CN111956799A

CN111956799A - Application of anti-PD-1 antibody and/or PD-L1 antibody in preparation of medicine for treating Parkinson's disease

Info

Publication number: CN111956799A
Application number: CN202010984636.XA
Authority: CN
Inventors: 任宇皓; 蔡慧芝
Original assignee: Biological Antibiotic Co
Current assignee: Biological Antibiotic Co
Priority date: 2020-09-18
Filing date: 2020-09-18
Publication date: 2020-11-20

Abstract

The invention relates to the technical field of medicines, in particular to application of an anti-PD-1 antibody and/or an anti-PD-L1 antibody in preparation of a medicine for treating Parkinson's disease. The invention discovers that the anti-PD-1 antibody and/or the anti-PD-L1 antibody can reduce alpha synuclein aggregation, and the anti-PD-1/PD-L1 antibodies pembrolizumab, nivolumab, cimeprinizumab, terieprinizumab, certralizumab, carprilizumab, tirlizumab, attelizumab, Devolumab and Arvinlizumab can reduce alpha synuclein aggregation. In a specific clinical application case, the anti-PD-1/PD-L1 antibody is found to improve the Parkinson's disease and effectively prevent the development of the disease condition through intravenous injection or oral administration.

Description

Application of anti-PD-1 antibody and/or PD-L1 antibody in preparation of medicine for treating Parkinson's disease

Technical Field

The invention relates to the technical field of medicines, in particular to application of an anti-PD-1 antibody and/or an anti-PD-L1 antibody in preparation of a medicine for treating Parkinson's disease.

Background

Parkinson's disease is a neurodegenerative disease. It becomes the second most common neurodegenerative disease after senile dementia. Over ten million people worldwide are suffering from this Disease (Ball N, Teo W, Chandra S and Chapman j. Parkinson' S Disease and the environment. front nerve 2019; 10: 218.). Its main symptoms are tremor, bradykinesia, muscular tension, heaviness, weakness, impaired posture and balance, constipation, facemasks, stooping, bowing back and changing of speaking and writing postures. The cause of the disease is still unclear. It is generally believed that in the midbrain, abnormal aggregation of alpha Synuclein leads to the failure of dopamine-producing cells to function properly, and even death, leading to a decrease in midbrain dopamine, disorganized motor control, and parkinsonism (Stefanis L. alpha. -Synuclein Parkinson's disease. Cold Spring Harb Perspectrum Med. 2012;2: a 009399.).

Effective treatment of parkinson's disease is by increasing the dopamine concentration in the brain striatum or by mimicking the dopamine effect using drugs such as levodopa, dopamine agonists, MAO-B inhibitors, COMT inhibitors and amantadine. However, there is currently no method to Slow down and Stop the progression of Parkinson's Disease (Foltynie T, Langston JW. Therapies to Slow, Stop, or Reverse Parkinson's Disease.J Parkinsons Dis2018, 8(S1): S115-S121). The development of new drugs to slow, stop and cure the disease has become an urgent issue to be solved.

PD-1 (programmed death receptor 1), also known as CD279 (cluster of differentiation 279), is a protein found on the surface of T cells that functions as an immune checkpoint to down regulate the immune response. anti-PD-1 monoclonal antibodies, such as pembrolizumab, nivolumab, cimiraprimab (cemipimab), terlipiraprimab (toreplimab), sintilimab (sintilimab), carpriclizumab (camrelizumab), and tirilezumab (tiselizumab) have been approved for cancer treatment by FDA and NMPA. PD-L1 (programmed death ligand 1), also known as CD274 (cluster of differentiation 274), is a ligand for PD-1 and is an immunologically downregulated transmembrane protein. It is found in activated T cells, B cells, bone marrow cells and inflammatory macrophages. anti-PD-L1 antibodies, such as Antilizumab (atezolizumab), Dewar mab (durvalumab) and Arvinizumab (avelumab) have been approved by FDA and NMPA for cancer therapy (Qin W, Hu L, Zhang X, et al. The diversion Function of PD-1/PD-L Path facility and cancer. Front immunological. 2019; 10: 2298; Pan C, Liu H, Robins E, Song W, Liu D, Li Z, Zheng L. next-generation immunological agents: recovery surgery in cancer immunological therapy J Hematol. 13: 29).

anti-PD-1/PD-L1 antibodies have been used in cancer therapy, but have not been used as a precedent for the treatment of Parkinson's disease.

