Application of Lumacaftor in treatment of Parkinson's disease

Abstract

The invention discloses an application of Lumacaftor in the treatment of Parkinson's disease. The treatment effect of Lumacaftor on PD is verified through in-vitro cell experiments and in-vivo animal experiments respectively, and it is found that after Lumacaftor is used in a microglial cell-neuron co-culture PD cell model, apoptosis pathway related protein in neurons is remarkably reduced compared with a model group; after Lumacaftor treatment is performed on a PD mouse in an MPTP mouse PD model, dopaminergic neurons of the PD mouse are remarkably increased, and the motor function is improved. The application provides a new thought for effective treatment of PD, and the application prospect is wide.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine, specifically relating to the application of Lumacaftor in the treatment of Parkinson's disease. Background Technology

[0002] Parkinson's disease (PD) is a common degenerative disease of the central nervous system. The main pathological changes are the loss of dopaminergic neurons in the substantia nigra pars compacta and the depletion of dopamine in the striatal axon terminals. Typical clinical manifestations include akinesia, rigidity, resting tremor, and postural reflex disorders.

[0003] The pathogenesis of Parkinson's disease (PD) remains unclear. Currently, approximately 50% of patients experience "wearing-off" and "on / off" phenomena within 5 years after using drugs such as levodopa. Dopamine receptor agonists may cause side effects such as drowsiness and hallucinations. These drugs only replenish dopamine but cannot stop the continuous degeneration of neurons. To date, there are no mature and effective drugs or methods to stop or reverse the progression of PD. Therefore, finding effective neuroprotective drugs for prevention and treatment has become a key focus of future in-depth research. Summary of the Invention

[0004] To overcome the shortcomings of existing technologies, this invention provides the application of Lumacaftor in the treatment of Parkinson's disease.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: The first aspect of the present invention provides the use of Lumacaftor in the preparation of a medicament for treating Parkinson's disease.

[0006] Furthermore, the Lumacaftor treats Parkinson's disease by inhibiting neuronal apoptosis, increasing dopaminergic neurons, and improving motor function through any one or more of these mechanisms.

[0007] Furthermore, the medication also includes other medications for treating Parkinson's disease.

[0008] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0009] Furthermore, the pharmaceutically acceptable excipients include diluents, binders, surfactants, humectants, adsorbents, lubricants, fillers, and disintegrants.

[0010] Furthermore, the dosage forms of the drug include gastrointestinal dosage forms and non-gastrointestinal dosage forms.

[0011] Furthermore, the gastrointestinal dosage forms include tablets, granules, capsules, solutions, dry suspensions, powders, sustained-release preparations, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0012] Furthermore, the non-gastrointestinal dosage forms include injectable dosage forms, respiratory dosage forms, cavity dosage forms, mucosal dosage forms, and skin dosage forms.

[0013] A second aspect of the invention provides the use of Lumacaftor in the preparation of drugs that inhibit neuronal apoptosis / increase dopaminergic neurons / improve motor function.

[0014] A third aspect of the invention provides a medicament for treating Parkinson's disease, the medicament comprising Lumacaftor.

[0015] Furthermore, the medication also includes other medications for treating Parkinson's disease.

[0016] Furthermore, the drug also includes pharmaceutically acceptable excipients.

[0017] Advantages and beneficial effects of the present invention: This application validated the therapeutic effect of Lumacaftor on Parkinson's disease (PD) through in vitro cell experiments and in vivo animal experiments. It was found that in a microglia-neuronal co-culture PD cell model, the use of Lumacaftor significantly reduced apoptosis-related proteins in neurons compared to the model group. In an MPTP mouse PD model, treatment with Lumacaftor significantly increased dopaminergic neurons and improved motor function. This application provides a new approach for the effective treatment of PD and has broad application prospects. Attached Figure Description

