax 1/[ S ] +1/Vmax plot can derive the inhibition type.
2.3 results of the experiment
2.3.1 Inhibition of xanthine oxidase by DHNB-condensed phenylthiosemicarbazide
Inhibition rate of DHNB, DHNB phenylthiosemicarbazone and positive control allopurinol on XOD and IC 50 As shown in table 1.
Table 1 results of inhibition of xanthine oxidase by different samples (n =3,
)
table 1 shows that the inhibition rates of different samples on XOD are different, the inhibition rates are in positive correlation with the concentrations, and the IC of DHNB on XOD 50 32.12. Mu. Mol/L of DHNB phenylthiosemicarbazone IC 50 Is 0.05 mu mol/L, has more remarkable inhibition effect than DHNB, and inhibits XOD activitySpecific allopurinol (IC) 50 =6.75 μmol/L) is stronger.
2.3.2 Inhibition mechanism of DHNB-phenylthiosemicarbazone on XOD
The results of inhibition of xanthine by DHNB-dephenylthiosemicarbazide at different concentrations are shown in FIG. 2, where four straight lines of inhibition of xanthine by xanthine oxidase at concentrations of DHNB-dephenylthiosemicarbazide of 0. Mu.g/mL, 0.01. Mu.g/mL, 0.02. Mu.g/mL, and 0.03. Mu.g/mL, respectively, are not parallel but intersect at one point, indicating that DHNB-dephenylthiosemicarbazide is a reversible inhibitor of xanthine oxidase.
2.3.3 Inhibition type of DHNB (dehydroepiandrosterone) phenylthiosemicarbazone on XOD (x-ray diffraction)
In the xanthine oxidase system, under the same XOD concentration condition, the volume of the substrate xanthine in the total 300 μ L (50 μ L (3.8 μ g/mL), 60 μ L (4.564 μ g/mL), 70 μ L (5.32 μ g/mL), 80 μ L (μ g/mL), 100 μ L (7.6 μ g/mL)) is changed, namely the mass concentration of the substrate xanthine in the system is changed, and the inhibition effect of the inhibitor DHNB thiosemicarbazone on the xanthine oxidase at 290nm is measured by a microplate reader at different concentrations (0 μ g/mL, 0.01 μ g/mL, 0.02 μ g/mL, 0.03 μ g/mL). Lineweaver-Burk curves were plotted as the inverse of the slope of the line versus the corresponding substrate concentration. As shown in FIG. 3, the Lineweaver-Burk curves for the inhibition of xanthine oxidase by DHNB phenylthiosemicarbazide have a common intersection in the first quadrant, and the intersection does not fall on a coordinate axis, indicating that the inhibition mechanism of DHNB phenylthiosemicarbazide against xanthine oxidase is neither competitive nor anti-competitive, and thus it can be preliminarily determined that the inhibition mechanism of DHNB phenylthiosemicarbazide against xanthine oxidase may be mixed.
Effect of DHNB Phenylthiosemicarbazone on hyperuricemic mice
3.1 the experimental animals are selected from 60 healthy male Kunming mice with the body weight of 18-22g, and provided by the scientific and technological center of the experimental animals of Jiangxi traditional Chinese medicine university (the production permit number of the experimental animals: SCXK (gan) 2018-003). Raising for one week before experiment to adapt to environment. Feeding conditions are as follows: the room temperature is 25 +/-2 ℃, and the relative humidity is 60-70%.
3.2 liquid medicine preparation Each group of the drug for intragastric administration and the animal molding substance was mixed with 0.9% CMC-Na solution to prepare drug-containing suspension.
3.3 Experimental methods
3.3.1 animal Molding
Potassium oxonate and uric acid ip mice are used for increasing serum uric acid level, so that a mouse hyperuricemia model is created. After the male Kunming mice are adaptively fed for one week, the mice are randomly divided into 6 groups, namely a normal saline group, a hyperuricemia model group, a DHNB phenylthiosemicarbazone low-dose group, a DHNB phenylthiosemicarbazone medium-dose group, a DHNB phenylthiosemicarbazone high-dose group and an allopurinol group, and each group comprises 10 mice. Except the normal saline group, other groups were modeled with Potassium Oxonate and uric acid ip mice for 2 days in succession to increase serum uric acid levels, resulting in a mouse hyperuricemia model.
3.3.2 Experimental procedures
The normal saline group and the hyperuricemia model group are both 20 mL/kg per day -1 Gavage (ig) dose contained 0.9% CMC-Na saline, 6 days consecutively, allopurinol group (10 mg. Kg) -1 ) Ig administration 1 time per day for 6d, DHNB phenylthiosemicarbazone low dose group at 5 mg-kg -1 Dose, DHNB Protophenylthiosemicarbazone Medium dose group at 10mg kg -1 Dose, DHNB Phenylthiosemicarbazone high dose group at 20mg kg -1 The dosage is 1 time per ig administration for 6 days, from the 5 th day of administration, except for the physiological saline group, the other groups need to be ip oteracil potassium and uric acid 1h before ig administration, and the dosages of the oteracil potassium and the uric acid are respectively 0.3 g.kg -1 And 0.25 g.kg -1 The molding was performed 1 time per day for 2 consecutive days. Collecting blood from femoral artery of mouse after 1h of ig administration on day 6, placing the blood sample in 1.5mL centrifuge tube, coagulating in refrigerator at 4 deg.C for 2h, at 3000 r.min -1 Centrifuging at low temperature for 5min. Serum uric acid levels and serum XOD viability were determined for each blood sample serum. After blood is taken, the liver of the dissected mouse is wrapped by the tin foil paper, and then is quickly frozen by liquid nitrogen and placed in a refrigerator at the temperature of 20 ℃ below zero to be tested for the activity of XOD.
