Use of compound npb-1575 or a derivative thereof for the preparation of a medicament for the treatment of parkinson's disease

Abstract

The application belongs to the technical field of medicines, and discloses application of a compound NPB-1575 in preparation of a medicine for preventing or treating Parkinson's disease. + The compound NPB-1575 can significantly improve the cell survival rate of a rotenone, 6-OHDA and MPP + The application provides a treatment drug with definite curative effect for preventing or delaying the progress of Parkinson's disease, and is also a new indication of the compound NPB-1575, and has a good application prospect.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical use, and more specifically to the use of compound NPB-1575 or its derivatives in the preparation of medicaments for the treatment or prevention of Parkinson's disease. Background Technology

[0002] Parkinson's disease (PD) is the second most common neurodegenerative disease worldwide, affecting approximately 2% of the population aged 65 and older. Clinical manifestations of PD include resting tremor, rigidity, bradykinesia, and ataxia, accompanied by non-motor symptoms such as depression, constipation, and olfactory dysfunction. Its characteristic pathological change is the degeneration and loss of dopaminergic (DA) neurons in the substantia nigra pars compacta. Epidemiological studies indicate that most PD cases are sporadic, with 5%-10% being familial. The etiology of PD remains unclear, but it is believed to be related to genetic factors, environmental toxins, and aging. Theories regarding the pathogenesis of PD mainly include neuroinflammation, mitochondrial dysfunction, oxidative stress, calcium overload, excitatory amino acid toxicity, and apoptosis.

[0003] Parkinson's disease (PD) treatment includes both pharmacological and non-pharmacological therapies. Pharmacological therapy remains a crucial treatment method for PD management. Based on their mechanisms of action, anti-Parkinson's drugs are classified into six categories: dopaminergic drugs, dopamine receptor (DR) agonists, anticholinergic drugs, amantadine, monoamine oxidase inhibitors, and catechol-O-methyltransferase (COMT) inhibitors. However, these drugs have limited efficacy and cannot slow disease progression, cure the disease, or reverse its neurodegenerative effects. Levodopa is one of the most commonly used drugs for treating Parkinson's disease clinically, but it cannot stop the natural progression of the disease, and its efficacy gradually decreases with long-term use, accompanied by serious side effects. Therefore, neuroprotective therapy to slow the degenerative process of PD and the search for drugs that can reduce the loss of dopaminergic neurons and block or delay disease progression are critical issues that urgently need to be addressed.

[0004] Compound NPB-1575 is an isopentenyl bibenzyl compound, first isolated from the plant licorice. The applicant and their team constructed a high-yield *Escherichia coli* strain producing the bibenzyl skeleton product using a highly active bibenzyl synthase isolated from *Dendrobium officinale*. Simultaneously, utilizing highly active and substrate-generalizable methyltransferases, isopentenyltransferases, and glycosyltransferases, along with their corresponding donor biosynthetic modules, they constructed a highly efficient post-modification modular strain. Through co-culture engineering, they synthesized the isopentenyl bibenzyl compound NPB-1575 (Structural diversification of bioactive bibenzyls through modular co-culture leading to the discovery of a novel neuroprotective agent. *Acta Pharmaceutica Sinica B*, 2023, 13, 1771-1785.).

[0005] Compound NPB-1575 possesses the functions of scavenging free radicals in vitro, reducing oxidative stress damage, and improving neuroinflammation. It can block multiple pathological processes of brain injury caused by ischemic stroke, exhibiting strong anti-ischemic and neuroprotective effects. To date, no research has been reported on the use of NPB-1575 and its derivatives for the treatment or prevention of Parkinson's disease.

[0006] Therefore, the main objective of this invention is to utilize rotenone, 6-OHDA, and MPP. + By using a damaged PC12 cell model, the study investigated the ameliorative effect of compound NPB-1575 on the aforementioned Parkinson's disease cell model, providing a potential drug for the prevention and treatment of Parkinson's disease. Summary of the Invention

[0007] The technical problem solved by this invention is the application of compound NPB-1575 or its derivatives in the preparation of drugs for the prevention or treatment of Parkinson's disease.

[0008] To address the technical problem of this invention, the present invention provides the following technical solution:

[0009] The first aspect of the present invention is to provide the use of isopentenyl bibenzyl derivatives having the structure shown in general formula 1 or pharmaceutically acceptable salts thereof in the preparation of drugs for the prevention or treatment of Parkinson's disease.

