WO2019061395A1 - 兴奋性神经毒性相关损伤的治疗肽组合物 - Google Patents
兴奋性神经毒性相关损伤的治疗肽组合物 Download PDFInfo
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- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
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- C07K2319/10—Fusion polypeptide containing a localisation/targetting motif containing a tag for extracellular membrane crossing, e.g. TAT or VP22
Definitions
- the present application relates generally to the medical field, and in particular, the present application provides compositions for treating central nervous system damage and uses thereof.
- Stroke is a common acute cerebrovascular disease in middle-aged and elderly people, and tends to be younger. It is one of the three diseases (cancer, cardiovascular disease, diabetes) that are the most harmful to humans in the world today. China has nearly 3 million deaths from cerebrovascular diseases every year, 4 to 5 times higher than that of European and American countries, 3.5 times that of Japan, and even higher than that of developing countries such as Thailand and India. The incidence rate increases at an annual rate of 8.7%, and the recurrence rate More than 30%, the recurrence rate reached 54% within 5 years; 75% of stroke survivors lost their ability to work and 40% were disabled.
- Stroke can be broadly divided into two categories, namely ischemic stroke and hemorrhagic stroke, of which ischemic stroke accounts for 85% of the total number of stroke patients.
- the therapeutic drugs for ischemic stroke are mainly divided into the following categories: vasodilators (such as dipyridamole), drugs that improve microcirculation, expand blood volume (such as low molecular dextran, etc.), drugs that dissolve thrombus (such as urine).
- Kinase, etc. anticoagulant therapy, drugs that prevent platelet aggregation (such as aspirin), traditional Chinese medicine, neuroprotective agents, etc., but because most of these drugs have side effects, potential risks, or ineffective effects, so research
- drugs that prevent platelet aggregation such as aspirin
- traditional Chinese medicine such as aspirin
- neuroprotective agents such as aspirin
- the pathogenesis of stroke and drug development for its mechanism have important social significance for the prevention and treatment of cerebrovascular disease.
- Stroke is characterized by neuronal cell death in areas of ischemia, cerebral hemorrhage, and/or traumatic areas.
- Neuronal death or injury caused by cerebral ischemia is a process of injury cascade. After cerebral ischemia, blood perfusion decreases, excitatory neurotransmitters increase, and NMDA and AMPA receptors are activated, causing ion channel opening, calcium ion Inflow, activation of a large number of enzymes triggers a signal cascade that causes multiple pathways of neuronal damage.
- Its downstream post-synaptic density 95 protein (PSD-95) triggers a series of ischemic injuries through interaction with various proteins, is a key site for cerebral ischemic injury, and is also a potential target for drug therapy. Therefore, the development of PSD-95 inhibitors has great medicinal significance for neurological damage caused by various excitatory neurotoxicity including stroke.
- NMDA neurodegenerative diseases
- ALS amyotrophic lateral sclerosis
- Huntington's disease various neurodegenerative diseases
- NMDA receptor NMDA receptor
- the onset of epilepsy includes three different and continuous pathophysiological processes, including initiation, maintenance and expansion of episodic discharge, and inhibition of episodes. During this process, excitatory neurotransmitters such as glutamate and aspartate play an important role. .
- PSD-95 In Alzheimer's disease, PSD-95 is involved in the neurotoxic mechanism that leads to it through the GluR6-PSD-95-MLK3 pathway. Furthermore, in Huntington's disease, PSD-95 is a mediator of neurotoxicity of NMDA receptors and huntingtin mutants. Therefore, the development of PSD-95 inhibitors is also important for the treatment, improvement and prevention of the above diseases.
- peptide drugs For peptide drugs, they are limited by the storage conditions of the drug and have limited ability to withstand environmental stress. Peptides may undergo pH changes during high temperature and long-term storage, which may degrade to a certain extent, resulting in decreased purity, dramatic changes in appearance, short storage period, and thus affecting drug efficacy. At the same time, high requirements are imposed on transportation, which limits peptides. The large-scale commercial application of generic drugs requires technical improvements.
- the application provides a pharmaceutical composition comprising a peptide, a pH adjusting agent and a filler, wherein the peptide comprises the amino acid sequence YEKLLDTEI (SEQ ID NO: 1) or a functional variant thereof.
- the functional variant is a variant produced after one or more conservative substitutions of the LDTEI moiety of SEQ ID NO: 1, preferably the conservative substitution is selected between D and E Substitution, substitution between L, V and I and substitution between T and S.
- the functional variant is a variant produced by replacing the LDTEI portion of SEQ ID NO: 1 with any of the following sequences: LDTEL, LDTEV, LDTDI, LDTDL, LDTDV, LDSEI, LDSEL, LDSEV, LDSDI, LDSDL, LDSDV, LETEI, LETEL, LETEV, LETDI, LETDL, LETDV, VDTEI, VDTEL, VDTEV, VDTDI, VDTDL, VDTDV, IDTEI, IDTEL, IDTEV, IDTDI, IDTDL, IDTDV, IETEI, IETEL, IETEV, IETDI, IETDL, IETDVD.
- the peptide is a chimeric peptide comprising the amino acid sequence YEKLLDTEI (SEQ ID NO: 1) or a functional variant thereof and an internalization peptide that facilitates uptake of the chimeric peptide by a cell.
- the internalization peptide comprises the amino acid sequence YGRKKRRQRRR (SEQ ID NO: 2).
- the chimeric peptide comprises the amino acid sequence YGRKKRRQRRR YEKLLDTEI (SEQ ID NO: 3).
- the pH adjusting agent is selected from the group consisting of histidine buffer, arginine buffer, sodium succinate buffer, potassium succinate buffer, sodium citrate buffer, gluconate buffer, acetic acid Salt buffer, phosphate buffer and Tris buffer or any combination of the above, preferably citric acid / disodium hydrogen phosphate buffer or histidine / arginine buffer, more preferably histidine / arginine Buffer.
- the composition has a pH of from about 5.5 to 8, preferably from about 6 to 7.5, more preferably from about 6 to 7, further preferably from about 6.5 to 7, most preferably about 6.5.
- the histidine/arginine content in the histidine/arginine buffer is from about 1% to 10% by weight, preferably from about 3% to 10% by weight.
- the filler is selected from the group consisting of trehalose, mannitol, glucose, lactose, cyclodextrin, dextran-40, sorbitol, sucrose, glycine, or any combination of the above, preferably trehalose, mannitol, glucose, Lactose or any combination of the above is more preferably trehalose.
- the mass ratio of peptide to trehalose is from about 1:0.05 to 1:10, preferably from about 1:0.5 to 1:5, more preferably from about 1:0.8 to 1:3, most preferably about 1:1.
- the filler is trehalose and the pH adjuster is a histidine/arginine buffer.
- the mass ratio of peptide to trehalose is about 1:1.