3.1.3 Biochemical index detection
Mice were assayed for serum uric acid levels, blood and liver XOD activity, respectively. The blood uric acid of each group of mice is detected by a phosphotungstic acid method, and the XOD activity is detected by an enzyme colorimetric method. The specific operation is carried out according to the kit instruction.
3.2 results of the experiment
3.2.1 Inhibition effect of DHNB (dehydroepiandrosterone) phenylthiosemicarbazone on serum XOD (x-ray activating cell) activity of hyperuricemia model mouse
The activity of mouse serum XOD in a normal saline group, a high uric acid model group, a DHNB (dehydroepiandrosterone) thiosemicarbazone low-dose group, a DHNB (dehydroepiandrosterone) thiosemicarbazone medium-dose group, a DHNB thiosemicarbazone high-dose group and an allopurinol group is detected by adopting a phosphotungstic acid method, as shown in figure 4, the activity of the model group XOD and a normal saline control group show extremely obvious difference, and the success of experimental model building is shown. After modeling, the medium dosage of DHNB (dehydroepiandrosterone) thiosemicarbazone, the high dosage of DHNB thiosemicarbazone and the positive medicine allopurinol can inhibit the activity of mouse serum XOD (x-ray deficiency and blood pressure); the allopurinol group shows significant difference compared with the model group; compared with a model group, the DHNB phenylthiosemicarbazone shows very significant difference in the dosage and the high dosage. The results show that the DHNB (dehydroepiandrosterone) thiosemicarbazone has a good inhibition effect on the activity of mouse serum XOD (x-ray activating cell).
In addition, as shown in fig. 5, the doses of DHNB phenyl thiosemicarbazone and the high doses showed significant differences compared to the positive allopurinol, indicating that DHNB phenyl thiosemicarbazone has better effect of inhibiting XOD activity than allopurinol. The inhibition effect of different doses of DHNB (dehydroepiandrosterone) phenylthiosemicarbazone on the XOD (serum oxygen demand) activity of mice is different, and the inhibition effect of the low dose of DHNB phenylthiosemicarbazone on the XOD activity is poor; compared with allopurinol, the inhibition effect of the moderate and high dose of DHNB (dehydroepiandrosterone) thiosemicarbazone on the activity of mouse serum XOD (X-ray activating oxygen) is very significant and different; at the same time, there was no significant difference between the dose and the high dose in DHNB thiosemicarbazone. The results show that the DHNB phenylthiosemicarbazone has obvious inhibition effect on mouse serum XOD and dose correlation, and the total dose level in the DHNB phenylthiosemicarbazone is the best.
3.2.2 Effect of DHNB (dehydroepiandrosterone) phenylthiosemicarbazone on blood uric acid of hyperuricemia model mouse
After the mice are modeled by high uric acid, the serum uric acid levels of the mice in a normal saline group, a high uric acid model group, a DHNB (dehydroepiandrosterone) thiosemicarbazone low-dose group, a DHNB thiosemicarbazone medium-dose group, a DHNB thiosemicarbazone high-dose group and an allopurinol group are detected. As shown in FIG. 6, the serum content of the model group mice is very significantly different from that of the normal saline control group, which indicates that the experimental modeling is successful. The DHNB (dehydroepiandrosterone) thiosemicarbazone and the positive allopurinol can reduce the serum uric acid level of mice, and have significant difference compared with a model group. Mouse serum uric acid levels were lower after DHNB phenylthiosemicarbazone administration compared to the positive drug allopurinol.
As shown in fig. 7, different doses of DHNB dephenylthiosemicarbazide were more effective in reducing mouse serum uric acid and exhibited very significant levels compared to the allopurinol group. In addition, different doses of DHNB (dehydroepiandrosterone) thiosemicarbazone have different influences on the serum uric acid level of mice, the effect of the DHNB thiosemicarbazone is better in medium dose and high dose, and is obviously different from that of the DHNB thiosemicarbazone in low dose, but the effect of the DHNB thiosemicarbazone in medium dose and high dose is equivalent to that of the DHNB thiosemicarbazone in low dose, and the DHNB thiosemicarbazone in medium dose and high dose is not obviously different. The results show that the DHNB (dehydroepiandrosterone) thiosemicarbazone has a better effect of reducing the serum uric acid level of a mouse, has a better effect than that of a positive drug allopurinol, and the optimal dosage of the DHNB thiosemicarbazone is comprehensively evaluated.
