Belysin is Cima Science’s branded trademark of a proprietary blend of Nicotinamide mononucleotide(NMN) and L-Alpha glycerylphosphorylcholine (Alpha GPC). The invention provides a composition comprising β-Nicotinamide mononucleotide (NMN) and l-α-Glycerolcholinephosphate(α-GPC), wherein the preferred dosage for β- nicotinamide mononucleotide is 3 ~ 9mg/kg/D, while for L-α-GPC is 8 ~ 20mg/kg/d. In the classic Alzheimer’s disease efficacy evaluation model – Morris water maze test model, the composition of the invention can significantly improve the learning and memory behavior of rats after artificial modeling. The compositions of the invention are food-grade raw materials, safe and reliable, and can be taken for a long time.

The bullet key points for the patent are summarized as below:

  1. A composition for treating Alzheimer’s disease, which is characterized in that it comprises β-Nicotinamide mononucleotides NMN and l-α-Glycerol choline phosphate α-GPC, where, the dosage of β-nicotinamide mononucleotide is 3 ~ 9mg /kg/day, The dosage of L-α-glycerol choline phosphate is 8 ~ 20mg /kg/day.
  2. A pharmaceutical preparation for treating Alzheimer’s disease, which is characterized in that it comprises the composition for treating Alzheimer’s disease described in claim 1 and pharmaceutically acceptable excipients.
  3. The pharmaceutical preparation according to claim 2, is a tablet or capsule.
  4. A functional food comprising the composition for treating Alzheimer’s disease according to claim 1 and an edible excipient.
  5. Application of the composition of claim 1 in preparing a drug for treating Alzheimer’s disease.
  6. The composition of claim 1, wherein toxic A is injected into the improved brain β Application of polypeptide induced cognitive behavior in Alzheimer’s rats.

Technical field

The invention relates to a composition for treating Alzheimer’s disease and its application.

Background technology

Alzheimer’s disease (hereinafter referred to as AD) was first discovered and named by German psychiatrist and neuropathologist Alzheimer Alois in 1906. It is a chronic neurodegenerative disease. The main clinical manifestations of Alzheimer’s disease are gradually decreased memory, cognitive impairment, abnormal behavior, and social disorder. With the increase of the average life span of human beings, the incidence rate is increasing year by year. Alzheimer’s disease is about 70% of senile dementia, and there is mixed dementia with vascular dementia. Its main pathological features are brain atrophy, senile plaques in brain tissue, cerebrovascular deposits, and neurofibrillary tangles.

Current studies have shown that the main pathological features of Alzheimer’s disease patients are β-amyloid (Aβ) aggregation into senile plaques, abnormal intracellular Tau protein aggregation forming neuronal fiber tangles (NFT), and neuronal death. In recent years, for the pathogenesis of Alzheimer’s disease, based on the early cholinergic neuron hypothesis, Aβ toxicity hypothesis, and Tau protein hypothesis, the relatively less studied inflammation hypothesis, insulin hypothesis, oxidative imbalance hypothesis, and gene mutation hypothesis have also received increasing attention. The specific pathogenesis of Alzheimer’s disease has not been fully studied yet, except for the existence of several hypotheses, including the cholinergic neuron hypothesis, the Aβ toxicity hypothesis, the Tau protein hypothesis, the insulin hypothesis, and the free radical damage hypothesis. However, Alzheimer’s disease is a complex disease triggered by a combination of genetic and environmental factors, and a single hypothesis cannot explain all the pathogenetic features of Alzheimer’s disease. Since the causative factors of Alzheimer’s disease are very complex. All the hypotheses reviewed in the paper may not fully generalize them, and some mechanisms that are still in the preliminary stage of research or have not been discovered, such as the mitochondrial dynamics imbalance hypothesis, Ca concentration imbalance hypothesis, and microglia influence hypothesis, may be able to enrich the description of the pathogenic factors of Alzheimer’s disease. Until the pathogenesis of Alzheimer’s disease is fully understood, the treatment of Alzheimer’s disease can only be symptomatic, which does not cure the root cause of Alzheimer’s disease. Therefore, finding drugs that can reverse the disease process is the key to overcome Alzheimer’s disease. The lessons learned from numerous failed clinical trials suggest that single-target therapies are not effective for a complex disease like Alzheimer’s.

The cause of Alzheimer’s disease has not yet been elucidated, and no specific drugs have been developed to treat the disease, making the treatment of Alzheimer’s disease a very difficult problem. The development of related drugs has been the pursuit of the industry. In addition, since the incidence of Alzheimer’s disease is increasing every year, preventive measures for the disease are also very important. The development of functional foods with Alzheimer’s therapeutic effects that can be taken safely over a long period of time also holds great promise for the market.