[0010]

[0011] In this case, the substituents R1 and R2 are each independently selected from hydrogen or hydroxyl groups.

[0012]

[0013] As a preferred embodiment, the structural formula of compound NPB-1575 is as follows. The compound is selected from 2-isopentenyl-3,4′,5-trihydroxybibenzyl, 2-isopentenyl-3,3′,5-trihydroxybibenzyl, and 2-isopentenyl-3,3′,4′,5-tetrahydroxybibenzyl. The pharmaceutically acceptable salt is selected from organic or inorganic acid salts, including hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, malate, fumarate, tartrate, methanesulfonate, carbonate, oxalate, lactate, succinate, or gluconate.

[0014] A second aspect of the present invention is to provide an application of a pharmaceutical composition in the preparation of a drug for the prevention or treatment of Parkinson's disease. The pharmaceutical composition comprises the isopentenyl bibenzyl derivative described in the first aspect, its pharmaceutically acceptable salt, and a pharmaceutically acceptable carrier. The pharmaceutically acceptable carrier is selected from lipid carriers, polymer carriers, protein and polypeptide carriers, inorganic nanocarriers, carbon-based nanomaterials, hydrogels, and biodegradable materials.

[0015] The compounds of this invention or pharmaceutical compositions containing them can be administered in unit dose form via enteral or non-enteric routes, such as oral, intravenous, intramuscular, subcutaneous, nasal, oral mucosa, eye, lung and respiratory tract, skin, vagina, rectum, etc.

[0016] Dosage forms can be liquid, solid, or semi-solid. Liquid dosage forms can include solutions (including true solutions and colloidal solutions), emulsions (including o / w, w / o, and double emulsions), suspensions, injections (including aqueous injections, powder injections, and infusions), eye drops, nasal drops, lotions, and liniments, etc.; solid dosage forms can include tablets (including regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets), capsules (including hard capsules, soft capsules, and enteric-coated capsules), granules, powders, microcapsules, pellets, suppositories, films, patches, aerosols, and sprays, etc.; semi-solid dosage forms can include ointments, gels, and pastes, etc.

[0017] The compounds of this invention can be formulated into conventional formulations, sustained-release formulations, controlled-release formulations, targeted formulations, and various microparticle delivery systems.

[0018] To formulate the compounds of the present invention into tablets, a wide variety of excipients known in the art can be used, including diluents, binders, wetting agents, disintegrants, lubricants, and flow aids. Diluents can be starch, dextrin, sucrose, glucose, lactose, mannose, sorbitol, xylitol, microcrystalline cellulose, calcium sulfate, dicalcium phosphate, calcium carbonate, etc.; wetting agents can be water, ethanol, isopropanol, etc.; binders can be starch paste, dextrin, syrup, honey, glucose solution, microcrystalline cellulose, gum arabic paste, gelatin paste, sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, ethyl cellulose, acrylic resin, carbomer, polyvinylpyrrolidone, polyethylene glycol, etc.; disintegrants can be dry starch, microcrystalline cellulose, low-substituted hydroxypropyl cellulose, cross-linked polyvinylpyrrolidone, cross-linked sodium carboxymethyl cellulose, sodium carboxymethyl starch, sodium bicarbonate and citric acid, polyoxyethylene sorbitol fatty acid ester, sodium dodecyl sulfonate, etc.; lubricants and flow aids can be talc, silica, stearate, tartaric acid, liquid paraffin, polyethylene glycol, etc.

[0019] Tablets can also be further processed into coated tablets, such as sugar-coated tablets, film-coated tablets, enteric-coated tablets, or bilayer and multilayer tablets.

[0020] To formulate the drug delivery unit into capsules, the active ingredient, the compound of the present invention, can be mixed with a diluent and a disintegrant, and the mixture can be placed directly into hard or soft capsules. Alternatively, the active ingredient, the compound of the present invention, can be first formed into granules or microspheres with a diluent, binder, and disintegrant, and then placed into hard or soft capsules. Various diluents, binders, wetting agents, disintegrants, and disintegrants used to prepare tablets of the compound of the present invention can also be used to prepare capsules of the compound of the present invention.

