Application of a gold preparation in preparing a drug for treating feline infectious peritonitis

Abstract

The present invention relates to the use of a gold preparation in the preparation of a drug for treating feline infectious peritonitis. The gold preparation provided by the present invention comprises gold nanoclusters, gold nanoparticles, auranofin molecules, and gold thioglucosinolate molecules. This invention, for the first time, discovers and confirms that the provided gold preparation can inhibit inflammation caused by feline infectious peritonitis (FIP), effectively control FIP progression, and significantly improve the survival rate and quality of life of affected animals. This provides a low-cost, high-potential new FIP treatment strategy for animal clinics, and pioneeringly fills a gap in the application of gold preparations in the veterinary treatment of FIP.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to application of a gold preparation in preparing a medicine for treating feline infectious peritonitis. Background Art

[0002] There is currently no widely approved effective treatment for feline infectious peritonitis (FIP). Existing treatment options include: 1. Broad-spectrum medications, such as nucleoside analogs, have shown some efficacy in some clinical cases. However, these drugs are expensive and inconvenient to administer, often requiring long-term injections, and not all cases are effective. Furthermore, these medications have limited efficacy in controlling the severe vasculitis and granulomatous inflammation associated with FIP, making them insufficiently effective for the comprehensive treatment of FIP. 2. Immunosuppressants, such as cyclosporine and prednisolone, can alleviate the inflammatory symptoms of FIP to some extent. However, long-term use of these drugs can cause significant side effects, such as immunosuppression and renal damage, significantly limiting their use in FIP treatment. 3. Supportive care, including fluid replacement and nutritional support, can alleviate symptoms in the short term, but their effectiveness in FIP treatment is limited and they do not address the underlying cause.

[0003] In addition, gold preparations, particularly auranofin, have been used in human medicine to treat rheumatoid arthritis. However, there are currently no documents or patents that disclose or suggest that gold preparations, particularly in the form of gold nanoclusters, gold nanoparticles, or gold thioglucosinolate, can effectively treat FIP.

[0004] Therefore, there is an urgent need to develop a new drug for the treatment of FIP to inhibit the inflammation caused by FIP and meet the needs of clinical treatment. Summary of the Invention

[0005] In order to solve the above technical problems, the present invention provides an application of a gold preparation in the preparation of a drug for feline infectious peritonitis.

[0006] In a first aspect, the present invention provides a use of a gold preparation in the preparation of a drug for treating feline infectious peritonitis, wherein the gold preparation comprises one or more of gold nanoclusters, gold nanoparticles, auranofin molecules, and thioglucosinolate gold molecules.

[0007] In some embodiments of the present invention, the chemical composition of the gold nanoclusters is Au x P y , x=an integer of 3-200, y=an integer of 2-220, and P represents a peptide and / or protein molecule.

[0008] In some embodiments of the present invention, x is an integer of 9-144, such as 9, 18, 22, 25, 29, 38, 102, 144, etc.

[0009] In some embodiments of the present invention, y is an integer of 8-60, such as 8, 14, 16, 18, 24, 27, 44, 60, etc.

[0010] In some embodiments of the present invention, the peptide and / or protein molecule is a peptide and / or protein molecule containing one or more of sulfhydryl, amine, imidazole, carboxyl and phenolic hydroxyl groups.

[0011] In some embodiments of the present invention, the peptide and / or protein molecule is a peptide molecule and / or protein molecule containing cysteine.

[0012] In some embodiments of the present invention, the gold nanoparticles are gold nanoparticles modified with peptides and / or protein molecules containing one or more of thiol, amine, imidazole, carboxyl and phenolic hydroxyl groups, or gold nanoparticles modified with citric acid.

[0013] In some embodiments of the present invention, the diameter of the gold nanoparticles is 5-100 nm.

[0014] In a second aspect, the present invention provides a drug for treating feline infectious peritonitis, wherein the drug comprises a gold preparation;

[0015] The gold preparation has the same meaning as the gold preparation described in the first aspect.

[0016] In some embodiments of the present invention, the components of the drug further include pharmaceutically acceptable excipients, and the excipients include one or more of fillers, stabilizers, and delivery vehicles.

[0017] In some embodiments of the present invention, the dosage form of the drug is an injection dosage form, an oral dosage form, a respiratory atomization dosage form or a transdermal dosage form.