- the pH of the composition is about 6.5 ⁇ 0.5.
- the composition further comprises a cryoprotectant and/or a surfactant, preferably the cryoprotectant is polyethylene glycol and/or the surfactant is a polysorbate, preferably polysorbate 20 or polysorbate Ester 80.
- the composition further comprises a deamidation inhibitor.
- the pharmaceutical composition is in the form of a pre-lyophilized formulation, or in the form of a lyophilized formulation, or a reconstituted formulation obtained by combining a lyophilized formulation with an aqueous solution.
- the pharmaceutical composition is for treating, ameliorating or preventing a disease in a mammal: a nervous system injury and associated disease or pain caused by the injury, a neurodegenerative disease, anxiety or epilepsy, or as Neuroprotective agent.
- the present application provides a method of treating, ameliorating or preventing a disease in a mammal: a nervous system injury and a disease or pain associated with the injury, a neurodegenerative disease, anxiety or epilepsy, including to an individual in need thereof
- the pharmaceutical composition of the first aspect is administered.
- the present application provides the use of the pharmaceutical composition of the first aspect in the manufacture of a medicament or a neuroprotective agent for treating, ameliorating or preventing a disease in a mammal: a nervous system injury and the injury Related diseases or pain, neurodegenerative diseases, anxiety or epilepsy.
- the disease is a nervous system injury caused by stroke or stroke.
- the stroke comprises an ischemic stroke, a hemorrhagic stroke, and a hemorrhagic stroke converted from an ischemic stroke, preferably the stroke is an ischemic stroke.
- the nervous system injury is a neurological damage caused by excitatory neurotoxicity.
- the injury or pain is located in the peripheral nervous system or the central nervous system.
- the neurological damage caused by excitotoxicity comprises an episode of stroke or spinal cord injury, ischemic or traumatic injury of the brain or spinal cord, and central nervous system (CNS) neurons.
- CNS central nervous system
- Injury including acute CNS injury, ischemic stroke or spinal cord injury, as well as hypoxia, ischemia, mechanical injury and neurodegenerative diseases, anxiety, epilepsy, stroke-induced damage.
- the neurodegenerative disease comprises Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease or Huntington's disease.
- Figure 1 shows the Pull-down assay to detect the interaction of P5 with the PDZ1/2 domain.
- M represents the protein molecular weight marker; Lane 1 is His+PDZ1/2+P5; Lane 2 is P5 alone; Lane 3 is His+P5; Lane 4 is His+PDZ1/2.
- the elution band shown in lane 1 contains both P5 and PDZ1/2, confirming that P5 is capable of binding to the PDZ1/2 domain.
- Figure 2 shows a TTC staining of brain sections of polypeptide P5 versus rat MCAO model.
- Figure 3 is a graph showing the statistical data of cerebral infarction volume in the treatment and prevention of rat MCAO model in different dose groups of polypeptide P5. **p ⁇ 0.01.
- Figure 4 shows the distribution of polypeptide P5 in the rat brain.
- Figure 5 shows rat brain TTC staining.
- Figure 6 shows the results of HE staining of rat brain paraffin sections.
- F YE-NA-1 group
- G P5 group
- H P5 preventive administration group.
- Figure 7 shows the effect of different fillers on the formation and stability of P5 lyophilized formulations (Nos. 0, 1 and 2) on Day 0.
- Figure 8 shows the effect of different fillers on the formation and stability of P5 lyophilized formulations (Nos. 3, 4 and 5) on Day 0.
- Figure 9 shows the effect of different fillers on the formation and stability of P5 lyophilized formulations (Nos. 0, 1 and 2) at week 1.
- Figure 10 shows the effect of different fillers on the formation and stability of P5 lyophilized formulations (Nos. 3, 4 and 5) at week 1.
- Figure 11 shows the results of using the histidine/arginine to adjust the pH of the composition (i.e., pH range screening experiment II) at week 1.
- Figure 12 shows the results of the pH of the composition (i.e., pH range screening experiment II) at week 2 using histidine/arginine.
- compositions including peptides, fillers, pH buffers through extensive research, Such a composition has at least one of the following advantages:
- the product can be kept from degradation during a long storage period, and its appearance, purity and impurity content can meet the requirements of clinical application.
- chimeric peptide denotes a peptide having two component peptides that are not naturally associated with each other, which can be combined as a fusion protein or by chemical bonds.
- PDZ domain refers to a modular protein domain of approximately 90 amino acids characterized by a synaptic protein PSD-95, a Drosophila-separating connexin, a Discs-Large (DLG), and an epithelial tight junction protein Z01. (Z01) has significant (eg, at least 60%) sequence identity.
- the PDZ domain is also known as Discs-Large homology repeats ("DHRs") and GLGF repeats.
- DHRs Discs-Large homology repeats
- the PDZ domain typically displays a retained core consensus sequence (Doyle, D.A., 1996, Cell 85: 1067-76).
- Exemplary PDZ domain-containing proteins and PDZ domain sequences are disclosed in U.S. Patent Application Serial No. 10/714,537.
- NMDA receptor or “NMDAR” refers to a membrane associated protein known to interact with NMDA. These receptors can be human or non-human (eg, mice, rats, rabbits, monkeys, etc.).
- the term "specifically binds” refers to a bond between two molecules (eg, a ligand and a receptor) characterized by one molecule (ligand) and another specific molecule in the presence of many other different molecules ( The ability of a receptor to bind, ie, the ability to display a preferential binding of one molecule to another in a heterogeneous mixture of molecules. Specific binding of the ligand to the receptor is also demonstrated as follows: When an excess of unlabeled ligand is present, the binding of the detectably labeled ligand to the receptor is reduced (i.e., binding competition assay).
- Statistically significant means a p value of ⁇ 0.05, preferably ⁇ 0.01, most preferably ⁇ 0.001.
- a “functional variant” refers to a variant having the same or similar biological function and properties as the parent.
- a “functional variant” can be obtained by performing one or more conservative substitutions in the parent.
- the term "internalized peptide”, also known as a penetrating peptide, is widely used in the field of protein medicine and its function is to promote the uptake and absorption of active peptides bound thereto by cells.
- the internalization peptide can be a Tat peptide, wherein one non-limiting example of a Tat peptide is YGRKKRRQRRR (SEQ ID NO: 2).
- the application provides a pharmaceutical composition
- a pharmaceutical composition comprising a peptide, a pH adjusting agent and a filler, wherein the peptide comprises the amino acid sequence YEKLLDTEI (SEQ ID NO: 1) or a functional variant thereof.
- the peptide is also referred to herein as an "active peptide" which acts as an active moiety in the chimeric peptides of the present application for treating central nervous system damage or as a neuroprotective agent.
- NMDAR-based structures some active peptides that inhibit the interaction between NMDAR and PSD-95 are NMDAR-based structures.