Disclosure of Invention

The purpose of the present invention is to provide novel medical uses of an anti-PD-1 antibody and an anti-PD-L1 antibody.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides an application of an anti-PD-1 antibody and/or an anti-PD-L1 antibody in preparing a medicament for treating Parkinson's disease.

In a further embodiment, the medicament comprises an effective dose of a pharmaceutically effective ingredient and a pharmaceutically acceptable carrier; the effective component is an anti-PD-1 antibody and/or an anti-PD-L1 antibody, or the effective component is formed by combining an anti-PD-1 antibody and/or an anti-PD-L1 antibody and other medicaments for treating the Parkinson's disease.

In a further embodiment, the medicament comprises a pharmaceutically effective dose of an anti-PD-1 antibody and/or an anti-PD-L1 antibody and a pharmaceutically acceptable carrier.

In a further aspect, the anti-PD-1 antibody comprises: pembrolizumab, nivolumab, cimepril mab, terlipril mab, sillizumab, carprillizumab, and tirilelizumab; the anti-PD-L1 antibody comprises: amilizumab, Devolumab, and Arvelizumab.

In a further embodiment, the drug is administered by intravenous injection or orally.

In a further embodiment, the effective dose of the anti-PD-1 antibody is administered by intravenous injection of the drug: pembrolizumab for 40-1000 mg/3 weeks or 80-2000 mg/6 weeks; nivolumab, 48-1200 mg/2 weeks or 96-2400 mg/4 weeks; cimetipril monoclonal antibody, 70-1750 mg/3 weeks; terepril monoclonal antibody, 0.6-15 mg/kg/2 weeks; 40-1000 mg/3 weeks of Xindilizumab; 40-1000 mg/2 weeks for Carayleigh monoclonal antibody and 40-1000 mg/3 weeks for tirezumab. When the anti-PD-1 antibody is orally taken, the effective dose of the anti-PD-1 antibody is 3 times of the intravenous injection dose of the corresponding antibody.

In a further embodiment, the effective dose of the anti-PD-L1 antibody is administered by intravenous injection of the drug: 168-4200 mg/2 weeks or 240-6000 mg/3 weeks or 336-8400 mg/4 weeks of the amitrazumab, 2-50 mg/kg/2 weeks of the Dewaruzumab and 160-4000 mg/2 weeks of the aralizumab. When the anti-PD-L1 antibody is orally taken, the effective dose of the anti-PD-L1 antibody is 3 times of the intravenous dose of the corresponding antibody.

In a further embodiment, the drug may be formulated into an emulsion, microemulsion, nanoemulsion, nanoparticle, liposome, lipid nanoparticle, microsphere, microcapsule, nanocapsule, capsule, or the like for oral administration to reduce gastrointestinal damage and increase absorption.

The invention has the beneficial effects that:

the invention discovers that the anti-PD-1 antibody and/or the anti-PD-L1 antibody can reduce alpha synuclein aggregation, and further confirms that the anti-PD-1/PD-L1 antibody is found in the steps of: pembrolizumab, tiramizumab, nivolumab, cimeprinizumab, caprolizumab, tereprinizumab, sillizumab, and also attelizumab, alevrilizumab, Dewaruzumab all reduce alpha synuclein aggregation.

In a specific clinical application case, the anti-PD-1/PD-L1 antibody can improve the Parkinson's disease and can effectively prevent the development of the disease.

Detailed Description

The technical solution of the present invention will be described in detail with reference to the following specific examples.

The experimental animals and experimental drugs used in the examples of the present invention are commercially available.