[0021] To prepare the compounds of this invention into injectable formulations, water, ethanol, isopropanol, propylene glycol, or mixtures thereof can be used as solvents, and appropriate amounts of commonly used solubilizers, co-solvents, pH adjusters, and osmotic pressure adjusters can be added. Solubilizers or co-solvents can be poloxamer, lecithin, hydroxypropyl-β-cyclodextrin, etc. pH adjusters can be phosphates, acetates, hydrochloric acid, sodium hydroxide, etc.; osmotic pressure adjusters can be sodium chloride, mannitol, glucose, phosphates, acetates, etc. If preparing lyophilized powder injections, mannitol, glucose, etc., can also be added as supporting agents.

[0022] In addition, colorants, preservatives, flavorings, tasters or other additives may be added to pharmaceutical preparations if necessary.

[0023] To achieve the purpose of medication and enhance the therapeutic effect, the drug or drug composition of the present invention can be administered using any known method of administration.

[0024] The compounds or compositions of the present invention can be taken alone or in combination with other therapeutic or symptomatic drugs. When the compounds of the present invention have a synergistic effect with other therapeutic drugs, their dosage should be adjusted according to the actual situation.

[0025] The compounds of this invention can be obtained by biosynthesis, chemical synthesis, or bio-chemical synthesis.

[0026] Beneficial technical effects:

[0027] Using rotenone, 6-OHDA and MPP + Experiments were conducted using a PC12 cell injury model to evaluate the neuroprotective effect of compound NPB-1575 in Parkinson's disease. MTT results further indicated that compound NPB-1575 significantly increased rotenone, 6-OHDA, and MPP levels. + The increased cell survival rate in the damaged PC12 cell model indicates a neuroprotective effect. This invention provides a clinically effective treatment drug that can halt or delay the progression of Parkinson's disease. It also represents a novel indication for compound NPB-1575 beyond ischemic stroke, demonstrating promising application prospects. Attached Figure Description

[0028] Figure 1 NPB-1575 improved cell survival in a rotenone-damaged PC12 cell model. A. The effect of 0.1–3 μM NPB-1575 on the survival of rotenone-damaged PC12 cells was detected by MTT assay. # indicates that compared with the control group, [the percentage of NPB-1575 decreased]. ### P<0.001, * indicates comparison with the rotenone group, where *P<0.05.

[0029] Figure 2 NPB-1575 improved cell survival in a 6-OHDA-damaged PC12 cell model. A. The effect of 0.1–10 μM NPB-1575 on the survival of 6-OHDA-damaged PC12 cells was detected by MTT assay. # indicates that compared with the control group, [the percentage of NPB-1575 decreased]. ### P<0.001, * indicates comparison with the 6-OHDA group, where **P<0.01.

[0030] Figure 3 NPB-1575 improves MPP + Cell survival in a PC12 cell model with damage. A. MTT assay to detect the effect of 0.1-10 μM NPB-1575 on MPP. + The effect of damage on PC12 cell survival. # indicates the difference compared to the control group. ### P < 0.001, * indicates P < 0.001. + Compared with the previous group, *P<0.05. Detailed Implementation

[0031] To further understand the present invention, the following embodiments are only used to further illustrate the present invention, but do not imply any limitation on the present invention.

[0032] Example 1. NPB-1575 significantly improves cell survival in a rotenone-damaged PC12 cell model.

[0033] 1. Cells and Compounds

[0034] Cell lines:

[0035] PC12 cell line was purchased from Wuhan Pronosei Life Science Technology Co., Ltd.

[0036] Compounds:

[0037] Rotenone (CAS: 83-79-4) was purchased from Sigma Reagents.

[0038] 2. Methods

[0039] 2.1 MTT assay for cell viability

[0040] PC12 cells were used at a rate of 1*10 6 Cells were cultured at a density of [number] cells / ml in 96-well plates, with 100 μL of cell suspension added to each well. After complete cell adhesion, 10 μL of 5 μM rotenone was added to each well of the model group, bringing the final concentration to 0.5 μM. In addition to rotenone, the drug treatment groups received 1 μM, 3 μM, 10 μM, and 30 μM NPB-1575 solutions, respectively, bringing the final concentrations to 0.1 μM, 0.3 μM, 1 μM, and 3 μM. After thorough mixing of all reagents in each well, the 96-well plates were incubated in a cell culture incubator. After 48 hours, 10 μL of 5 mg / ml MTT solution was added to each well, and the 96-well plates were incubated for another 4 hours. Four hours later, the liquid in each well of the 96-well plate was discarded, and 150 μL of DMSO was added to each well. After thorough mixing, the absorbance of each well was measured at 570 nm, and the cell viability of each group was calculated.

