Triple pharmaceutical composition for proteinaceous infectious diseases

Abstract

Methods for treating and preventing prion protein complex infections are disclosed. Transcription of the amyloid precursor protein gene and PrP gene and RNA transcripts are rate-limiting steps in amyloid protein formation and PrP. sc It is most feasible to interrupt and control the process of formation. Therefore, treatment and prevention of prion protein complex infections will interrupt this process at the level of transcription of DNA into RNA, transport of RNA to mitochondria for protein synthesis, and deposition in cerebral cortical neurons.

Description

[Technical field]

[0001] The present disclosure relates to the treatment of Alzheimer's disease (AD). [Background technology]

[0002] AD is generally considered to be a localized disease of the brain. Neurological AD is the third leading cause of death in the United States, after cardiovascular disease and cancer. AD usually follows a sequence of neuroinflammation, amyloid and tau proteopathy, accumulation deposition, neurotoxicity and neuronal damage, loss of function (i.e., activities of daily living (ADL) and cognitive abilities), and finally death. Death from AD is due to the futility and loss of will to live of these patients who have become depersonalized and have lost their appetite for food and liquids, which, combined with growth failure, results in premature death, usually within 5 to 10 years of AD diagnosis.

[0003] There are four main prevailing theories regarding the causes of AD: (a) the cholinergic hypothesis, (b) the amyloid protein deposition hypothesis, (c) the tau protein deposition hypothesis, and (d) the neurovascular hypothesis. Currently, there are no effective treatments that can modulate the progression of Alzheimer's disease or prevent its onset. Currently available treatments only work to improve symptoms, while the development of treatments that can halt or slow the progression of the disease remains a challenging unmet need.

[0004] According to the cholinergic hypothesis, degeneration of cholinergic neurons in the basal forebrain and the associated loss of cholinergic neurotransmission in the cerebral cortex and other regions contributes significantly to the cognitive deterioration seen in patients with Alzheimer's disease.

[0005] The amyloid protein deposition hypothesis posits that the formation of amyloid plaques and neurofibrillary tangles contributes to the deterioration of neurons (nerve cells) in the brain and the subsequent symptoms of Alzheimer's disease. Amyloid proteins are a large group of proteins with as many as 60 reported types. Thirty-six amyloid proteins have been linked to human disease. Amyloid proteins were first discovered and described by Rudolf Virchow, who considered them to be starchy, and hence came to be called starch-related amyloid, or "amylin" in Latin. Amyloid proteins were then thought to be fatty substances, but were later found to be proteinaceous substances. Since the introduction of sophisticated protein chemistry, mass spectrometry, and X-ray crystallography, amyloid proteins have been better characterized and have been identified in a variety of human diseases and conditions.

[0006] Amyloid protein diseases were previously classified into primary and secondary amyloidosis. Primary diseases were recognized as familial disorders involving protein synthesis and deposition in organs such as the heart, kidney, skin, tongue, adipose tissue, and rectum. In secondary diseases, amyloid protein diseases were recognized as secondary to chronic suppurative diseases such as tuberculosis or other bacterial abscesses, which are prevalent in developing and underdeveloped countries around the world. Similarly, chronic inflammatory conditions such as rheumatoid arthritis and renal dialysis also lead to secondary, reactive amyloid protein deposition.

[0007] Amyloid precursor protein (APP), encoded by chromosome 21, is implicated in AD. APP is a transmembrane protein that spans neuronal membranes and is important for neuronal growth, survival, and repair after injury. Thus, loss of neuronal APP may affect physiological and pathophysiological defects that lead to dementia. Clinical data from individuals with Down syndrome (trisomy 21) show that because the APP gene is located on chromosome 21 and carries three copies, they develop AD earlier, in their 30s and 40s. This is similar to how inflammatory bowel disease (IBD) patients develop colon cancer in their 30s and 40s compared to healthy individuals who develop colorectal cancer in their 50s and 80s. APP is copied and used to synthesize amyloid proteins.