EXAMPLE 1 Effect of anti-PD-1/PD-L1 antibodies on alpha-synuclein oligomers in Parkinson's mice

In this example, Parkinson's mice were induced using the method of Luk et al (Luk KC, Kehm V, Carroll J, Zhang B, O' Brien P, Trojanowski JQ, Lee V. Pathological α -Synuclein Transmission in Nontransgenic Mice. 2012; 338: 949-. The process is as follows: mu.g of alpha-synuclein fibrils were injected into male C57BL/6 murine striatum. After 60 days, the male C57BL/6 mice were divided into groups of 10 mice each, and the drugs to be tested were injected intravenously: pembrolizumab, tiramizumab, nivolumab, cimeprimab, carprilizumab, tereprimab, certralimab and attelizumab, alvinimab, de waruzumab were injected at two doses for each drug, the specific injection dose is shown in table 1, and the injections were performed once every 10 days, and male C57BL/6 mice injected with PBS were additionally used as a blank control group. On day 95 after injection of α -synuclein fibrils, cerebrospinal fluid from mice was collected and α -synuclein oligomers were detected.

Alpha-Polynucleoprotein oligomer content determination according to the method of EI-Agnaf et al (El-Agnaf OM, Salem SA, Paleologou KE, Curran MD, Gibson MJ, Coirt JA, Schlossman MG, Allop D. Detection of Oligomeric formations of Alpha-Synuclein Protein in Human Plasma a positional Biomarker for Parkinson's disease. FASEB J. 2006; 20: 419-25.). The method is ELISA method, specifically, mAb 211 antibody is dissolved in 200 mM NaHCO₃0.02% (w/v) sodium azide to 1 μ g/mL in a pH 9.6 solution, and then the solution was added to an ELISA plate at 100 μ L/well overnight at 4 ℃. Four washes with PBST wash (PBS containing 0.05% Tween 20). After that, 200 μ L/well of blocking buffer (the blocking buffer comprises 2.5% gelatin and 0.05% Tween 20) was added, and incubated at 37 ℃ for 2 hours. And washed 4 times with PBST wash. 100 μ L cerebrospinal fluid samples were then added to each well and bathed for 2 hours at 37 ℃. The cells were washed again with PBST wash solution 4 times, and 1 mug/mL biotin-labeled mAb 211 diluted in blocking buffer was added to 100 μ L/well and incubated for 2 hours at 37 ℃. Washed four times with PBST wash. 100 muL/well of avidin-coupled alkaline phosphatase diluted in blocking buffer at a ratio of 3:5000 was added, and incubated at 37 ℃ for 1 hour. And washed 4 times with PBST wash. Adding 100 muL/well enzyme substrate pNPP, and acting for 30 minutes at room temperature. Finally, the absorbance was measured at 405 nm.

Reference is made to Lim for the Collection of spinal Fluid in the rat brain (Lim NK, Moestrup V, Zhang X, Wang WA, M Broglier A, Huang FD. An Improved Method for the Collection of Cerebrospinal Fluid from enriched Mice. J Vis Exp. 2018; 133: 56774.).

The clearance of alpha-synuclein oligomers was calculated according to the following equation:

alpha-synuclein oligomer clearance (%) = (absorbance of placebo-absorbance of test drug set)/absorbance of placebo 100%

TABLE 1 Effect of anti-PD-1 antibodies on alpha-synuclein oligomer scavenging in Parkinson's mice

Group of	Dosage (mg/kg)	Alpha-synuclein oligomer clearance (%)
			Pembrolizumab	1	19 ± 4
Pembrolizumab	4	52 ± 6
			Tirezol monoclonal antibody	1	18 ± 2
Tirezol monoclonal antibody	4	57 ± 5
			Nivolumab	1.8	18 ± 2
Nivolumab	7.2	59 ± 6
			Cemifepril monoclonal antibody	1.4	22 ± 3
Cemifepril monoclonal antibody	6.8	58 ± 5
			Karayleigh bead monoclonal antibody	1.5	23 ± 3
Karayleigh bead monoclonal antibody	6	57 ± 7
			Terepril monoclonal antibody	1.1	20 ± 3
Terepril monoclonal antibody	4.5	57 ± 6
			Xindilizumab	1	18 ± 2
Xindilizumab	4	59 ± 7

n =3 (representing the number of repetitions as 3 times)

As can be seen from Table 1, all antibodies had significant effect of eliminating alpha-synuclein oligomers at high concentrations, with clearance of 50% or more. The effect of eliminating alpha-synuclein oligomer is not obvious (p is more than or equal to 0.05) among antibodies at high concentration. The effect of eliminating alpha-polynuclear protein oligomer of all antibodies at low concentration is lower than that at high concentration, and the clearance rate is less than or equal to 25 percent. The effect of eliminating alpha-synuclein oligomer is not obvious (p is more than or equal to 0.05) at low concentration.