[0041] 2.2 Data Statistical Analysis

[0042] Data are presented as mean ± standard error (mean ± SEM). Statistical analysis was performed using Student's t-test. # indicates a difference between the rotenone group and the control group. ### P<0.001* indicates that the NPB-1575 group was compared with the rotenone group, where *P<0.05.

[0043] 3. Results

[0044] 3.1 NPB-1575 significantly improved cell survival in a rotenone-damaged PC12 cell model.

[0045] The results are shown in Table 1 and Figure 1 As shown, NPB-1575 significantly improved the survival rate of PC12 cells damaged by rotenone by 19.01%, suggesting that NPB-1575 has an ameliorative effect on rotenone-induced damage to PC12 cells.

[0046] Table 1. Effect of NPB-1575 on cell viability of PC12 cells damaged by rotenone (n=4)

[0047]

[0048] Example 2. NPB-1575 significantly improves cell survival in a 6-OHDA-damaged PC12 cell model. 1. Cells and Compounds

[0049] Cell lines:

[0050] PC12 cell line was purchased from Wuhan Pronosei Life Science Technology Co., Ltd.

[0051] Compounds:

[0052] 6-OHDA (CAS: 28094-15-7) was purchased from Maclean's Reagent Company.

[0053] 2. Methods

[0054] 2.1 MTT assay for cell viability

[0055] PC12 cells were used at a rate of 1*10 6 Cells were cultured at a density of [number] cells / ml in 96-well plates, with 100 μL of cell suspension added to each well. After complete cell adhesion, 10 μL of 750 μM 6-OHDA was added to each well of the model group, bringing the final concentration to 75 μM. In addition to the above 6-OHDA, 1 μM, 10 μM, and 100 μM NPB-1575 were added to each well, bringing the final concentrations to 0.1 μM, 1 μM, and 10 μM, respectively. After thorough mixing of the reagents in each well, the 96-well plates were incubated in a cell culture incubator. After 24 hours, 10 μL of 5 mg / ml MTT solution was added to each well, and the 96-well plates were incubated for another 4 hours. Four hours later, the liquid in each well of the 96-well plate was discarded, and 150 μL of DMSO was added to each well. After thorough mixing, the absorbance of each well was measured at 570 nm, and the cell viability of each group was calculated.

[0056] 2.2 Data Statistical Analysis

[0057] Data are presented as mean ± standard error (mean ± SEM). Statistical analysis was performed using Student's t-test. # indicates a difference between the 6-OHDA group and the control group. ### P<0.001* indicates that the NPB-1575 group was compared with the 6-OHDA group, where **P<0.01.

[0058] 3. Results

[0059] 3.1 NPB-1575 can significantly improve cell survival in a 6-OHDA-damaged PC12 cell model.

[0060] The results are shown in Table 2 and Figure 2 As shown, NPB-1575 significantly improved the survival rate of 6-OHDA-damaged PC12 cells by 20.82%, suggesting that NPB-1575 has an ameliorative effect on 6-OHDA-induced PC12 cell damage.

[0061] Table 2. Effect of NPB-1575 on cell viability of PC12 cells damaged by 6-OHDA (n=3)

[0062]

[0063] Example 3. NPB-1575 can significantly improve MPP. + Cell survival in the PC12 cell model of injury

[0064] 1. Cells and Compounds

[0065] Cell lines:

[0066] PC12 cell line was purchased from Wuhan Pronosei Life Science Technology Co., Ltd.

[0067] Compounds:

[0068] MPP + (CAS: 36913-39-0) Purchased from Aladdin Reagent Company.

[0069] 2. Methods

[0070] 2.1 MTT assay for cell viability

[0071] PC12 cells were used at a rate of 1*10 6 Cells were cultured at a density of [number] cells / ml in 96-well plates, with 100 μL of cell suspension added to each well. After complete cell adhesion, 10 mM MPP was added to each well of the model group. + 10 μL was added to achieve a final concentration of 1 mM in each well. In addition to the above-mentioned MPP, the dosing group also received [the following]. +In addition, 1 μM, 10 μM, and 100 μM NPB-1575 solutions were added to each well, bringing the final concentrations to 0.1 μM, 1 μM, and 10 μM, respectively. After thorough mixing of the reagents in each well, the 96-well plate was placed in a cell culture incubator for further incubation. After 24 hours, 10 μL of 5 mg / ml MTT solution was added to each well, and the plate was incubated for another 4 hours. After 4 hours, the liquid in each well was discarded, and 150 μL of DMSO was added to each well. After thorough mixing, the absorbance of each well was measured at 570 nm, and the cell viability of each group was calculated.