[0018] The technical solution provided by the embodiment of the present invention has the following advantages compared with the existing technology:

[0019] This invention discovered and confirmed for the first time that specific gold preparations (gold nanoclusters, gold nanoparticles, auranofin, and thioglucosinolate gold) can inhibit the inflammation caused by FIP, effectively control the progression of FIP, and significantly improve the survival rate and quality of life of diseased animals. It provides a low-cost, high-potential new strategy for the treatment of FIP in animal clinics, and pioneeringly fills the gap in the application of gold preparations in the field of FIP treatment in veterinary medicine.

[0020] The various gold preparation forms provided by the present invention provide broad space and flexibility for the subsequent development of different dosage forms. The drugs provided by the present invention have a variety of dosage forms to choose from, and the most appropriate route of administration can be selected according to the condition of the patient, thereby improving the convenience and effectiveness of treatment. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the invention and, together with the description, serve to explain the principles of the invention.

[0022] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, for ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative labor.

[0023] Figure 1 This is a curve of body temperature changes of cats treated with FIP using gold nanoclusters as described in Example 1 of the present invention;

[0024] Figure 2 This is a graph showing weight changes in cats treated with FIP using the gold nanoclusters described in Example 1 of the present invention;

[0025] Figure 3 This is a graph showing the clinical symptom scores of cats treated with FIP using gold nanoclusters as described in Example 1 of the present invention;

[0026] Figure 4 This is a graph showing the survival of cats treated with FIP using gold nanoclusters as described in Example 1 of the present invention;

[0027] Figure 5 This is a graph showing the viral load test results of the peripheral blood of cats treated with FIP using the gold nanoclusters described in Example 1 of the present invention;

[0028] Figure 6 This is a curve of body temperature changes in cats treated with FIP using glucosinolate gold as described in Example 2 of the present invention;

[0029] Figure 7 This is a graph showing weight changes in cats treated with FIP gold glucosinolate according to Example 2 of the present invention;

[0030] Figure 8 This is a clinical symptom scoring chart for cats treated with FIP using glucosinolate gold as described in Example 2 of the present invention;

[0031] Figure 9 This is a graph showing the survival of cats treated with FIP using glucosinolate gold as described in Example 2 of the present invention;

[0032] Figure 10 This is a graph showing the results of peripheral blood viral load testing of cats with FIP treated with glucosinolate gold as described in Example 2 of the present invention. DETAILED DESCRIPTION

[0033] In order to more clearly understand the above-mentioned objectives, features and advantages of the present invention, the scheme of the present invention will be further described below. It should be noted that, in the absence of conflict, the embodiments of the present invention and the features therein can be combined with each other.

[0034] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein; it is obvious that the embodiments in the specification are only part of the embodiments of the present invention, rather than all the embodiments.

[0035] The present invention provides an application of a gold preparation in preparing a medicine for treating feline infectious peritonitis. The gold preparation comprises one or more of gold nanoclusters, gold nanoparticles, auranofin molecules and gold thioglucosinolate molecules.

[0036] In the present invention, auranofin has a long history of oral use and relatively rich safety data, which provides an important reference basis and safety starting point for veterinary development.

[0037] The gold preparations (especially gold nanoclusters / particles) of the present invention can be modified by surface ligands (such as peptides / proteins, etc.) to further optimize their in vivo distribution, metabolic pathways (pharmacokinetics) and safety characteristics.

[0038] In some embodiments of the present invention, the chemical composition of the gold nanoclusters is Au x P y , x=an integer of 3-200, y=an integer of 2-220, and P represents a peptide and / or protein molecule.

[0039] In some embodiments of the present invention, x==an integer from 9 to 144, such as 9, 18, 22, 25, 29, 38, 102, 144, etc.

[0040] In some embodiments of the present invention, y is an integer of 8-60, such as 8, 14, 16, 18, 24, 27, 44, 60, etc.

[0041] In some embodiments of the present invention, the peptide and / or protein molecule is a peptide and / or protein molecule containing one or more of a sulfhydryl group, an amine group, an imidazole group, a carboxyl group, and a phenolic hydroxyl group. The groups selected in the present invention can bind to the surface of the gold cluster and coat the surface of the gold cluster to stabilize the gold cluster.

[0042] In some embodiments of the present invention, the gold nanoclusters include Au 18 P 14 , Au 29 P 27 , Au 38 P 27 .

[0043] In some embodiments of the present invention, the peptide and / or protein molecule is a peptide molecule and / or protein molecule containing cysteine.

[0044] In some embodiments of the present invention, the gold nanoparticles are gold nanoparticles modified with peptides and / or protein molecules containing one or more of thiol, amine, imidazole, carboxyl and phenolic hydroxyl groups, or gold nanoparticles modified with citric acid.