[0018] Figure 1 The graphs show the inhibition of apoptosis in SH-SY5Y cells by Lumacaftor. 1A is a Western blot plot, 1B is a p-TAK1 / TAK1 plot, 1C is a p-P65 / P65 plot, 1D is a p-IκB / IκB plot, 1E is an IL-1R1 / β-actin plot, 1F is a BAX / Bcl-2 plot, and 1G is a Cleaved Caspase3 / Caspase3 plot. Figure 2 These are immunohistochemical staining results of TH-positive nerve fibers in the striatum and TH-positive neurons in the substantia nigra of mice after Lumacaftor treatment. Among them, 2A is an immunohistochemical staining image of TH-positive nerve fibers in the striatum of mice, 2B is an immunohistochemical staining image of TH-positive neurons in the substantia nigra of mice, 2C is a statistical diagram of TH-positive nerve fibers in the striatum of mice, and 2D is a statistical diagram of TH-positive neurons in the substantia nigra of mice. Figure 3These are graphs showing how Lumacaftor improves motor function in PD mice. 3A is a graph showing the turning time of mice in the pole climbing experiment; 3B is a graph showing the total time of mice in the pole climbing experiment; 3C is a graph showing the rotational speed of mice when falling from the rotunda in the rotunda experiment; 3D is a graph showing the total distance traveled by mice in the open field experiment; 3E is a graph showing the average speed of mice in the open field experiment; 3F is a graph showing the number of times mice entered the central zone in the open field experiment; 3G is a graph showing the time mice spent in the central zone in the open field experiment; and 3H is a graph from the open field experiment. Detailed Implementation

[0019] The following provides definitions for some of the terms used in this specification. Unless otherwise stated, all technical and scientific terms used herein generally have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0020] This invention provides the application of Lumacaftor in the preparation of drugs for treating Parkinson's disease.

[0021] In some embodiments, the Lumacaftor further includes pharmaceutically acceptable salts. Pharmaceutically acceptable salts refer to any acid addition salt or base addition salt whose counterions are non-toxic to the patient at pharmaceutical doses of the salt. The main components of pharmaceutically acceptable salts are well known in the art. If pharmaceutically acceptable salts of the compounds of this application are used in these compositions, those salts are preferably derived from inorganic acids or organic acids and bases. Such acidic salts include, but are not limited to, the following: acetates, adipates, alginates, aspartates, benzoates, benzenesulfonates, hydrogen sulfates, butylates, citrates, camphorates, camphorsulfonates, cyclopentanepropionates, diglucuronides, dodecyl sulfates, ethanesulfonates, fumarates, glucoheptanoates, glycerophosphates, hemisulfates, heptanates, hexanoates, hydrochlorides, hydrobromide, hydroiodates, 2-hydroxyethanesulfonates, lactates, maleates, methanesulfonates, 2-naphthalenesulfonates. Salts, nicotinic acid salts, oxalates, pyrates, pectinates, persulfates, 3-phenyl-propionates, picrates, pentanoates, propionates, succinates, tartrates, thiocyanates, toluenesulfonates, undecanoates, hydrohalides (e.g., hydrochlorides and hydrobromates), sulfates, phosphates, nitrates, aminosulfonates, malonates, salicylates, methylene-bis-β-hydroxynaphthyl carboxylate, gentianates, hydroxyethylsulfonates, di-p-toluyl tartrate, ethanesulfonates, cyclohexylaminosulfonates, quinates, etc. Pharmaceutically acceptable base addition salts include, but are not limited to, those derived from alkali metal or alkaline earth metal bases or conventional organic bases, such as triethylamine, pyridine, piperidine, morpholine, N-methylmorpholine, ammonium salts, alkali metal salts (such as sodium and potassium salts), alkaline earth metal salts (such as calcium and magnesium salts), salts containing organic bases (such as dicyclohexylamine salts, N-methyl-D-glucosamine), and salts containing amino acids such as arginine and lysine.

[0022] The medications also include other medications for treating Parkinson's disease.

[0023] In some implementations, other medications for treating Parkinson's disease include, but are not limited to, anticholinergic drugs, amantadine, levodopa / carbidopa, dopamine receptor agonists, monoamine oxidase inhibitors, and COMT inhibitors.