[0008] Amyloid-β (Aβ) is the specific amyloid protein involved in AD. Amyloid plaques are made of small peptides 39-43 amino acids in length. Amyloid-β is generated from sequential cleavage of APP by β-site amyloid precursor protein-cleaving enzyme 1 (BACE-1) followed by γ-secretase. In AD, γ-secretase and β-secretase act together in a proteolytic catabolism reaction to cleave smaller fragments of APP. These proteolytic fragments then form amyloid-β fibrils and further form clumps deposited outside neurons known as senile plaques.

[0009] Because Aβ accumulates in excess in AD, it is logical to infer that its precursor, APP, would be elevated as well. However, one study showed that neuronal cell bodies contain less APP in response to their proximity to amyloid plaques. It has been hypothesized that this lack of APP near Aβ plaques is due to reduced production of APP, which is normally elevated in response to stress.

[0010] Several BACE-1 inhibitors, BACE-2 inhibitors, and humanized monoclonal antibodies against soluble amyloid proteins have been used in clinical trials for AD. These trials have failed to realize the promise of being disease-modifying (DMD) agents in AD (i.e., they change the underlying pathology of the disease). Similarly, vaccines have also been tried to remove amyloid protein plaques in AD, but all to no avail. Given the failure of clinical trials with BACE inhibitors and the failure of amyloid immunotherapy with intravenous solanezumab, the amyloid protein deposition hypothesis has come into question.

[0011] The tau deposition hypothesis proposes that abnormalities in tau protein initiate the disease cascade. In this model, hyperphosphorylated tau starts to pair with other tau threads. Eventually, they form neurofibrillary tangles inside the neuronal cell body. When this happens, microtubules break down and the structure of the cell's cytoskeleton is disrupted, which disrupts the neuronal transport system. This can result first in failure of biochemical communication between neurons and then in cell death.

[0012] The neurovascular hypothesis proposes that a significant amount of Aβ peptide in the brain of Alzheimer's disease patients originates from the systemic circulation. According to this theory, a compromised blood-brain barrier (BBB) ​​is involved. One side effect of this compromise is the production of amyloid and hyperphosphorylation of tau.

[0013] Prions (PrP) are proteins that arise from the misfolding of normal proteins. There are two forms of prions: the normally folded protein PrP c and the disease-causing misfolded form, PrP sc The two forms do not differ in amino acid sequence, however, pathogenic PrP sc The isoforms are PrP with normal secondary and tertiary structures. c The morphology is different from that of PrP. scisoform is rich in β-sheets, whereas normal PrP c The morphology is rich in α-helices. This difference in the three-dimensional structure is what causes PrP sc PrP associates with amyloid-β fibrils and becomes extremely resistant to proteolysis by enzymes or other chemical, radiological and physical means, whereas the normal form is susceptible to complete proteolysis and is soluble in non-denaturing detergents. sc is PrP c From PrP sc It can promote the conversion of PrP sc Other PrP c This causes a chain reaction, which then multiplies rapidly, resulting in the onset of prion disease. c The protein is one of several intracellular receptors for soluble amyloid β protein (Aβ) oligomers.

[0014] The highly conserved genetic sequence of PrP, but without a distinctive function, has puzzled researchers for many years. Researchers have tried to link PrP and Aβ to other diseases without success. Some studies have tried to hypothesize that Aβ must also be associated with heart disease because Aβ is a type of protein plaque. However, some heart diseases do not contain Aβ plaques, and a significant number of AD patients do not suffer from heart disease. Studies of treating heart disease with antibiotics have not been successful. Not only have these studies focused on treating bacteria, rather than inflammation or plaque accumulation, but these studies have concluded that antibiotic treatment is unlikely to have a beneficial effect. One study summarized the current state of the field as follows: "Clinical trial data to date do not provide sufficient support for the clinical use of antibiotics in the primary or secondary prevention of coronary artery disease" [Anderson et al., 2004].