- NMDAR2B has GenBank ID 4099612 and the C-terminal 20 amino acids are FNGSSNGHVYEKLSSLESDV and PL motif ESDV.
- Some of the existing active peptides have selected a partial amino acid sequence at the C-terminus of NMDAR2B, thereby producing competitive inhibition of PSD-95 with NMDAR2B.
- Studies have suggested that the ESDV or LESDV segments in the above peptides play an important role in inhibiting the interaction between NMDAR and PSD-95 proteins.
- a functional variant provided herein is a variant produced following one or more conservative substitutions of the LDTEI moiety of SEQ ID NO: 1.
- the conservative substitution is selected from the group consisting of a substitution between D and E, a substitution between L, V and I, and a substitution between T and S.
- the functional variant is a variant produced by replacing the LDTEI portion of SEQ ID NO: 1 with any of the following sequences: LDTEL, LDTEV, LDTDI, LDTDL, LDTDV, LDSEI, LDSEL , LDSEV, LDSDI, LDSDL, LDSDV, LETEI, LETEL, LETEV, LETDI, LETDL, LETDV, VDTEI, VDTEL, VDTEV, VDTDI, VDTDL, VDTDV, IDTEI, IDTEL, IDTEV, IDTDI, IDTDL, IDTDV, IETEI, IETEL, IETEV , IETDI, IETDL, IETDVD.
- the functional variants disclosed herein further comprise the same as at least 60%, 70%, 75%, 80%, 85%, 90%, 95%, or even higher than the peptides mentioned above.
- Sexual amino acid sequence It is known in the art that "identity" between two proteins is determined by aligning the sequence of a second protein substituted with the amino acid sequence of one protein and its conserved amino acid. The degree of identity between the two proteins is determined using computer algorithms and methods well known to those skilled in the art. The identity between two amino acid sequences is preferably determined by using the BLASTP algorithm.
- the functional variants disclosed herein include substitutions, deletions, additions and/or amino acid residues at 1, 2, 3, 4, 5 or more compared to the peptides mentioned above. Or insert a specific peptide that differs from the above disclosure.
- a functional variant can be distinguished from a particular peptide disclosed above by one or more substitutions, deletions, additions, and/or insertions. These variants may be naturally occurring or may be synthetically produced, for example, by modifying one or more of the above-described peptide sequences disclosed herein and evaluating them according to any of a variety of techniques well known in the art as described herein. Biological activity.
- the peptide is a chimeric peptide comprising the amino acid sequence YEKLLDTEI (SEQ ID NO: 1) or a functional variant thereof and an internalization peptide that facilitates uptake of the chimeric peptide by a cell.
- the internalization peptide suitable for the present application is not limited to a specific species, as long as The purpose of transmembrane and internalization can be achieved. It will also be understood by those skilled in the art that since the target of action of the active peptide is mainly located inside the neuronal cell, it is preferred that the internalization peptide which is specifically adapted to the neuronal cell. In some embodiments, the internalization peptide can be a Tat peptide.
- the amino acid sequence of the Tat peptide is YGRKKRRQRRR (SEQ ID NO: 2).
- the chimeric peptide comprises the amino acid sequence YGRKKRRQRRRYEKLLDTEI (SEQ ID NO: 3).
- the internalization peptide may be linked to the active peptide by an amide bond as a fusion peptide, but may also be joined by other suitable means, such as chemical bond ligation. Coupling of the two components can be achieved by a coupling agent or a conjugating agent.
- a coupling agent or a conjugating agent A large number of such reagents are commercially available and can be found in S. S. Wong, Chemistry of Protein Conjugation and Cross-Linking, CRC Press (1991).
- Some examples of cross-linking reagents include J-succinimide-3-(2-pyridinedithio)propionate (SPOP) or N,N'-(1,3-phenylene)dimale.
- crosslinking reagents include P,P'-difluoro-m,m'-dinitrodiphenyl sulfone (which forms irreversible crosslinks with amino and phenolic groups); dimethyl diethylamine hexanoate (for Amino group is specific); phenol-1,4-disulfonyl chloride (which is mainly reacted with an amino group); 1,6-hexamethylene diisocyanate or diisothiocyanate, or phenylazo-p-diisocyanate (which Mainly reacted with an amino group; glutaraldehyde (which reacts with several different side chains) and double nitrogen benzidine (which reacts mainly with tyrosine and histidine).
- P,P'-difluoro-m,m'-dinitrodiphenyl sulfone which forms irreversible crosslinks with amino and phenolic groups
- dimethyl diethylamine hexanoate for Amin
- the peptides described above can optionally be derivatized (eg, acetylated, phosphorylated, and/or glycosylated) to promote affinity with the inhibitor, promote the ability of the inhibitor to be transported across the cell membrane, or promote stabilization. Sex.
- derivatized eg, acetylated, phosphorylated, and/or glycosylated
- the active peptide of the present application and the fusion peptide fused to the internalization peptide can be synthesized by solid phase synthesis or recombinant methods.
- Peptidomimetics can be synthesized using a variety of protocols and methods described in the scientific literature and patent literature, such as Organic Syntheses Collective Volumes, Gilman et al. (ed.) John Wiley & Sons, Inc., NY, al. - Obeidi (1998) Mol. Biotechnol. 9: 205-223; Hruby (1997) Curr. Opin. Chem. Biol. 1: 14-119; 0 stergaard (1997) Mol. Divers. 3: 17-27; 0 stresh (1996) ) Methods Enzymol. 267: 220-234.
- the pH adjusting agent is selected from the group consisting of histidine buffer, arginine buffer, sodium succinate buffer, potassium succinate buffer, sodium citrate buffer, gluconate buffer, acetic acid Salt buffer, phosphate buffer and Tris buffer or any combination of the above.
- the pH adjusting agent is selected from the group consisting of citric acid/disodium hydrogen phosphate buffer or histidine/arginine buffer.
- the pH adjusting agent is selected from the group consisting of histidine/arginine buffer.
- the pH of the composition is from about 5.5 to 8 (eg, about 5.5, 6, 6.5, 7, 7.5, 8). In some embodiments, the pH of the composition is from about 6 to 7.5. In some embodiments, the pH of the composition is from about 6 to 7. In some embodiments, the pH of the composition is between about 6.5 and 7. In some embodiments, the composition has a pH of about 6.5.
- the histidine/arginine content in the histidine/arginine buffer is from about 1% to 10% by weight (eg, about 1, 2, 3, 4, 5, 6 , 7, 8, 9, 10%). In some embodiments, the histidine/arginine content in the histidine/arginine buffer is between about 3% and 10%.
- the filler is selected from the group consisting of trehalose, mannitol, glucose, lactose, cyclodextrin, dextran-40, sorbitol, sucrose, glycine, or any combination of the above.
- the filler is trehalose, mannitol, glucose, lactose, or any combination of the above.