[0045] In some embodiments of the present invention, the diameter of the gold nanoparticles is 5-100 nm, for example, 5 nm, 25 nm, 45 nm, 65 nm, 85 nm, 100 nm, etc.

[0046] In a second aspect, the present invention provides a drug for treating feline infectious peritonitis, wherein the drug comprises a gold preparation;

[0047] The gold preparation has the same meaning as the gold preparation described in the first aspect.

[0048] In some embodiments of the present invention, the components of the drug further include pharmaceutically acceptable excipients, and the excipients include one or more of fillers, stabilizers, and delivery vehicles.

[0049] In some embodiments of the present invention, the dosage form of the drug is an injection dosage form, an oral dosage form, a respiratory atomization dosage form or a transdermal dosage form.

[0050] The drug of the present invention can be prepared into a variety of dosage forms, and the most appropriate route of administration can be selected according to the specific conditions of the diseased animal (such as disease stage and medication compliance), thereby improving the convenience and effectiveness of treatment.

[0051] The glucosinolate gold in the embodiment of the present invention is obtained from conventional commercially available gold nanoclusters Au 29 P 27 (GA) is derived from artificial synthesis, gold nanoclusters Au 29 P 27 (GA) Synthesis method reference [F.Gao, Q.Yuan, P.Cai, L.Gao, L.Zhao, M.Liu, Y.Yao, Z.Chai, X.Gao, Au clusters treat rheumatoidarthritis with uniquely reversing cartilage / bone destruction, Adv.Sci.6 ,1801671(2019)].

[0052] Example 1

[0053] Evaluation of the efficacy of gold nanoclusters in treating feline infectious peritonitis

[0054] Preparation of gold nanoclusters:

[0055] Reduced glutathione and chloroauric acid were dissolved in deionized water at a molar ratio of 3:2, and the mixture was rapidly stirred for 10 minutes. Then, the mixture was continuously stirred and reacted at 70° C. for 24 hours in the dark to obtain the gold nanoclusters.

[0056] The test process is as follows:

[0057] Eight British shorthair cats were randomly divided into a positive control group and a treatment group.

[0058] (1) Virus challenge: Feline infectious peritonitis virus (FIPV) was inoculated orally and nasally with a dose of 1×10^5 TCID 50 .

[0059] (2) Clinical monitoring: The weight, temperature, and clinical status scores of all animals were monitored and recorded daily. The clinical status scores were based on observations of the following symptoms: survival status (death / dying), mental state, physical appearance (such as fur, eye and nasal discharge), behavioral activity level, excreta characteristics (feces), and appetite / drinking willingness.

[0060] (3) Dosing (as the first day): When the animals showed obvious clinical symptoms, the treatment group began to receive subcutaneous injection of gold at a dose of 8 mg / kg. The positive control group received an equal volume of normal saline at the same time.

[0061] (4) Treatment period: Subcutaneous injection once daily for 4 weeks. During this period, body weight, temperature, clinical status score and survival status were monitored daily.

[0062] (5) End point: The end point of the trial is the end of treatment (i.e., drug administration starts two weeks after infection and lasts for 4 weeks).

[0063] (6) Blood sampling and testing: Blood samples were collected once before the challenge and once a week after the start of drug administration. The TCID 50 ) to assess viral load.

[0064] The test results are as follows Figures 1 to 6 As shown:

[0065] like Figure 1 As shown in the figure, the body temperature of cats in both the positive control group and the treatment group remained stable within two weeks of administration. After two weeks of administration, the body temperature of cats in the positive control group showed an increasing trend, showing a persistent high fever; while the fever symptoms in the treatment group were effectively suppressed after administration.

[0066] like Figure 2As shown in the figure, the weight of cats in the positive control group and the treatment group remained stable within 2 weeks after administration. After 2 weeks of administration, the weight of cats in the positive control group decreased; in contrast, the weight of cats in the treatment group increased slightly.

[0067] like Figure 3 As shown in the figure, the clinical status scores of cats in each group decreased slightly within 2 weeks after administration. After 2 weeks of administration, the clinical status of cats in the positive control group deteriorated rapidly, while the clinical status of cats in the treatment group gradually improved and maintained a stable level.

[0068] like Figure 4 As shown in the figure, cats in the positive control group began to die on the 18th day after administration of the drug to the treatment group, and died successively on the 21st, 25th, and 26th days. No cats in the treatment group died during the termination period of the trial (28th day after administration).