TABLE 2 Effect of anti-PD-L1 antibody on alpha-synuclein oligomer scavenging in Parkinson's mice

Group of	Dosage (mg/kg)	Alpha-synuclein oligomer clearance (%)
			Alitilide beads	6	23 ± 4
Alitilide beads	24	53 ± 6
			Arviny	6	20 ± 3
Arviny	24	51 ± 5
			Dewaru	2.5	19 ± 2
Dewaru	10	59 ± 5

n =3 (representing the number of repetitions as 3 times)

As can be seen in Table 2, all antibodies had significant effect of eliminating alpha-synuclein oligomers at high concentrations, with clearance rates of greater than or equal to 50%. The effect of eliminating alpha-synuclein oligomer is not obvious (p is more than or equal to 0.05) among antibodies at high concentration. The effect of eliminating alpha-polynuclear protein oligomer of all antibodies at low concentration is lower than that at high concentration, and the clearance rate is less than or equal to 25 percent. The effect of eliminating alpha-synuclein oligomer is not obvious (p is more than or equal to 0.05) at low concentration.

In general, the anti-PD-1 antibodies pembrolizumab, tiragluzumab, nivolumab, cimeprituril mab, carprilizumab, tereprituril mab, and the anti-PD-L1 antibodies, attelizumab, alvinizumab, and delauzumab all cleared the alpha-synuclein oligomers. This is probably because the anti-PD-1/PD-L1 antibody inhibits PD-1/PD-L1, increases the immune activity of the body, thus blocking the aggregation of alpha synuclein and even destroying the alpha synuclein aggregates, thereby restoring the normal function of dopamine cells and reducing their death, stopping further reduction of dopamine in the brain, and showing the effect of treating parkinson's disease.

EXAMPLE 2 preparation of oral emulsions

The preparation method of the emulsion comprises the following steps: anti-PD-1 antibody and/or anti-PD-L1 antibody were raised at a rate of 50 mg: 1 mL of the aqueous phase was dissolved in 4% Tween-80 in PBS buffer as follows: dissolving 500mg of pembrolizumab in 10mL of PBS buffer solution containing 4% Tween-80 to serve as a water phase; using olive oil containing 6% lecithin as oil phase; then the water phase and the oil phase are mixed in equal volume, and stirred for 5 minutes at 30000 revolutions after 2 minutes of oscillation at 3000 revolutions, and the mixture is ready for use.

EXAMPLE 3 clinical trials

The Parkinson Disease comprehensive assessment Scale (MDS-UPDRS) was used for clinical assessment (reference https:// www.movementdisorders.org/MDS/MDS-Rating-Scales/MDS-united-Parkinsons-Disease-Rating-Scale-MDS-MDS-UPDRS. htm). The experiment lasts for 12 months. Patients with Parkinson's disease are diagnosed by a professional doctor as an inclusion standard. Patients with levodopa-drug tolerance were excluded from this experiment. In the embodiment, the dosage of the levodopa medicament used as the Parkinson disease treatment medicament in the early stage is properly reduced before the experiment so that the symptoms are obvious. The anti-PD-1/PD-L1 antibody drug used in this example was a commercial clinical drug, and the use and amount by intravenous injection were referred to the use amount thereof in anticancer therapy.

Case 1:

female, age: age 78, body weight: 65 kg, height: 1.60 m, six years ago, was diagnosed as Parkinson's disease. The main symptoms are: slight tremor of the limbs, head and neck pain, soreness of the waist and back pain, facial mask, weakness, bradykinesia, muscular tension, watery mouth, constipation, stooping, bowing back and occasional restlessness. The patient normally takes 500mg of the baysizine tablet and 50mg of the piribedil every day, and the main symptoms are mild by matching with certain antidepressant. Three days before the experiment, 250mg of the baysizine tablets are taken every day, and the main symptoms before the experiment is taken appear if other medicines are used as usual.

The experimental procedure was as follows:

an anti-PD-1 antibody, pembrolizumab, 200mg was injected intravenously. After 24h, the main symptoms are obviously improved, and compared with before and after treatment, MDS-UPDRS is improved by 54%. And the effect is maintained for about three weeks. Symptoms reappear after three weeks. The injection of pembrolizumab 200mg once more, after 24h, the primary symptoms improved significantly, and the effect remained for about three weeks.