Content of the Invention

The purpose of the present invention is to develop a composition for the treatment of Alzheimer’s disease with a definite effect and convincing safety to meet the need for a composition that can be suitable for long-term administration and can prevent or improve Alzheimer’s disease. The inventors of the present invention completed the present invention after intensive research and unexpectedly discovered the therapeutic effect of the symptoms of Alzheimer’s disease shown by combining NMN and α-GPC. The compositions of the present invention can be applied in the preparation of drugs for the treatment of Alzheimer’s disease, or functional foods for the purpose of prevention and treatment.

Specifically, the present invention provides a composition for the treatment of Alzheimer’s disease, characterized in that it comprises β-nicotinamide mononucleotide (NMN) and L-α-glycerophosphorylcholine (α-GPC), wherein β-nicotinamide mononucleotide is used in a dose of 3 to 9 mg/kg/d and L-α-glycerophosphorylcholine is used in a dose of 8 to 20 mg/kg/d.

As a preferred embodiment of the present invention, the pharmaceutical formulation can be a tablet or capsule.

The present invention also provides a functional food product comprising the above-mentioned composition for the treatment of Alzheimer’s disease and an edible excipient.

The compositions of the present invention can be applied in the preparation of drugs for the treatment of Alzheimer’s disease.

NMN (Nicotinamide mononucleotide) in the present invention is short for β-nicotinamide mononucleotide, naturally present in living cells, and is a precursor of coenzyme 1 NAD+ (nicotinamide adenine dinucleotide). It may extend the life span appropriately. The real anti-aging effect is an important energy metabolite called nicotinamide adenine dinucleotide (NAD+). NMN is a nutrient that is commercially available commercially. NMN is converted to NAD+ in cells, it is classified as a new type of vitamin B3 and goes beyond the effects of ordinary vitamins and is considered a preventive and rejuvenating supplement for aging. NMN itself is a substance contained in the human The body contains substances that are present in breast milk and in foods such as mauve beans, broccoli, cucumbers, cabbage, avocados, tomatoes, etc., but in very small amounts (0.25-1.88 mg per 100 g). Raw beef and shrimp, etc. also contain very small amounts (0.06-0.42 mg per 100 g) of NMN components. So to boost NMN, you also have to supplement directly from outside. David Sinclair, a Harvard professor of genetics, suggests that NMN has the effect of reversing aging: by maintaining sufficient NAD+ in cells, the self-repairing ability of DNA can be maintained, so that the DNA damage caused by aging can be effectively repaired, thus inhibiting aging. The raw material of NMN is the precursor of NAD+, and taking NMN can increase the level of NAD+ in the body. However, to date, there is no direct evidence to confirm that NMN intake can treat or improve the symptoms of Alzheimer’s disease.

The α-GPC in this invention is the abbreviation of L-α-glycerophosphorylcholine, which is a water-soluble substance with 2 fatty acids removed from the main phospholipid PC (phosphatidylcholine) that constitutes the cell membrane and is originally widespread in living organisms and is one of the body components present in human breast milk and body fluid swelling. In the 2009 revision of the food and drug classification, L-α-glycerophosphorylcholine (sometimes called sn-glycerophosphorylcholine) was defined as food. α-GPC is one of the few nutrients that can cross the blood-brain barrier, so it can be rapidly converted from choline to acetylcholine, and is considered to be a nutrient that prevents choline deficiency, increases neurotransmitters, and promotes the secretion of growth hormone.

Moreover, since acetylcholinesterase inhibitors (AChEI) are by far the most widely used drugs for the treatment of AD in clinical practice, among the 5 drugs approved by the FDA for the treatment of AD, except for memantine which is an NMDA receptor antagonist, the remaining 4 [Tacrine, Donepezil, Galanthamine and Rivastigmine ] are all AChEI. However, there are no reports so far that α-GPC intake can treat or improve the symptoms of Alzheimer’s disease, probably because α-GPC intake alone does not increase the acetylcholine content due to the body’s modulation.

However, the inventors of the present invention unexpectedly found that the use of a combination of two endogenous nutrients, NMN and α-GPC, showed symptom amelioration in animal models of Alzheimer’s disease, suggesting the promise of NMN and α-GPC as prevention or treatment of Alzheimer’s disease, as can be seen in the specific embodiments in the Examples.