[0024] The drug also includes pharmaceutically acceptable excipients.

[0025] In some embodiments, a pharmaceutically acceptable excipient is used to refer to a material that is compatible with the recipient, preferably a mammal, more preferably a human, and is suitable for delivering the active agent to the target site without terminating the activity of the agent. The toxicity or side effects (if present) associated with the pharmaceutically acceptable excipient are preferably commensurate with a reasonable risk / benefit ratio for the intended use of the active agent.

[0026] In some embodiments, pharmaceutically acceptable excipients include, but are not limited to, diluents, binders, surfactants, humectants, adsorbents, lubricants, fillers, and disintegrants. These pharmaceutically acceptable excipients are used as needed to aid in the stability of the formulation or to contribute to its activity or bioavailability, or to produce an acceptable taste or odor when taken orally. The formulation used in such a drug may be in the form of its original compound itself or optionally in the form of its pharmaceutically acceptable salt. The drug thus formulated may be administered as needed by any appropriate method known to those skilled in the art.

[0027] The diluents include, but are not limited to, lactose, sodium chloride, glucose, urea, starch, and water.

[0028] Adhesives include, but are not limited to, starch, pregelatinized starch, dextrin, maltodextrin, sucrose, gum arabic, gelatin, methylcellulose, carboxymethylcellulose, ethylcellulose, polyvinyl alcohol, polyethylene glycol, polyvinylpyrrolidone, alginate and alginates, xanthan gum, hydroxypropylcellulose and hydroxypropylmethylcellulose.

[0029] Surfactants include, but are not limited to, polyethylene oxide sorbitan fatty acid esters, sodium lauryl sulfate, glyceryl monostearate, and hexadecyl alcohol.

[0030] Humectants include, but are not limited to, glycerin.

[0031] Adsorption carriers include, but are not limited to, bentonite, silica gel, kaolin and soap clay.

[0032] Lubricants include, but are not limited to, zinc stearate, glyceryl monostearate, polyethylene glycol, talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, hydrogenated vegetable oil, sodium stearate fumarate, polyoxyethylene monostearate, monolauric sucrose ester, sodium lauryl sulfate, magnesium lauryl sulfate, and magnesium dodecyl sulfate.

[0033] Fillers include, but are not limited to, mannitol (granular or powdered), xylitol, sorbitol, maltose, erythritol, microcrystalline cellulose, polysaccharides, coupled sugars, glucose, lactose, sucrose, dextrin, starch, sodium alginate, kelp polysaccharide powder, agar powder, calcium carbonate, and sodium bicarbonate.

[0034] Disintegrants include, but are not limited to, crosylvinylpyrrolidone, sodium carboxymethyl starch, low-substituted hydroxypropylmethyl, crosylcarboxymethyl cellulose sodium, and soybean polysaccharides.

[0035] The dosage forms of the drug include gastrointestinal dosage forms and non-gastrointestinal dosage forms.

[0036] In some embodiments, the gastrointestinal dosage form includes tablets, granules, capsules, solutions, dry suspensions, powders, sustained-release formulations, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

[0037] In some embodiments, the non-gastrointestinal dosage form includes injectable dosage form, respiratory dosage form, cavity dosage form, mucosal dosage form, and skin dosage form.

[0038] In some embodiments, the injectable dosage forms include, but are not limited to, various injectables such as intravenous injections, intramuscular injections, subcutaneous injections, intradermal injections, and intracavitary injections; the respiratory dosage forms include, but are not limited to, sprays, aerosols, and powder inhalers; the cavity dosage forms include, but are not limited to, suppositories, aerosols, effervescent tablets, drops, and pills, for use in the rectum, vagina, urethra, nasal cavity, and ear canal; the mucosal dosage forms include, but are not limited to, eye drops, nasal drops, ointments, mouthwashes, sublingual tablets, adhesive tablets, and patches; and the skin dosage forms include, but are not limited to, topical solutions, lotions, liniments, ointments, plasters, pastes, and patches.