- the filler is trehalose.
- the mass ratio of peptide to trehalose is from about 1:0.05 to 1:10. In some embodiments, the mass ratio of peptide to trehalose is from about 1:0.5 to 1:5. In some embodiments, the mass ratio of peptide to trehalose is from about 1:0.8 to 1:3. In some embodiments, the mass ratio of peptide to trehalose is about 1:1.
- the filler is trehalose and the pH adjuster is a histidine/arginine buffer.
- the mass ratio of peptide to trehalose is about 1:1.
- the pH of the composition is about 6.5 ⁇ 0.5.
- the composition further comprises a cryoprotectant and/or a surfactant, preferably the cryoprotectant is polyethylene glycol and/or the surfactant is a polysorbate, preferably polysorbate 20 or polysorbate Ester 80.
- the composition further comprises a deamidation inhibitor.
- the pharmaceutical composition is in the form of a pre-lyophilized formulation, or in the form of a lyophilized formulation, or a reconstituted formulation obtained by combining a lyophilized formulation with an aqueous solution.
- the composition can be administered parenterally, intravenously, subcutaneously, intraarterially, intracranically, intrathecally, intraperitoneally, topically, intranasally, or intramuscularly. It is preferably administered intravenously.
- the pharmaceutical composition for parenteral administration is preferably sterile and substantially isotonic.
- a composition comprising an active peptide or a chimeric peptide can be formulated into an aqueous solution, preferably formulated into a physiologically compatible buffer such as Hank's solution, Ringer's solution, or physiological saline or acetate buffer (to reduce injection). Discomfort at the site).
- the solution may contain formulas such as suspending, stabilizing and/or dispersing agents.
- compositions comprising active peptides or chimeric peptides can also be formulated as a depot preparation.
- Such long acting formulations can be administered by implantation (for example subcutaneously or intramuscularly) or by intramuscular injection.
- the compound can be formulated with a suitable polymeric or hydrophobic material (for example, as an emulsion in an acceptable oil) or an ion exchange resin, or as a sparingly soluble derivative, for example, formulated as a slightly soluble solution. Salt.
- the active peptides or chimeric peptides disclosed herein may contain charged side chains or ends, they may be included in any of the above formulations as a free acid or base or as a pharmaceutically acceptable salt.
- Pharmaceutically acceptable salts are those which substantially retain the biological activity of the free base and which are prepared by reaction with a mineral acid. Pharmaceutical salts tend to be more soluble in water and other protic solvents than the corresponding free base forms.
- the active peptide or chimeric peptide is used in an amount effective to achieve the intended purpose (e.g., to reduce the damaging effects of damaging stroke and related conditions).
- a therapeutically effective amount means: treated with an active peptide or chimeric peptide disclosed herein, relative to a central nervous system injury in a control population of a patient (or animal model) not treated with the active peptide or chimeric peptide disclosed herein.
- the amount of active peptide or chimeric peptide sufficient to significantly reduce the damage caused by stroke in the patient (or animal model population).
- Average output (measured by infarct volume or disability index) in a comparable patient control population that was not treated by the methods disclosed herein This amount is also considered therapeutically effective compared to an individual treated patient achieving a better output.
- the amount is also considered to be a therapeutically effective amount if the individual being treated shows 2 or fewer disability in the Rankin scale and 75 or more in the Barthel scale.
- the dose is also considered therapeutically effective if the population of treated patients shows a significant improvement (ie less disability) score distribution on the disability scale compared to comparable untreated populations, see Lees et al. N Engl J Med 2006; 354: 588-600.
- a therapeutically effective regimen represents a combination of a therapeutically effective dose and the frequency of administration required to achieve the above intended purpose. Usually a single application is sufficient.
- a preferred dosage range comprises from 0.001 to 20 [mu]mol of active peptide or chimeric peptide per kg patient body weight within 6 hours after stroke, optionally 0.03 to 3 [mu]mol of active peptide or chimeric peptide per kg patient body weight.
- 0.1-20 [mu]mol of active peptide or chimeric peptide per kg patient body weight is administered within 6 hours.
- 0.1-10 [mu]mol of active peptide or chimeric peptide per kg patient body weight is administered within 6 hours, more preferably about 0.3 [mu]mol of active peptide or chimeric peptide per kg patient body weight administered within 6 hours.
- the dosage range is from 0.005 to 0.5 [mu]mol of active peptide or chimeric peptide per kg patient body weight.
- the dose per kg body weight can be converted from rat to human by dividing by 6.2 to compensate for different surface area: mass ratios.
- Suitable dosages of the active peptide or chimeric peptide for human use may include 0.01 to 100 mg/kg of patient body weight, or more preferably 0.01 to 30 mg/kg of patient body weight or 0.01 to 10 mg/kg, or 0.01 to 1 mg/kg, in grams. .
- the amount of active peptide or chimeric peptide administered depends on the subject being treated, the weight of the subject, the severity of the pain, the mode of administration, and the modulation of the prescribing physician.
- the treatment can be repeated when the symptoms are detectable or even undetectable. Treatment can be provided alone or in combination with other drugs.
- a therapeutically effective dose of an active peptide or chimeric peptide disclosed herein is capable of providing a therapeutic benefit without causing significant toxicity.
- the toxicity of the chimeric peptide can be determined in cell cultures or experimental animals by standard pharmaceutical procedures, for example by measuring LD50 (a dose that kills 50% of the population) or LD100 (a dose that kills 100% of the population). The dose ratio between toxic and therapeutic effects is the therapeutic index. Chimeric peptides or peptidomimetics which exhibit a high therapeutic index are preferred (see, eg, Fingl et al, 1975, In: The Pharmacological Basis of Therapeutics, Chapter 1, page 1).
- the pharmaceutical composition is for treating, ameliorating or preventing a nervous system injury or a disease or pain caused by the injury, or as a neuroprotective agent.
- the nervous system injury is a neurological damage caused by excitotoxicity, wherein the damage is located in the peripheral nervous system or the central nervous system.
- neurological damage caused by excitatory neurotoxicity comprises an episode of stroke or spinal cord injury, ischemic or traumatic injury of the brain or spinal cord, and damage to central nervous system (CNS) neurons, including acute CNS.
- CNS central nervous system
- the pharmaceutical composition is for treating, ameliorating or preventing neurological damage caused by ischemic stroke.
- Stroke is a condition caused by impaired blood flow in the CNS. Possible causes include embolism, bleeding, and thrombosis. Some neuronal cells die immediately due to impaired blood flow. These cells release their component molecules (including glutamate), which then activate the NMDA receptor, which increases intracellular calcium levels and intracellular enzyme levels, resulting in more neuronal cell death ( Excitatory neurotoxicity cascade amplification). The death of the CNS organization is called infarction.