As a preferred embodiment of the present invention, the present invention also provides a pharmaceutical formulation for the treatment of Alzheimer’s disease comprising the composition described above, and further comprising pharmaceutically acceptable excipients.

The compositions of the present invention are preferably made into solid dosage forms for oral administration. Solid dosage forms for oral administration include capsules, tablets, pills, bulk, and granules. In these solid dosage forms, the composition is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with

  • a filler or bulking agent, e.g., starch, lactose, sucrose, glucose, mannitol, and silicic acid;
  • (b) a binder, e.g., hydroxyethylcellulose, alginate, gelatin, polyvinylpyrrolidone, sucrose, and gum arabic;
  • (c) humectants, e.g., glycerin;
  • (d) disintegrants, e.g., agar, calcium carbonate, potato starch or tapioca starch, alginate, certain complex silicates, and sodium carbonate;
  • (e) retardants, e.g., paraffin;
  • (f) absorption accelerators, e.g., quaternary amine compounds;
  • (g) wetting agents, e.g., cetearyl alcohol and glycerol monostearate;
  • (h) adsorbents, e.g., kaolin; and
  • (i) lubricants, e.g. talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium dodecyl sulfate, or mixtures thereof. In capsules, tablets, and pills, the dosage form may also contain a buffering agent.

Solid dosage forms such as tablets, sugar pills, capsules, pills and granules may be prepared using coatings and shell materials such as enteric coatings and other materials well known in the art. They may contain opaque agents and the release of the active compound in such compositions may be delayed in a portion of the digestive tract. Examples of embedding components that may be used are polymeric substances and wax-like substances. If necessary, the composition may also be formed in microcapsule form with one or more of the above excipients.

Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the compositions of the present invention, liquid dosage forms may contain inert diluents routinely employed in the art, such as water or other solvents, solubilizers, and emulsifiers, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, and sesame oil, or mixtures of these substances. In addition to these inert diluents, the compositions may also contain additives such as wetting agents, emulsifiers and suspending agents, sweeteners, flavor correctors, and fragrances.

In addition to the compositions of the present invention, suspensions may comprise suspending agents, such as, for example, ethoxylated isooctadecanol, polyoxyethylene sorbitol, and dehydrated sorbitol esters, microcrystalline cellulose, methanolic aluminum, and agar, or mixtures of these substances. Compositions for parenteral injection may comprise physiologically acceptable sterile aqueous or anhydrous solutions, dispersions, suspensions or emulsions, and sterile powders for redissolution into sterile injectable solutions or dispersions. Suitable aqueous and non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

From the point of view of ease of administration, the compositions of the present invention are preferably made into tablets or capsules. In order to prepare tablets or capsules of the powders of the compositions of the present invention, the powders of the compositions of the present invention may be mixed with a formulation carrier, which is a commonly used tablet-making component such as corn starch, sucrose, sorbitol, fatty acids, magnesium stearate, and other pharmaceutical diluents.

The compositions of the present invention for the treatment of Alzheimer’s disease can be used in combination with other classes of drugs for the treatment of Alzheimer’s disease. As other drugs for the treatment of Alzheimer’s disease, tacrine, donepezil, carboplatin, galantamine hydrobromide, memantine, nine-phase I, etc. may be cited as clinically common drugs.

The present invention also provides a functional food product comprising the above-mentioned compositions for treating Alzheimer’s disease and edible excipients.

Since the composition of the present invention is fully edible due to the raw materials, it is well suited for the development of functional food products, such as functional food products that can be prepared in the form of oral tablets, confectionery, solid drinks, liquid drinks. The functional food product of the present invention, comprising the above composition of the present invention and edible excipients, as a food additive that can be used in compliance with relevant standards, such as food additives in compliance with GB2760-2014 of the People’s Republic of China.

As a preferred form of the functional food of the present invention, oral tablets can be cited. There is no special limitation on the way of making the composition of the present invention into oral tablets, and a well-known method can be used, for example, the process of granulating the composition of the present invention, mixing it with edible excipients, etc., and carrying out compression of the tablets. As edible excipients, starch, sucrose, methylcellulose oligofructose, tea powder, salt, stevioside, sorbitol, vitamin C, etc. can be cited, but not limited to these, as long as the excipients conforming to food standards (e.g. GB2760-2014 of the People’s Republic of China) can be added appropriately.