[0015] Apolipoprotein E (ApoE) transports lipids, fat-soluble vitamins, and cholesterol to the lymphatic system and then to the blood where it is the major cholesterol transporter in the brain. The gene for ApoE maps to chromosome 19. ApoE is polymorphic with three major alleles, ApoE-ε2, ApoE-ε3, and ApoE-ε4. These allelic forms differ from each other by only one or two amino acids at certain positions, but these differences alter the structure and function of ApoE. These differences have physiological effects. ApoE-ε4 has an allelic frequency of approximately 14% and is believed to be significantly involved in late-onset Alzheimer's disease (LOAD).

[0016] Research into treatments for AD has also not produced promising results. Researchers have tried a range of potential treatments, from designer drugs to custom proteins to antibiotics, but none have been successful. Numerous studies suggest that antibiotic treatments cannot cure AD. In 2013, researchers conducting a study on doxycycline and rifampin stated, "Conclusion: 12 months of treatment with doxycycline or rifampin, alone or in combination, has no beneficial effect on cognition or function in AD" [Molloy et al., 2012].

[0017] Another study looking at potential treatments for AD characterized the Molloy study as follows: "However, there were no differences between the treatment and placebo groups at 3 and 12 months. However, no beneficial effects on cognition or function were detected when doxycycline was used alone or in combination with rifampin" [Appleby et al., 2013].

[0018] Other studies treating AD patients with antibiotics have shown that both treated and untreated groups experienced declines in cognitive function [Boyles, 2003]. AD researchers are skeptical of antibiotic regimens as a treatment for AD, with one stating, "We do not propose that long courses of antibiotics would be curative -- that would be unreasonable for a number of reasons" [Minter, 2016].

[0019] Researchers are also skeptical that other drugs can treat AD. The National Institute on Aging states that NSAIDs work primarily by targeting inflammation, but to date, there have been no clinical trials in which AD patients have benefited from the actual use of NSAIDs. NSAIDs have been considered poor candidates for AD for many reasons. One reason is that AD often develops in older patients, who generally have weaker hearts. In fact, a study to see if NSAIDs could be used to combat AD had to be halted due to concerns about the cardiovascular health of participants [Terrie, 2009]. Acetaminophen and aspirin were found to have no beneficial effect on AD in one study [Szekely et al., 2008]. In another study, naproxen and celecoxib did not improve cognitive function in AD patients [Martin et al., 2007].

[0020] In this context of prion protein complex infections, the present inventors focus on several drugs that have not been proposed for therapeutic application against prion protein complex infections. For example, although engineered antibodies have been tried, antibiotics have not been considered as possible treatments against prion protein complex infections. Another class not previously considered is immunosuppressants.

[0021] Tetracyclines are a very old group of bacteriostatic antibiotics consisting of tetracycline, doxycycline and minocycline. They act by inhibiting protein synthesis in bacterial and protozoan cells and in eukaryotic mitochondria, thereby inhibiting the binding of aminoacyl-tRNA to the mRNA ribosomal complex. Tetracyclines exert their function mainly by binding to the 30S ribosomal subunit in the mRNA translation complex. In addition to inhibiting protein synthesis, these drugs are anti-inflammatory, lipid soluble and have high concentration in the central nervous system.

[0022] Sirolimus, also known as rapamycin, is a macrolide compound sold by Pfizer under the trade name Rapamune®. There are numerous derivatives of rapamycin (rapalogs) that function as mTOR inhibitors. New rapalogs are being researched and developed daily. Sirolimus has immunosuppressive effects in humans and is used to prevent kidney transplant rejection. Sirolimus inhibits the activation of T and B cells by reducing sensitivity to interleukin 2 (IL-2) through mTOR inhibition. Sirolimus prevents the humoral immune system from synthesizing humoral antibodies against kidney grafts by its effect on B cells. Sirolimus and its rapalogs increase autophagy and mitophagy. Summary of the Invention [Means for solving the problem]