[0039] In some embodiments, the drug can be manufactured using methods well-known in the art, such as conventional granulation, mixing, dissolving, encapsulation, lyophilization, or emulsification. The drug can be formulated in various forms, including granules, precipitates or microparticles, powders (including lyophilized powders, rotary-dried powders, or spray-dried powders, amorphous powders), tablets, capsules, syrups, suppositories, injections, emulsions, elixirs, suspensions, or solutions.

[0040] In some embodiments, the drug is formulated for administration to mammals. The drug can be administered via various routes, including intravenous injection, intraperitoneal injection, subcutaneous injection, intramuscular injection, oral administration, via mucosa, rectum, transdermal, or inhalation.

[0041] In some embodiments, the dosage regimen of the drug will vary based on known factors, such as the pharmacodynamic characteristics of the particular drug and its administration pattern and route; the recipient's age, sex, health, medical condition, and weight; the nature and severity of symptoms; the types of concurrent treatments; the frequency of treatment; the route of administration; and the desired effect. In some embodiments, the drug of the present invention may be administered in a single daily dose, or the total daily dose may be administered in divided doses two, three, or four times daily.

[0042] In some embodiments, administration refers to the method of giving a dose / effective amount of a drug or compound or pharmaceutical composition to a subject (e.g., a patient). Administration can be by any suitable means, including parenteral, intrapulmonary, and intranasal administration, and intralesional administration if intended for local treatment. Parenteral infusion includes, for example, intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration. Depending on whether the administration is transient or long-term, administration can be carried out via any suitable route, such as by injection, such as intravenous or subcutaneous injection. Various dosing schedules are covered in this application, including but not limited to single administration or multiple administrations at multiple time points, bolus administration, and pulsatile infusion.

[0043] In some embodiments, an effective amount or therapeutically effective amount refers to an amount of compound sufficient to provide therapeutic or preventive benefit in the treatment or prevention of a disease, or sufficient to delay or minimize disease-related symptoms. Furthermore, a therapeutically effective amount of a compound used in this application refers to an amount of the compound, alone or in combination with other therapies that provide therapeutic benefit in the treatment or prevention of a disease. When used in conjunction with a compound used in this application, the term may include amounts that improve overall treatment, reduce or avoid symptoms or causes of a disease, enhance the therapeutic efficacy of another therapeutic agent, or synergize with another therapeutic agent. The full therapeutic effect does not necessarily occur with the administration of a single dose (or a single administration), but may occur only after a series of doses. Therefore, an effective amount may be administered in one or more administrations.

[0044] The subject refers to any animal, including both human and non-human animals. Non-human animals include all vertebrates, such as mammals, including non-human primates (especially higher primates), sheep, dogs, rodents (such as mice or rats), guinea pigs, goats, pigs, cats, rabbits, cattle, and any livestock or pets; as well as non-mammals, such as chickens, amphibians, reptiles, etc.

[0045] In some embodiments, treatment refers to reducing or alleviating, improving, or eradicating a disease or one or more symptoms associated with the disease. In some embodiments, the term refers to minimizing the spread or worsening of the disease due to the administration of one or more preventative or therapeutic agents to a patient suffering from the disease. For the purposes of the various aspects and embodiments provided in this application, treatment includes, but is not limited to, reducing, alleviating, or improving one or more clinical manifestations or side effects of the treated disease or condition, improving one or more clinical outcomes, reducing the severity of the disease, delaying or slowing disease progression, improving, alleviating, or stabilizing the disease state, and other beneficial results described in this application.

[0046] The invention is further illustrated below with reference to specific embodiments. It should be understood that the specific embodiments described herein are by way of example and are not intended to limit the invention. The main features of the invention can be used in various embodiments without departing from the scope of the invention.

[0047] Example 1 1. Experimental Materials Cell source: Mouse microglia BV2 and human neuroblastoma cells SH-SY5Y were purchased from Wuhan Pronosei Biotechnology Co., Ltd., and cultured in DMEM high-glucose medium containing 10% FBS, incubated at 37℃ with 5% CO2 saturated humidity, passaged every 2-3 days, and cells in the logarithmic growth phase were used for experiments.