- the infarct volume i.e., the volume of neuronal cells in the brain caused by stroke
- Symptomatic effects depend both on the infarct volume and on where the infarct is located in the brain.
- Rankin Stroke Outcome Scale Rankin, Scott MedJ; 2:200-15 (1957)
- Barthel Index Barthel Index
- the Barthel Index is based on a series of questions about the patient's ability to perform 10 basic activities of daily living, which scores between 0 and 100, with lower scores indicating more disability (Mahoney et al., Maryland State Medical Journal) 14:56-61 (1965).
- stroke severity/output can be measured using the NIH Stroke Scale, available on the World Wide Web at ninds.nih.gov/doctors/NIH_Stroke_Scale_Booklet.pdf.
- the scale is based on the patient's ability to perform 11 sets of functions, including assessing the patient's level of consciousness, movement, feel, and language function.
- Cerebral thrombosis refers to a thrombus (blood clot) that is produced in the obstructed portion of a blood vessel.
- Cerebral embolism usually refers to various emboli in the blood (such as a wall thrombus in the heart, atherosclerotic plaque, fat, tumor cells, fibrocartilage or air, etc.) blocked by blood flow into the cerebral artery.
- Blood vessels when the collateral circulation can not be compensated, cause ischemic necrosis of brain tissue in the blood supply area of the artery, and focal neurological deficit occurs.
- the second important cause of embolism is an irregular heartbeat called arterial fibrillation. It causes a condition in which a blood clot can form in the heart, move and transfer to the brain.
- Other potential causes of ischemic stroke are hemorrhage, thrombosis, arterial or venous severing, cardiac arrest, shock from any cause (including bleeding), and iatrogenic causes, such as cerebral blood vessels or blood vessels leading to the brain. Direct surgical injury or cardiac surgery. Ischemic stroke constitutes approximately 83% of all stroke cases.
- the pharmaceutical composition is for treating, ameliorating or preventing a neurodegenerative disease, anxiety or epilepsy, wherein the neurodegenerative disease comprises Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease or Huntington's disease.
- ALS amyotrophic lateral sclerosis
- Parkinson's disease Huntington's disease.
- the pharmaceutical composition is in the form of a pre-lyophilized formulation, or in the form of a lyophilized formulation, or a reconstituted formulation obtained by combining a lyophilized formulation with an aqueous solution.
- the present application provides a method of treating, ameliorating or preventing a nervous system injury and a disease or pain associated with the injury, a neurodegenerative disease, anxiety or epilepsy, comprising administering to the individual in need thereof the drug of the first aspect combination.
- neurological damage caused by excitatory neurotoxicity comprises an episode of stroke or spinal cord injury, ischemic or traumatic injury of the brain or spinal cord, and damage to central nervous system (CNS) neurons, including acute CNS.
- CNS central nervous system
- the neurodegenerative disease includes Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, or Huntington's disease.
- ALS amyotrophic lateral sclerosis
- Parkinson's disease or Huntington's disease.
- the individual is an individual experiencing an ischemic stroke.
- administering the pharmaceutical composition of the first aspect of the invention is capable of reducing the volume of a portion of the cerebral infarction caused by cerebral ischemia.
- the present application provides the pharmaceutical composition of the first aspect, in the preparation of a medicament for treating, ameliorating or preventing a nervous system injury and a disease or pain associated with the injury, a neurodegenerative disease, anxiety or epilepsy, or Use in neuroprotective agents.
- neurological damage caused by excitatory neurotoxicity comprises an episode of stroke or spinal cord injury, ischemic or traumatic injury of the brain or spinal cord, and damage to central nervous system (CNS) neurons, including acute CNS.
- CNS central nervous system
- the neurodegenerative disease includes Alzheimer's disease, amyotrophic lateral sclerosis (ALS), Parkinson's disease, or Huntington's disease.
- ALS amyotrophic lateral sclerosis
- Parkinson's disease or Huntington's disease.
- the medicament is for treating, ameliorating or preventing neurological damage caused by ischemic stroke.
- the Tat transmembrane peptide YGRKKRRQRRR (SEQ ID NO: 2) was selected and ligated to a different number of amino acids to form a peptide library.
- the chimeric peptide molecules in the peptide library were respectively interacted with the PDZ1/2 domain expressed and purified in vitro, and the polypeptide was initially screened according to the strength of the interaction force.
- the immobilized molecule is PDZ1/2 protein, molecular weight: ⁇ 20kD, concentration: 2mg/ml; molecular phase of mobile phase (analyte): polypeptide to be screened, molecular weight: ⁇ 2kD, concentration: 10mg/ml.
- the CM5 chip was used for fixation using a Biacore 3000 instrument.
- the running buffer was PBS + 0.005% Tween 20. Fixation was carried out using an amino coupling method.
- the concentration of the ligand was 10 ⁇ g/ml.
- the fixing buffer was 10 mM sodium acetate, pH 4.0. Fixed amount: 1400 RU, fixed to flow cells 2.
- the flow rate used was 10 ⁇ l/ml and the ligand was injected for 1 minute.
- 10 mM Gly at pH 2.0 + 2.5 was used as a regenerant, and regeneration was carried out at a flow rate of 30 ⁇ l/min.
- the injection time is 30s.
- Kinetic analysis was performed using the following conditions: control channel: flow cell 1; electrophoresis buffer was PBS; concentration gradient was 6.25n, 12.5n, 25n, 50n, 100n, 200n, 400nM using Kinetic Analysis Wizard mode; injection time It was 1 minute; the dissociation time was 2 min; the flow rate was 30 ⁇ l/min.
- the data was fitted using the Biaevaluation 4.1 software.
- the quasi-sum model is a 1:1 binding model.
- the dissociation constant KD value is inversely proportional to the force.
- control chimeric peptide NA-1 was introduced with the following sequence:
- NA-1 YGRKKRRQRRRKLSSIESDV (SEQ ID NO: 4)
- YE-NA-1 YGRKKRRQRRRYEKLSSIESDV (SEQ ID NO: 5)
- the chimeric peptide P5 was further tested in the following experiments, and in some experiments, NA-1 and YE-NA-1 were used as controls.
- Example 2 Pull-down assay to detect the interaction between P5 and PDZ1/2 domain
- the column was equilibrated with 100 ⁇ l of His beads and 1 ml of MCAC-0 buffer for 5 min. Concussion at 4 °C. The mixture was centrifuged at 5000 g for 1 minute at 4 ° C, and the supernatant was discarded. 1 mg of PDZ1/2 protein was added to the mixture and made up to 1 ml with buffer. The mixture was spun for 1 hour at 4 °C. The mixture was centrifuged at 5000 g for 1 minute at 4 ° C, and the supernatant was discarded. Wash 3 times with 1 ml of MCAC-0 buffer for 5 minutes each time (at 4 ° C, shake wash).