[0069] like Figure 5 As shown in the figure, the viral load in the peripheral blood of cats in the positive control group and the treatment group increased significantly on the 7th and 14th days after administration. After administration, the viral load in the treatment group further decreased to a lower level on the 21st and 28th days.

[0070] The above results show that the gold nanoclusters provided by the present invention can significantly inhibit feline infectious peritonitis.

[0071] Example 2

[0072] Evaluation of the efficacy of glucosinolate gold in the treatment of feline infectious peritonitis

[0073] In this example, the drug is commercially available thioglucosinolate gold. Eight British shorthair cats were randomly divided into a positive control group and a treatment group. The experimental process is as follows:

[0074] (1) Virus challenge: Feline infectious peritonitis virus (FIPV) was inoculated orally and nasally with a dose of 1×10^5 TCID 50 .

[0075] (2) Clinical monitoring: The weight, temperature, and clinical status scores of all animals were monitored and recorded daily. The clinical status scores were based on observations of the following symptoms: survival status (death / dying), mental state, physical appearance (such as fur, eye and nasal discharge), behavioral activity level, excreta characteristics (feces), and appetite / drinking willingness.

[0076] (3) Dosing (as Day 1): When the animals showed obvious clinical symptoms, the treatment group began to receive subcutaneous injection of gold at a dose of 4 mg / kg. The positive control group received an equal volume of normal saline at the same time.

[0077] (4) Treatment period: Subcutaneous injection once daily for 4 weeks. During this period, body weight, temperature, clinical status score and survival status were monitored daily.

[0078] (5) End point: The end point of the trial is the end of treatment (i.e., drug administration starts two weeks after infection and lasts for 4 weeks).

[0079] (6) Blood sampling and testing: Blood samples were collected once before the infection and once a week after the group was enrolled and administered. Blood biochemical indicators were tested and the TCID 50 ) to assess viral load.

[0080] The test results are as follows Figures 6 to 10 As shown:

[0081] like Figure 6 As shown in the figure, the body temperature of cats in both the positive control group and the treatment group remained stable within two weeks of administration. After two weeks of administration, the body temperature of cats in the positive control group showed an increasing trend, showing a persistent high fever; while the fever symptoms in the treatment group were effectively suppressed after administration.

[0082] like Figure 7 As shown in the figure, the weight of cats in the positive control group and the treatment group remained stable within 2 weeks after administration. After 2 weeks of administration, the weight of cats in the positive control group decreased; in contrast, the weight of cats in the treatment group increased slightly.

[0083] like Figure 8 As shown in the figure, the clinical status scores of cats in each group decreased slightly within 2 weeks after administration. After 2 weeks of administration, the clinical status of cats in the positive control group deteriorated rapidly, while the clinical status of cats in the treatment group gradually improved and maintained a stable level.

[0084] like Figure 9 As shown in the figure, cats in the positive control group began to die on the 19th day after administration of the drug to the treatment group, and died successively on the 21st, 24th and 25th days. No cats in the treatment group died during the termination period of the trial (28th day after administration).

[0085] like Figure 10 As shown in the figure, the viral load in the peripheral blood of cats in both the positive control group and the treatment group increased significantly on days 7 and 14 after administration. On days 21 and 28 after administration, the viral load in the treatment group gradually decreased. These results indicate that glucosinolate gold can significantly inhibit feline infectious peritonitis.

[0086] It should be noted that, in this document, relational terms such as "first" and "second" are used only to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship or order between these entities or operations. Moreover, the terms "comprises," "comprising," or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article, or device comprising a series of elements includes not only those elements, but also other elements not explicitly listed, or elements inherent to such process, method, article, or device. In the absence of further limitations, an element defined by the phrase "comprising a ..." does not exclude the presence of other identical elements in the process, method, article, or device comprising the element.

[0087] The foregoing description is intended only to provide specific embodiments of the present invention, which will enable those skilled in the art to understand and implement the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not intended to be limited to the embodiments described herein, but is to be construed in the widest manner consistent with the principles and novel features disclosed herein.

Claims

1. Use of a gold preparation in the preparation of a medicament for treating feline infectious peritonitis, characterized in that: The gold preparation is gold nanoclusters Au 29 SG 27 and one or both of the gold glucosinolate molecules; The preparation of the gold nanoclusters includes the following steps: dissolving reduced glutathione and chloroauric acid in deionized water at a molar ratio of 3:2, rapidly stirring and mixing for 10 minutes, and then continuously stirring and reacting at 70° C. for 24 hours under light-proof conditions to obtain the gold nanoclusters.

Images