[0048] 2. Experimental Methods Experimental Groups: Control group: treated with an equal volume of 0.01M PBS for 36 hours; MPP + Group: Join MPP + Treatment for 36 hours (0.5 mM, 36 hours); Lumacaftor group: Lumacaftor was added and treated for 36 h (10 μM, 36 h); Lumacaftor+MPP + Group: Add both Lumacaftor and MPP + Treatment for 36 hours (10 μM Lumacaftor, 0.5 mM MPP) + ,36h).

[0049] 3. Experimental Results Lumacaftor can inhibit apoptosis in SH-SY5Y cells. Using Transwell to co-culture BV2 cells and SH-SY5Y cells, it was found that only in MPP cells did this effect occur. + In the single-treatment group, the NF-κB pathway in BV2 cells was upregulated, IL-1R1 in SH-SY5Y cells was significantly upregulated, and apoptosis-related signals cleaved-caspase 3 and the Bax / Bcl-2 ratio were significantly upregulated; however, after simultaneous treatment of BV2 cells with Lumacaftor, the NF-κB pathway declined, and no significant changes were observed in IL-1R1 and apoptosis-related signals in SH-SY5Y cells. Figure 1 ).

[0050] Example 2 1. Experimental Materials Animal source: Healthy male C57BL / 6J mice (8 weeks old, weighing 20-22g) were purchased from Beijing Huafukang Biotechnology Co., Ltd. Animals were housed in the SPF-grade animal facility of the Beijing Institute of Neurosurgery, with a 12-hour light-dark cycle, free access to food and water, and underwent acclimatization for one week before the start of the experiment.

[0051] Drug: Lumacaftor, purchased from Selleck Biotechnology Co., Ltd., USA, catalog number S1565.

[0052] MPTP, purchased from Selleck Biotechnology Co., Ltd., USA, product number S4732.

[0053] 2. Experimental Methods MPTP PD Model Construction Subacute MPTP intraperitoneal injection protocol: Dosage: 20 mg / kg, once daily for 5 consecutive days; Solvent: 0.9% physiological saline, freshly prepared, stored away from light; Control group: treated with an equal volume of physiological saline simultaneously.

[0054] Animal grouping Control group: Intraperitoneal injection of normal saline, followed by intraperitoneal injection of normal saline for 5 consecutive days after 1 week, and behavioral assessment and tissue collection were performed after 1 week; MPTP group: Intraperitoneal injection of normal saline, followed by continuous intraperitoneal injection of MPTP for 5 days after 1 week, and behavioral assessment and tissue collection after 1 week. Lumacaftor group: Lumacaftor 3 mg / kg was injected intraperitoneally, followed by continuous intraperitoneal injection of Lumacaftor for 5 days after 1 week. Behavioral assessment and tissue sampling were performed after 1 week. Lumacaftor+MPTP group: Lumacaftor 3mg / kg was injected intraperitoneally, followed by continuous intraperitoneal injection of Lumacaftor and MPTP for 5 days after 1 week. Behavioral assessment and tissue sampling were performed 1 week later.

[0055] 3. Experimental Results A mouse PD model induced by MPTP was successfully constructed. Immunohistochemical staining revealed a significant decrease in TH-positive nerve fibers in the striatum of the MPTP-treated mice, while treatment with Lumacaftor significantly increased the number of TH-positive nerve fibers. Similarly, a significant decrease in TH-positive neurons in the substantia nigra of the MPTP-treated mice was observed, while treatment with Lumacaftor significantly increased the number of TH-positive neurons. Figure 2 ).

[0056] Example 3 1. Experimental Materials Drug: Lumacaftor, purchased from Selleck Biotechnology Co., Ltd., USA, catalog number S1565.

[0057] MPTP, purchased from Selleck Biotechnology Co., Ltd., USA, product number S4732.