- both the P5 and PDZ1/2 domains were contained in the elution band of the chimeric peptide P5, thereby confirming that the chimeric peptide P5 was able to bind to the PDZ1/2 domain.
- Example 3 Therapeutic effect of chimeric peptide on rat MCAO model
- the preparation of the focal cerebral ischemia-reperfusion model was based on the reversible middle cerebral artery occlusion (MCAO) suture method proposed by Longa and improved according to the anatomical structure of the rat brain to prepare a focal cerebral ischemia-reperfusion model.
- MCAO middle cerebral artery occlusion
- CCA common carotid artery
- ECA external carotid artery
- pterygopalatine 0.26mm monofilament nylon fishing line head 0.5cm with paraffin
- the length of the suture was about 18-20 mm from the CCA bifurcation, depending on the animal's weight, embolization right
- the middle cerebral artery is then sutured, and the end of the suture is fixed to the skin.
- the suture was carefully taken out to form a reperfusion.
- the sham operation was just not inserted into the nylon fishing line, and the rest of the steps were the same as the surgery group.
- the body temperature was maintained at (37 ⁇ 0.5) °C during ischemia and 2 h after reperfusion.
- the success of the model is that the left limb is paralyzed after anesthesia in rats, and the standing is unstable. When turning the tail, turning to one side is the criterion for successful model.
- Neurological deficit signs were scored according to Longa and Bederson's 5-point method and scored 24 hours after waking of the animals. 0 points: no symptoms of nerve damage; 1 point: can not fully extend the contralateral forepaw; 2 points: turn to the opposite side; 3 points: dump to the opposite side; 4 points: can not be self-issued, loss of consciousness. The higher the score, the more serious the animal's behavioral disorder.
- Animals Adult SD rats (Vittalia), SPF grade, body weight 220-250 g, male.
- Instruments and medicines 1 line scissors, 2 eye surgery scissors, 4 bends, 4#, 5# surgical sutures, 6 ⁇ 17 triangular needles, 0.26 mm diameter bolts, and needle holders.
- EnbiP sodium chloride injection (Shijiazhuang Group Enbi Pharmaceutical Co., Ltd.), chloral hydrate, furosemide (20mg / support), gentamicin sulfate (80mg / support), cotton swab, medical tray and so on.
- the peptide was synthesized by Kingsray Biotech.
- the experiment was divided into negative control group, sham operation group, model group, positive drug NBP group, NA-1 group, YE-NA-1 group and P5 group.
- saline, positive drug Enbap, NA-1 group (10 mg/kg), YE-NA-1 (10 mg/kg) group, P5 (10 mg/kg), P5 (3 mg/) Kg), P5 (1 mg/kg) was administered to each group of rats via tail vein injection. No drugs were given to the normal group and the sham control group. Infarct volume calculation
- the rats were killed by decapitation.
- the brain tissue was quickly placed in a refrigerator at -20 °C. After 10 minutes, the brain was placed in a room temperature environment. The brain was placed in a rat brain slice mold, and the olfactory bulb was removed. The cerebellum and the low brain stem were analyzed according to the map. The lesions were cut at a distance of 2 mm and cut into five knives, and cut into six consecutive continuous coronal slices. Then, the brain slices were quickly placed in a 5 ml solution containing 2% TTC, and incubated at 37 ° C for 30 minutes in the dark, and the brain slices were flipped once every 5 minutes.
- each group of brain slices was arranged neatly, photographed and saved, and processed by image analysis system software and counted.
- the infarct area of each brain slice was calculated, multiplied by the thickness of each slice of brain 2 mm, and the infarct area of each brain slice of each animal was multiplied by The thickness is added, which is the volume of cerebral infarction. Volume is expressed as a percentage of the brain's hemisphere to eliminate the effects of cerebral edema.
- the therapeutic administration and prophylactic administration of the sub-high dose (3 mg/kg) of P5 also preferably reduces the volume of cerebral infarction.
- the data showed that the value of reducing infarct volume decreased with increasing dose of P5, and the therapeutic effect was positively correlated with drug concentration.
- the therapeutic effect of the polypeptide YE-NA-1 is significantly better than that of NA-1. According to the inventors' hypothesis, the addition of two amino acids of YE may increase the interaction between the polypeptide and the PDZ1/2 domain, thereby being more than NA-1. Shows better healing results.
- the fluorescent salt-labeled peptide FITC-P5 10 mg/kg was injected into the tail vein, and the rats were sacrificed 12 hours after the administration.
- the brain tissue was quickly removed and placed in a small place.
- fluorescence detection is performed. After the fluorescence detection is completed, the brain tissue is placed in the TTC dye solution for staining to determine the correlation between the ischemic area and the drug distribution. As shown in Figures 4 and 5, the normal mouse brain can be completely stained by TTC with no fluorescence.
- the labeled polypeptide is distributed, and the ischemic site of the ischemic rat brain cannot be stained by TTC, and the fluorescently labeled polypeptide is distributed in the ischemic site of the ischemic region with the middle artery region as the core, suggesting that the polypeptide P5 can be targeted to ischemia.
- the site plays a therapeutic role, and its distribution is positively correlated with the degree of ischemia.
- Example 5 HE staining for observation of histological changes
- the rats in each group were decapitated 24 hours after ischemia, and the coronal sections near the chiasm were sliced to a thickness of about 4 mm.
- the sections were fixed in 10% formalin solution and dehydrated by 70% to 100% alcohol concentration gradient. It was transparent twice in xylene, embedded in paraffin, embedded in wax, fixed on wax paraffin and sliced to a thickness of 4 ⁇ m.
- the wax was completely unfolded and attached to a clean and dry glass slide. Store in a refrigerator at 4 ° C, perform conventional HE staining, and observe the staining results by light microscopy.
- the experimental results are shown in Fig. 6.
- the nucleus of the normal brain tissue is clear, the nucleus is round, and the nuclear membrane is intact.
- the ischemic model group has severe neuronal necrosis, cell swelling and nuclear condensation.
- the cytoplasm was loosely stained and vacuolated; the pathological changes in the P5, 10 mg/kg treatment and prophylaxis group were significantly improved, and the results were better than the positive drugs NBP injection, NA-1 and YE-NA-1, 10 mg/kg.
- the preparation method of the lyophilized preparation is illustrated by using a filler as trehalose and a pH adjuster as an arginine solution, and the remaining lyophilized preparation is prepared in a similar manner.
- Vacuum freeze-drying wherein pre-freezing: -30 ° C for 3 h; sublimation: -20 ° C for 3 h, -10 ° C for 5 h, 5 ° C for 10 h; then drying: 30 ° C for 5 h;
- Example 8 Effect of different fillers on the formation and stability of P5 lyophilized preparations
- the molding conditions and stability of the P5 lyophilized preparation were selected by using different fillers.