3.2.3 Inhibition effect of DHNB (dehydroepiandrosterone) phenylthiosemicarbazone on liver XOD (x-ray activating oxygen deficiency) activity of hyperuricemia model mouse
The phosphotungstic acid method is adopted to detect the mouse liver XOD vitality of a normal saline group, a high uric acid model group, a DHNB (dehydroepiandrosterone) condensed phenyl thiosemicarbazide low dose group, a DHNB condensed phenyl thiosemicarbazide medium dose group, a DHNB condensed phenyl thiosemicarbazide high dose group and an allopurinol group, as shown in figure 8, the vitality of the model group XOD is enhanced compared with that of a normal saline control group, and the success of experimental modeling is indicated. After modeling, the DHNB (dehydroepiandrosterone) thiosemicarbazone and the positive allopurinol can inhibit the activity of mouse liver XOD (X oxygen deficiency). In addition, DHNB phenylthiosemicarbazone inhibited XOD slightly more strongly than allopurinol, but there was no significant difference between the two. The results show that the DHNB phenylthiosemicarbazone has a good inhibition effect on the mouse liver XOD activity.
Toxicity study of DHNB Phenylthiosemicarbazone on animals
4.1 acute poisoning test in mice
24 healthy Kunming mice (32 +/-2 g) with half male and female are selected, divided into two groups, a DHNB (dehydroepiandrosterone) thiosemicarbazone group and an allopurinol group, wherein each group comprises 12 mice with half male and female, the mice are subjected to intragastric administration (ig) once according to the weight of 500mg/kg, and the behavior and death condition of the mice are observed and recorded after the administration. As a result, the DHNB phenylthiosemicarbazone group administered in one week had no toxic reaction, no death, and good status; and 12 mice in allopurinol group died 6 mice, wherein 4 female mice and 2 male mice. Preliminary toxicity tests indicate that DHNB phenylthiosemicarbazone is either very low in toxicity or non-toxic.
The above embodiments are merely preferred embodiments of the present invention, and any simple modifications, alterations and substitutions made to the above embodiments according to the technical essence of the present invention are within the scope of the technical solution of the present invention.
Claims (6)
1. A chemical synthetic drug DHNB (dehydroepiandrosterone) thiosemicarbazone is characterized by having the following structure:
the DHNB (dehydroepiandrosterone) condensed phenylthiosemicarbazide is prepared by taking 3, 4-dihydroxy-5-nitrobenzaldehyde (DHNB) shown in a structural formula (I) as a raw material, and carrying out condensation reaction on the DHNB condensed phenylthiosemicarbazide shown in a structural formula (II) in a solvent methanol to synthesize the DHNB condensed phenylthiosemicarbazide shown in a structural formula (III), wherein the molar ratio of the 3, 4-dihydroxy-5-nitrobenzaldehyde (DHNB) shown in the structural formula (I) to the 4-phenylthiosemicarbazide shown in the structural formula (II) is 1.0-1.05; the reaction formula of the above process is shown below:
2. the use of the chemical synthetic drug DHNB phenyl thiosemicarbazone of claim 1 in the preparation of drugs, health foods or feeds for the prevention and treatment of gout and hyperuricemia.
3. The use according to claim 2, wherein the DHNB phenylthiosemicarbazone is formulated as a pharmaceutically active ingredient in a tablet, capsule, granule, injection, liposome or sustained release controlled release formulation.
4. The use as claimed in claim 2, characterized by the use in combination with allopurinol, febuxostat or colchicine.
5. Use according to claim 2, wherein the chelation with a metal ion forms the corresponding metal complex.
6. Use according to claim 2, wherein the DHNB phenylthiosemicarbazone is used as an additive in health food or feed.
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| WO2015131200A1 (en) * | 2014-02-28 | 2015-09-03 | Baylor College Of Medicine | Small molecule xanthine oxidase inhibitors and methods of use |
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| CN1582284A (en) * | 2001-11-07 | 2005-02-16 | 霍夫曼-拉罗奇有限公司 | Aminopyrimidines and pyridines |
| WO2015131200A1 (en) * | 2014-02-28 | 2015-09-03 | Baylor College Of Medicine | Small molecule xanthine oxidase inhibitors and methods of use |
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| Investigation of the interaction between benzaldehyde thiosemicarbazone compounds and xanthine oxidase;Mengrong Li 等;《Journal of Molecular Structure》;20180202;第1159卷;第26页Table 1，第24页2.2. Synthesis of compounds和Fig 1 * |
| Mengrong Li 等.Investigation of the interaction between benzaldehyde thiosemicarbazone compounds and xanthine oxidase.《Journal of Molecular Structure》.2018,第1159卷第23-32页. * |
| Synthesis, crystal structures, and biological activities of 2-thiophene N(4)-methylthiosemicarbazone and its unusual hexanuclear silver(I) cluster;Ming-Xue Li 等;《Inorganic Chemistry Communications》;20100715;第13卷;第1269页Fig 1和第1271页References 25 * |
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