The raw material of the invention itself has no irritating taste, and when it is made into oral tablets, it will not have an undesirable taste and does not need to be coated. However, it can be coated if it is for the purpose of facilitating consumption, facilitating prevention of moisture absorption, etc. From the food point of view, it is preferred to use excipients such as sodium alginate, xanthan gum, gum arabic, etc. that are fully compliant with food standards for coating.

As a preferred form of functional food of the present invention, the composition of the present invention, from the viewpoint of strong food-oriented appearance and high acceptability, can also be made in the form of liquid for oral consumption of food. The dosage forms of liquid formulations for oral administration as food are solutions, syrups, or suspensions. In the preparation of these liquid formulations, the dispersing medium is generally water, non-water-soluble excipients such as almond oil, oils, and fats; fillers allowed as pharmacological include such as sorbitol, hydrogenated edible oil, methyl cellulose; also preservatives such as methyl or propyl P-phenol; emulsifiers such as lecithin, arabinoxylan: artificial colors or sweeteners.

As the dose to be administered, it should depend on the patient’s age, weight, health status and dosage form. The recommended doses are 3 to 9 mg/kg/d for β-nicotinamide mononucleotide and 8 to 20 mg/kg/d for L-α-glycerophosphorylcholine. However, since the raw material components of the present invention all have a strong safety profile, a person skilled in the art can increase the effective dosage of each component as appropriate for individual needs.

The inventors of the present invention have verified the efficacy of the compositions of the present invention for the treatment of Alzheimer’s disease through experimental animal models. Specifically, the Morris water maze experiment was used in this study, which is an experiment that forces experimental animals to swim and learn to find a platform hidden in the water, and is mainly used to test the learning memory ability of experimental animals for spatial position sense and direction sense (spatial orientation), and is widely used in the pharmacological efficacy evaluation studies of Alzheimer’s disease.

In the present invention, an animal model of Alzheimer’s disease was prepared by using the classical Aβ1-40 for directional injection into the hippocampal CA1 region of rats, combined with the classical Morris water maze to test the learning and memory behaviors of the model rats, so as to evaluate the effect of the administered composition. The results showed that rats gavaged with NMN alone did not show improvement in Alzheimer’s disease symptoms, and similarly, rats gavaged with α-GPC alone did not show improvement in Alzheimer’s disease symptoms. However, when the combination of NMN powder and α-GPC was used for gavage, it produced cognitive improvement in the model rats, suggesting that the combination of NMN and α-GPC has promising applications for the prevention or treatment of Alzheimer’s disease.

The composition of the present invention has the following obvious advantages over the prior art: the raw materials of the composition developed by the present invention are all foodstuffs, which are combinations with clear effects and convincing safety for the treatment of Alzheimer’s disease, the composition of the present invention and using scientific animal phenotypes to verify the exact effect, although its effect is weaker than the efficacy of the western drug according to the control, considering its special suitability to meet can It is suitable for long-term use and can be used to prevent Alzheimer’s disease symptoms, and therefore has a broad market prospect.

Description of the accompanying figures

  • 1 is a statistical chart of the evasion latency results for each group of rats in Example 2.
  • 2 is a statistical chart of the results of the platform penetration test for each group of rats in Example 2.

Specific embodiments

The following collection of specific embodiments describes the present invention in further detail. The embodiments are intended only to illustrate examples of the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention in any way.

Example 1 Establishment of an experimental animal model of AD

The model animal strain was selected from SPF-grade SD rats, healthy, male, weighing 250g-300g, and provided by Shanghai Southern Model Biotechnology Development Co. The experimental groups were divided into normal control group, AD model group, positive drug group, test drug group 1, test drug group 2, and test drug group 3, a total of six groups, with 8 to 11 animals in each group.

Except for the normal control group, β-amyloid (specifically Aβ1-40, purchased from Sigma) injection into the hippocampus was used to induce rats as AD rats in a modeling approach. Specifically, the rats were anesthetized by intraperitoneal injection of 10% chloral hydrate solution at a dosage of 4.5 ml/kg. SD rats were all anesthetized by intraperitoneal injection of 1% sodium pentobarbital 40 μg/g, followed by fixation on a brain stereotaxic apparatus after anesthesia. According to the brain stereotaxic atlas of rats, the zero starting point was bregma, the puncture point was 3.5 mm after bregma, the right side of the midline was opened by 2 mm, and then the skull was drilled with a dental drill, and a microinjector was used to inject 10 μg (1 μL) each of Aβ1-40 into the CA1 area of the hippocampus bilaterally at a slow and uniform rate, and the needle was left for 5 min. After 7 d of post-modeling recovery, gavage feeding was started with saline in the normal control group and AD model group, meperidine hydrochloride 3 mg/kg/d in the positive drug group, β-nicotinamide mononucleotide 8 mg/kg/d in the test drug group 1, and L-α-glycerophosphorylcholine 10 mg/kg/d in the test drug group 2. 10 mg/kg/d, subject group 3 gavaged β-nicotinamide mononucleotide 8 mg/kg/d and L-α-glycerophosphorylcholine 10 mg/kg/d, and immediately after 21 days of continuous administration, the following four days of behavioral tests were performed.