- the samples were placed in a stability test chamber at 60 ° C for 2 weeks, and samples were taken for sampling at the 1st and 2nd weekends, respectively.
- trehalose, cyclodextrin, mannitol, lactose and glucose as fillers, the properties of different samples, solution clarity and color, pH value, impurity substances and P5 content were observed.
- test method is visual inspection, and the test object should be white loose lyophilized cake or powder.
- Solution clarity and color Take a sample, add 1ml of water to dissolve it, the solution should be clear and colorless, such as turbidity, compared with No. 1 turbidity standard solution (General Rule 0902 first method), not more concentrated; Color, compared with the yellow No. 1 standard colorimetric liquid (the first method of the Chinese Pharmacopoeia, the fourth general rule of 0901), may not be deeper.
- pH value Take the solution under the clarity and color of the solution, and determine the sample by referring to the Chinese Pharmacopoeia, General Rules of the Wholesale of 0631.
- the measurement was carried out in accordance with high performance liquid chromatography.
- the chromatographic conditions are:
- Elution method gradient elution, 0 to 30 min 5-65% B; flow rate: 1.0 ml/min: column temperature: 36 ° C; detection wavelength: 220 nm, injection amount: 10 ⁇ L.
- trehalose, cyclodextrin, mannitol, and lactose kept the sample in a white loose lyophilized mass at week 1, while the control sample showed slight atrophy and the 5th (glucose) was severe.
- the 5th (glucose) was severe.
- Trehalose, cyclodextrin, and mannitol still kept the sample in a white loose lyophilized mass at week 2, while the control sample showed slight atrophy.
- trehalose was selected as a filler for the next test under the consideration of traits, solution clarity and color, pH value, impurity substances and content.
- Example 9 Effect of the amount of trehalose on the molding and stability of P5 lyophilized preparation
- Table 13 Summary of the results of the trehalose dosage screening experiment II, wherein the filling specification was 5 mg/vial, and a 7 ml vial was used.
- Each sample was placed in a 40 ° C stability test chamber for 3 months, and samples were taken at the end of the first month and the end of the third month, respectively.
- samples were taken at the end of the first month and the end of the third month, respectively.
- trehalose the properties of different samples, solution clarity and color, pH, impurities and content.
- the impurities (maximum single impurity and total impurity) of samples No. 1 to No. 9 at 60 ° C for 1 week or 2 weeks were significantly smaller than the control sample (No. 0), indicating that trehalose has a higher Significant protection.
- the pH of each sample varied from about 6.0 to about 7.0 at 2 weeks at 60 ° C and one month at 40 ° C, indicating the need to add pH to the composition.
- a buffer to stabilize the pH For pH values, the pH of each sample varied from about 6.0 to about 7.0 at 2 weeks at 60 ° C and one month at 40 ° C, indicating the need to add pH to the composition.
- a buffer to stabilize the pH For pH values, the pH of each sample varied from about 6.0 to about 7.0 at 2 weeks at 60 ° C and one month at 40 ° C, indicating the need to add pH to the composition.
- a buffer to stabilize the pH to stabilize the pH.
- Example 10 Effect of different pH on solution clarity and impurity substances in lyophilized preparations
- the pH changes of the different samples are also different.
- the pH value at 2 weeks at 60 ° C did not change significantly; in the pH range, the No. 0 of Experiment I was screened, and the pH value at 2 weeks at 60 ° C did not change significantly.
- the H0 of Experiment II was screened, and the pH value at 2 weeks at 60 °C was significantly changed, and the range of change was 0.5; in the amount of trehalose screening experiment No. 1-5, which was at 2 weeks at 60 °C The pH value changed significantly with a change of 1.0.
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Abstract
Description
嵌合肽 | NA-1 | YE-NA-1 | P5 |
KD(M) | 7.53E-08 | 5.44E-08 | 2.99E-08 |
序号 | 名称 | 规格 | 厂家 | 批号 |
1 | P5 | 注射级 | 杭州中肽合成并制备 | CQ-04-00317 |
2 | 海藻糖 | 药用级 | Pfanstiehl | 38540A |
3 | 羟丙基β环糊精 | 药用级 | 山东滨州智源 | 20160309-1 |
4 | 甘露醇 | 药用级 | 南宁化学制药 | F431C |