[0072] 2.2 Data Statistical Analysis

[0073] Data are presented as mean ± standard error (mean ± SEM). Statistical analysis was performed using Student's t-test. # indicates MPP. + Compared with the control group, the group ### P<0.001* indicates that the NPB-1575 administration group was significantly different from the MPP group. + Compared with the previous group, *P<0.05.

[0074] 3. Results

[0075] 3.1 NPB-1575 can significantly improve MPP. + Cell survival in the PC12 cell model of injury

[0076] The results are shown in Table 3 and Figure 3 As shown, NPB-1575 can significantly improve MPP. + The survival rate of damaged PC12 cells reached 20.40%, suggesting that NPB-1575 is effective against MPP. + It has a mitigating effect on induced damage to PC12 cells.

[0077] Table 3 NPB-1575 for MPP + Effect of damage on cell survival of PC12 cells (n=3)

[0078]

[0079] The above embodiments are merely illustrative of the technical concept and features of the present invention, intended to enable those skilled in the art to understand the content of the present invention and implement it accordingly, and should not be construed as limiting the scope of protection of the present invention. All equivalent changes or modifications made according to the spirit and essence of the present invention should be covered within the scope of protection of the present invention.

Claims

1. The use of isopentenyl bibenzyl derivatives having the structure shown in Formula 1 or pharmaceutically acceptable salts thereof in the preparation of medicaments for the treatment or prevention of Parkinson's disease. in, Substituents R1 and R2 are each independently selected from hydrogen or hydroxyl groups.

2. The application according to claim 1, characterized in that, The use of compound NPB-1575 or a pharmaceutically acceptable salt thereof in the preparation of medicaments for the treatment or prevention of Parkinson's disease.

3. The application according to claim 1 or 2, characterized in that, The Parkinson's disease mentioned is primary Parkinson's disease.

4. The application according to claim 1, characterized in that, The compound is selected from 2-isopentenyl-3,4′,5-trihydroxybibenzyl, 2-isopentenyl-3,3′,5-trihydroxybibenzyl, and 2-isopentenyl-3,3′,4′,5-tetrahydroxybibenzyl.

5. The application according to claim 1 or 2, characterized in that, The pharmaceutically acceptable salt is selected from organic or inorganic acid salts, including hydrochloride, hydrobromide, sulfate, phosphate, acetate, citrate, malate, fumarate, tartrate, methanesulfonate, carbonate, oxalate, lactate, succinate, or gluconate.

6. The use of a pharmaceutical composition in the preparation of a medicament for treating or preventing Parkinson's disease, characterized in that, Contains an effective dose of the isopentenyl bibenzyl derivative of claim 1 or a pharmaceutically acceptable salt thereof, or the compound NPB-1575 of claim 2 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier thereof.

7. The application according to claim 6, characterized in that, The pharmaceutically acceptable carriers are selected from lipid carriers, polymer carriers, protein and peptide carriers, inorganic nanocarriers, carbon-based nanomaterials, hydrogels, and biodegradable materials.

8. The application according to claim 6, characterized in that, The pharmaceutical composition is selected from solutions, emulsions, suspensions, injections, eye drops, nasal drops, lotions, and liniments; solid dosage forms may be tablets, capsules, granules, powders, microcapsules, droplets, suppositories, films, patches, aerosols, powder sprays, and sprays; semi-solid dosage forms may be ointments, gels, and pastes.

9. The application according to claim 6, characterized in that, The solutions include true solutions and colloidal solutions; the emulsions include o / w, w / o, and double emulsions; the injections include aqueous injections, powder injections, and infusions; the tablets include regular tablets, enteric-coated tablets, lozenges, dispersible tablets, chewable tablets, effervescent tablets, and orally disintegrating tablets; and the capsules include hard capsules, soft capsules, and enteric-coated capsules.

10. The application according to claim 2, characterized in that, The compound NPB-1575 or its derivatives are obtained through biosynthesis, chemical synthesis, or bio-chemical synthesis.

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