After the symptom reappears for three days, the pembrolizumab 600mg is orally taken, and after 24 hours, the main symptom is obviously improved, and compared with before and after treatment, MDS-UPDRS is improved by 56%. And the effect can be maintained for about three weeks. The symptoms reappear. I.e., 600mg pembrolizumab was orally administered, followed by 600mg once every three weeks. The antibody is stopped after three months, and the administration effect can be kept for five weeks.

Case 2:

male, age: age 69, body weight: 66 kg, height: 1.72 m, hypertension more than ten years. Approximately eight years ago, it was diagnosed as parkinson's disease. The main symptoms are: tremor on the left, head and neck pain, bradykinesia, muscular tension, mask face, constipation, stooping, back and bow and depression. The patient takes 625mg of the bazedoary hydrazine in combination with certain antidepressants and antihypertensives every day. Three days before the experiment, 375mg of the polyhydrazine is taken by the patient every day, and other medicines are taken as usual.

The experimental procedure was as follows:

the anti-PD-L1 antibody was injected intravenously at 10mg/kg Devolumab. After 24h, the chief complaints improved significantly and the effect remained for about two weeks. Compared with the treatment before and after, the MDS-UPDRS is improved by 60 percent. Symptoms reappear after two weeks. After another intravenous injection of Devolumab 10mg/kg, the main symptoms are obviously improved after 24h, and the effect can be kept for about 2 weeks.

After two weeks the symptoms reappear three days later, Devolumab 30mg/kg is orally administered. After 24h, the main symptoms are obviously improved, and compared with before and after treatment, the MDS-UPDRS is improved by 58 percent. And the effect can be maintained for about two weeks. After the symptoms reappear, the Devolumab is orally taken at 30mg/kg, and then the Devolumab is orally taken once every two weeks at each time of 30 mg/kg. The antibody is stopped after three months, and the administration effect can be kept for four weeks.

Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention. The present invention is capable of other embodiments, and various changes and modifications may be made by one skilled in the art without departing from the spirit and scope of the invention.

Claims

1. Use of an anti-PD-1 antibody and/or an anti-PD-L1 antibody for the manufacture of a medicament for the treatment of parkinson's disease.

2. The use of claim 1, wherein the medicament comprises an effective amount of a pharmaceutically active ingredient and a pharmaceutically acceptable carrier; the effective component is an anti-PD-1 antibody and/or an anti-PD-L1 antibody, or the effective component is formed by combining an anti-PD-1 antibody and/or an anti-PD-L1 antibody and other medicaments for treating the Parkinson's disease.

3. The use of claim 1, wherein the anti-PD-1 antibody comprises: pembrolizumab, nivolumab, cimepril mab, terlipril mab, sillizumab, carprillizumab, and tirilelizumab; the anti-PD-L1 antibody comprises: amilizumab, Devolumab, and Arvelizumab.

4. The use according to claim 3, wherein the medicament is administered intravenously or orally.

5. The use of claim 4, wherein the effective dose of the anti-PD-1 antibody is determined by intravenous injection of the drug: pembrolizumab for 40-1000 mg/3 weeks or 80-2000 mg/6 weeks; nivolumab, 48-1200 mg/2 weeks or 96-2400 mg/4 weeks; cimetipril monoclonal antibody, 70-1750 mg/3 weeks; terepril monoclonal antibody, 0.6-15 mg/kg/2 weeks; 40-1000 mg/3 weeks of Xindilizumab; 40-1000 mg/2 weeks for the Cayleigh monoclonal antibody and 40-1000 mg/3 weeks for the tirezumab;

effective dose of anti-PD-L1 antibody when the drug is administered intravenously: replacing the cetirizumab for 168-4200 mg/2 weeks, 240-6000 mg/3 weeks or 336-8400 mg/4 weeks; devolumab 2-50 mg/kg/2 weeks and Arvinizumab 160-4000 mg/2 weeks;

when the medicine is taken orally, the effective dose of the anti-PD-1 antibody or the anti-PD-L1 antibody is 3 times of the intravenous dose of the corresponding antibody.