5 | 乳糖 | 药用级 | 镇江康富生物 | 20160510 |
6 | 葡萄糖 | 药用级 | 西王药业 | 201605133 |
7 | 乙腈 | 色谱级 | 迈瑞达 | |
8 | 三氟乙酸 | 色谱级 | 迈瑞达 |
实验编号 | 0 | 1 | 2 | 3 | 4 |
P5 | 0.20g | 0.20g | 0.20g | 0.20g | 0.20g |
海藻糖 | / | 50mg | 0.10g | 0.15g | 0.20g |
水 | 2ml | 2ml | 2ml | 2ml | 2ml |
灌装量/瓶 | 100μl | 100μl | 100μl | 100μl | 100μl |
实验编号 | 5 | 6 | 7 | 8 | 9 |
P5 | 0.10g | 0.10g | 0.10g | 0.10g | 0.10g |
海藻糖 | 0.15g | 0.20g | 0.30g | 0.40g | 0.50g |
水 | 2ml | 2ml | 2ml | 3ml | 4ml |
灌装量/瓶 | 100μl | 100μl | 100μl | 150μl | 200μl |
Claims (26)
- 药物组合物,其包含肽、pH调节剂和填充剂,其中所述肽包含氨基酸序列YEKLLDTEI(SEQ ID NO:1)或其功能性变体。
- 如权利要求1所述的药物组合物,其中所述功能性变体为SEQ ID NO:1中的LDTEI部分发生一处或多处保守型取代后产生的变体,优选地,所述保守型取代选自D和E之间的取代,L、V和I之间的取代以及T和S之间的取代。
- 如权利要求2所述的药物组合物,其中所述功能性变体为SEQ ID NO:1中的LDTEI部分被替换为下述任一序列后产生的变体:LDTEL、LDTEV、LDTDI、LDTDL、LDTDV、LDSEI、LDSEL、LDSEV、LDSDI、LDSDL、LDSDV、LETEI、LETEL、LETEV、LETDI、LETDL、LETDV、VDTEI、VDTEL、VDTEV、VDTDI、VDTDL、VDTDV、IDTEI、IDTEL、IDTEV、IDTDI、IDTDL、IDTDV、IETEI、IETEL、IETEV、IETDI、IETDL、IETDV。
- 如权利要求1所述的药物组合物,其中所述肽为包含氨基酸序列YEKLLDTEI(SEQ ID NO:1)或其功能性变体和内化肽的嵌合肽,所述内化肽能促进所述嵌合肽被细胞摄取。
- 如权利要求4所述的药物组合物,其中所述内化肽包含氨基酸序列YGRKKRRQRRR(SEQ ID NO:2)。
- 如权利要求5所述的药物组合物,其中所述嵌合肽包含氨基酸序列YGRKKRRQRRR YEKLLDTEI(SEQ ID NO:3)。
- 如权利要求1-6中任一项所述的药物组合物,其中pH调节剂选自组氨酸缓冲液、精氨酸缓冲液、琥珀酸钠缓冲液、琥珀酸钾缓冲液、柠檬酸钠缓冲液、葡糖酸盐缓冲液、醋酸盐缓冲液、磷酸盐缓冲液、Tris缓冲液或以上的任意组合,优选为柠檬酸/磷酸氢二钠缓冲液或组氨酸/精氨酸缓冲液,更优选为组氨酸/精氨酸缓冲液。
- 如权利要求1-7中任一项所述的药物组合物,其中组合物的pH值为约5.5至8,优选为约6至7.5,更优选为约6至7,进一步优选为约6.5至7,最优选为约6.5。
- 如权利要求7所述的药物组合物,其中组氨酸/精氨酸缓冲液中的组氨酸/精氨酸的含量为以重量计约1%至10%,优选为约3%至10%。
- 如权利要求1-9中任一项所述的药物组合物,其中所述填充剂选自海藻糖、甘露醇、葡萄糖、乳糖、环糊精、右旋糖酐-40、山梨醇、蔗糖、甘氨酸或以上的任意组合,优选为海藻糖、甘露醇、葡萄糖、乳糖或以上的任意组合,更优选为海藻糖。
- 如权利要求10所述的药物组合物,其中所述肽与海藻糖的质量比为约1:0.05至1:10,优选为约1:0.5至1:5,更优选为约1:0.8至1:3,最优选为约1:1。
- 如权利要求1所述的药物组合物,其中填充剂是海藻糖,pH调节剂是组氨酸/精氨酸缓冲液。
- 如权利要求12所述的药物组合物,其中所述肽与海藻糖的质量比为约1:1。
- 如权利要求12所述的药物组合物,其中组合物的pH为约6.5±0.5。
- 如权利要求1-14中任一项所述的药物组合物,还包含冷冻保护剂和/或表面活性剂,优选地,冷冻保护剂为聚乙二醇和/或表面活性剂是聚山梨醇酯,优选聚山梨酯20或聚山梨酯80。
- 如权利要求1-15中任一项所述的药物组合物,还包含脱酰胺作用抑制剂。
- 如权利要求1-16中任一项所述的药物组合物,其中所述药物组合物是预冻干制剂的形式,或冻干制剂的形式,或冻干制剂与水溶液相结合而获得的复原制剂的形式。
- 如权利要求1-17中任一项所述的药物组合物,其用于治疗、改善或预防哺乳动物中的下述疾病:神经系统损伤及该损伤引起的相关的疾病或疼痛、神经退行性疾病、焦虑或癫痫,或者用作神经元保护剂。
- 治疗、改善或预防哺乳动物中的下述疾病的方法:神经系统损伤及该损伤相关的疾病或疼痛、神经退行性疾病、焦虑或癫痫,包括向有需要的个体给予权利要求1-17中任一项所述的药物组合物。
- 权利要求1-17中任一项所述的药物组合物在制备用于治疗、改善或预防哺乳动物中的下述疾病的药物或神经元保护剂中的用途:神经系统损伤及该损伤相关的疾病或疼痛、神经退行性疾病、焦虑或癫痫。
- 如权利要求18所述的药物组合物或者权利要求19所述的方法或者权利要求20所述的用途,其中所述疾病为脑卒中或脑卒中导致的神经系统损伤。
- 如权利要求21所述的药物组合物、方法或用途,其中所述脑卒中包括缺血性卒中、出血性卒中和由缺血性卒中转化成的出血性卒中,优选地,所述脑卒中为缺血性卒中。
- 如权利要求18所述的药物组合物或者权利要求19所述的方法或者权利要求20所述的用途,其中所述神经系统损伤为兴奋性神经毒性引起的神经系统损伤。
- 如权利要求18所述的药物组合物或者权利要求19所述的方法或者权利要求20所述的用途,其中所述损伤或疼痛位于外周神经系统或中枢神经系统。
- 如权利要求18所述的药物组合物或者权利要求19所述的方法或者权利要求20所述的用途,其中所述兴奋性神经毒性引起的神经系统损伤包括选自脑卒中或脊髓损伤、脑或脊髓的缺血性或创伤性损伤以及中枢神经系统(CNS)神经元的损伤,包括急性CNS损伤、缺血性脑卒中或脊髓损伤,以及缺氧、缺血、机械损伤和神经退行性疾病、焦虑、癫痫、脑卒中引起的损伤。
- 如权利要求18所述的药物组合物或者权利要求19所述的方法或者权利要求20所述的用途,其中所述神经退行性疾病包括阿尔茨海默氏病、肌萎缩性侧索硬化症(ALS)、帕金森氏病或亨廷顿氏病。
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KR1020207012730A KR102545825B1 (ko) | 2017-09-30 | 2017-09-30 | 흥분성 신경 독성 관련 손상 치료를 위한 펩타이드 조성물 |
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BR112020006321-5A BR112020006321A2 (pt) | 2017-09-30 | 2017-09-30 | composição de peptídeos para tratamento de lesões relacionadas a neurotoxicidade excitatória |
US16/652,323 US11541098B2 (en) | 2017-09-30 | 2017-09-30 | Peptide composition for treating excitatory neurotoxicity related injuries |
PCT/CN2017/104751 WO2019061395A1 (zh) | 2017-09-30 | 2017-09-30 | 兴奋性神经毒性相关损伤的治疗肽组合物 |
EA202090802A EA202090802A1 (ru) | 2017-09-30 | 2017-09-30 | Пептидная композиция для лечения повреждений, связанных с возбуждающей нейротоксичностью |
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WO2024131844A1 (zh) * | 2022-12-23 | 2024-06-27 | 拜西欧斯(北京)生物技术有限公司 | 包含莰醇和肽的缀合物及其用途 |
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CN111132687A (zh) | 2020-05-08 |
AU2017433643A1 (en) | 2020-05-14 |
KR102545825B1 (ko) | 2023-06-22 |
JP2020536065A (ja) | 2020-12-10 |
CN111132687B (zh) | 2023-05-02 |
AU2017433643B2 (en) | 2022-04-28 |
KR20200066334A (ko) | 2020-06-09 |
EP3693001A1 (en) | 2020-08-12 |
BR112020006321A2 (pt) | 2020-11-17 |
EP3693001A4 (en) | 2021-07-07 |
US11541098B2 (en) | 2023-01-03 |
US20210000909A1 (en) | 2021-01-07 |
EA202090802A1 (ru) | 2020-08-05 |
ZA202002329B (en) | 2022-12-21 |
JP7073486B2 (ja) | 2022-05-23 |
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