Example 2 Behavioral assay – Morris water maze model test

The escape latency test was performed on the first 3 days, and the platform was removed for the platform penetration test on the 4th day.

2.1 Evasion latency test

The so-called latency period experiment refers to each rat test from the same position into the water, there must be experimental personnel to assist the rat slowly into the water. In order to ensure that the rats swim on the platform environment as much as possible the same, pay attention to the replenishment of water to maintain the platform from the surface of the water 2cm, the rat into the water and start timing, find the platform and stay for 2 seconds, record the time, the time spent for the latency period, more than 120 seconds still have not found the platform is also recorded as 120 seconds. The shorter the time indicates the better the memory improvement effect.

2.2 Platform penetration experiment

The so-called stage crossing experiment refers to the removal of the platform in the water maze after the first 3 days of the water maze latency test, and record the movement trajectory of the rats after entering the water maze, and once intersecting with the platform position is recorded as one stage crossing, and the total number of stage crossing for each rat within 120 seconds is recorded.

2.3 Results and analysis

Each rat was put into water 4 times in each quadrant on the first 3 days, and a total of 12 times were dropped into water. The final time values obtained were summed up and averaged and rounded off as the final latency of the rat, and the 4th day each rat was put into water 4 times in each quadrant, and the 4 numbers obtained were averaged as the number of times the rat wore the stage.

Using statistical analysis, the latency to the stage and the number of stage penetrations were compared between the model group and the normal group. In the present invention, the normal control group and the AD model group were statistically different in latency and number of stage penetration, and the modeling was considered successful (p<0.05 was considered statistically different, and p<0.01 was considered statistically significant). The experimental results showed that the model of the invention was usable at p<0.05 between the normal control group and the AD model group.

Using statistical analysis, the model group was compared with each administered subgroup for stage latency and the number of stage penetration.

  1. The effect of each group on the latency of evasion in model rats

The positive control group (meperidine group) showed a more pronounced efficacy, almost at the same level as the normal control group. Test drug group 1 and test drug group 2 did not show efficacy compared to the AD model group, but test drug group 3 showed a significant effect on memory improvement compared to the AD model, although not as pronounced as the positive control group, but its efficacy was significantly different (p<0.05).

Table 1 Statistical table of the latency period of evasion in rats

Statistical table of the latency period of evasion in rats

  1. Effectiveness of each group on the rat table-penetration test

Similar to the above-mentioned evasion latency, test drug group 1 and test drug group 2 did not show efficacy compared to the AD model group, but test drug group 3 showed a significant effect on memory improvement compared to the AD model, but its efficacy was significantly different (p<0.05). The positive control group (meperidine group) showed less efficacy than the escape latency statistic, but it also showed clear efficacy.

Table 2 Statistical table of the rat penetration table test

Statistical table of the rat penetration table test

The animal experiments described above showed that the compositions used in subject group 3 showed promising applications in the treatment of Alzheimer’s disease, i.e., the compositions of the present invention have therapeutic or preventive activity against Alzheimer’s disease and can be used in the preparation of drugs or functional foods for the treatment and prevention of Alzheimer’s disease.

Finally, it should be noted that the above-described embodiments are used only to illustrate the technical solutions of the present invention and not to limit them; despite the detailed description of the present invention with reference to the preceding embodiments, it should be understood by those of ordinary skill in the art that it is still possible to modify the technical solutions recorded in the preceding embodiments or to make equivalent substitutions of some or all of the technical features thereof; and that these modifications or substitutions do. These modifications or substitutions do not take the essence of the corresponding technical solutions out of the scope of the technical solutions of the embodiments of the present invention.

All publications and patent documents cited in this specification are incorporated herein by reference as if each publication or patent were separately and expressly identified as being incorporated herein by reference. Various changes and replacements with equivalents may be made to the various embodiments disclosed herein without departing from the substance and scope of the present application disclosure. Unless the context indicates otherwise, any feature, step, or embodiment of an embodiment of the present disclosure may be used in combination with any other feature, step, or embodiment.

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