[0058] MPTP PD Model Construction Subacute MPTP intraperitoneal injection protocol: Dosage: 20 mg / kg, once daily for 5 consecutive days; Control group: treated with an equal volume of physiological saline simultaneously.

[0059] Animal grouping Control group: Intraperitoneal injection of normal saline, followed by intraperitoneal injection of normal saline for 5 consecutive days after 1 week, and behavioral assessment and tissue collection were performed after 1 week; MPTP group: Intraperitoneal injection of normal saline, followed by continuous intraperitoneal injection of MPTP for 5 days after 1 week, and behavioral assessment and tissue collection after 1 week. Lumacaftor group: Lumacaftor 3 mg / kg was injected intraperitoneally, followed by continuous intraperitoneal injection of Lumacaftor for 5 days after 1 week. Behavioral assessment and tissue sampling were performed after 1 week. Lumacaftor+MPTP group: Lumacaftor 3mg / kg was injected intraperitoneally, followed by continuous intraperitoneal injection of Lumacaftor and MPTP for 5 days after 1 week. Behavioral assessment and tissue sampling were performed 1 week later.

[0060] 2. Experimental Methods Behavioral analysis Spinning rod experiment: Acceleration mode (5-50 rpm), record the rotational speed during the fall; Pole climbing experiment: Record the turning time (T-turn) and the total climbing time (T-total). Open field experiment: to evaluate parameters such as total distance traveled, time spent at the center, and average speed.

[0061] 3. Experimental Results In the pole climbing experiment, the turning time and total time of mice in the MPTP injection group were significantly prolonged, while no significant motor function impairment was found after Lumacaftor treatment. Figure 3 A, 3B); In the rotundus experiment, it was found that the rotation speed of mice in the MPTP injection group was significantly shortened when they fell from the rotundus, while no significant motor function impairment was found after Lumacaftor treatment. Figure 3C); In open field experiments, the total distance traveled in the open field was significantly reduced, the average speed was significantly reduced, and the number of entries into and dwell time in the central area were significantly reduced. However, no significant motor function impairment was found after Lumacaftor treatment. Figure 3 DH).

[0062] The above description of the embodiments is only for understanding the method and core ideas of the present invention. It should be noted that those skilled in the art can make various improvements and modifications to the present invention without departing from the principles of the invention, and these improvements and modifications will also fall within the protection scope of the claims of the present invention.

Claims

1. Application of Lumacaftor in the preparation of drugs for treating Parkinson's disease.

2. The application according to claim 1, characterized in that, The Lumacaftor treats Parkinson's disease by inhibiting neuronal apoptosis, increasing dopaminergic neurons, and improving motor function through one or more of these mechanisms.

3. The application according to claim 1, characterized in that, The medications also include other medications for treating Parkinson's disease.

4. The application according to claim 3, characterized in that, The drug also includes pharmaceutically acceptable excipients.

5. The application according to claim 4, characterized in that, Pharmaceutically acceptable excipients include diluents, binders, surfactants, humectants, adsorbents, lubricants, fillers, and disintegrants.

6. The application according to claim 5, characterized in that, The dosage forms of the drug include gastrointestinal dosage forms and non-gastrointestinal dosage forms.

7. The application according to claim 6, characterized in that, The gastrointestinal dosage forms include tablets, granules, capsules, solutions, dry suspensions, powders, sustained-release preparations, effervescent tablets, emulsions, suspensions, syrups, drops, and chewable tablets.

8. The application according to claim 6, characterized in that, The non-gastrointestinal drug delivery dosage forms include injection dosage forms, respiratory drug delivery dosage forms, cavity drug delivery dosage forms, mucosal drug delivery dosage forms, and skin drug delivery dosage forms.

9. Application of Lumacaftor in the preparation of drugs that inhibit neuronal apoptosis / increase dopaminergic neurons / improve motor function.

10. A drug for treating Parkinson's disease, characterized in that, The drug includes Lumacaftor; Preferably, the drug also includes other drugs for treating Parkinson's disease; Preferably, the drug further includes pharmaceutically acceptable excipients.

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