[0023] The present invention provides methods for treating and preventing prion protein complex infections. [Brief description of the drawings]

[0024] [Figure 1] Figure 1 is the skeletal formula of tetracycline with the atoms and the four rings numbered and labeled. [Diagram 2] Figure 2 is the formula for doxycycline. [Diagram 3] FIG. 3 is the formula for minocycline. [Figure 4] FIG. 4 is the formula for sirolimus. [Diagram 5] FIG. 5 is a schematic diagram showing the basis for the exponential increase in the presence of Aβ and PrPsc. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0025] AD is not a localized disease of the brain. AD is a prion protein complex infection, a systemic disease involving both the systemic and peripheral circulation and B cells. AD involves a localized reaction in the neocortex. Indeed, evidence for this is the high expression of Aβ in saliva. 42 Levels (by ELISA test) of Aβ in blood 42 / 40 Ratio and Aβ in cerebrospinal fluid 42 In addition, AD can also be initiated by brain inflammation caused by other diseases such as Parkinson's disease and chronic traumatic encephalopathy (CTE) caused by repeated brain trauma in boxers and football players.

[0026] The deposition of amyloid-β protein seen in AD is secondary to a chronic neuroinflammatory state in acetylcholine-excreting neurons of the cerebral cortex. This amyloid protein deposition begins 10-15 years before the clinical diagnosis of AD in patients and continues until the patient's death. Transcription and RNA transcription of the APP gene are the rate-limiting steps, and it is most feasible to block and control the process of amyloid protein formation. Thus, described or proposed herein is a treatment for AD based on interrupting this process at the level of transcription of DNA into transcriptional RNA (tRNA), messenger RNA (cmRNA), RNA transport to mitochondria for protein synthesis, and deposition in cerebral cortical neurons. This is the gist of our efforts to introduce the first DMD of AD.

[0027] This neuroinflammation in the neocortex is mediated by the local secretion of amyloid beta into memory nerve fibers that secrete acetylcholine, and the secretion of cellular prion protein (PrP c ) peptide and tau peptide secretion. Neurotoxicity of amyloid protein oligomers leads to the secretion of PrP c The peptide changes from an α-helical structure to a β-helical structure (i.e., PrP sc ) is misfolded. scThe β-helical structures of Aβ interact with Aβ fibrils and initiate the laying down of sheets of Aβ fibrils that are neurotoxic, causing neurotoxicity and death of neurofibrils and neurons, and producing the amyloid plaques and tau protein tangles that are characteristic of the disease.

[0028] Figure 5 shows Aβ and PrP in AD. sc As shown in Figure 5, the presence of PrP sc PrP c When applied to PrP c is PrP sc A similar behavior occurs with APP and Aβ, where Aβ seeds APP for the production of more Aβ. However, these two cycles are not independent. They are interdependent: Aβ misfolds into PrP c From PrP sc This serves as a seed for the conversion to PrP sc APP is the seed that produces Aβ. Thus, Aβ and PrP are at the center of the prion protein complex in the amyloid plaques characteristic of AD. sc The witch's elixir is PrP c From PrP sc This leads to a deadly cycle that cannot be stopped by therapies that only inhibit misfolding into APP or by the production of Aβ from APP. The therapies described herein address both types of misfolding and the production of a single precursor protein (or oligopeptide).

[0029] AD may be treated and prevented by two forms of therapy. Amyloid-β protein present in the blood diffuses into the cerebrospinal fluid that flows into the brain and neocortex. This results in a secondary neocortical response in which sheets of amyloid-β fibrils are laid down, causing the death and destruction of memory cells, leading to amyloid plaques and tangles of neurofibrillary tau protein. One form of therapy uses immunosuppressants to address the systemic humoral B-cell response and the transcription, translation and synthesis of prion protein. The other form of therapy uses antibiotics to address the synthesis of amyloid-β protein. Combinations of these forms of therapy can be effective.

[0030] AD arises from a complex of rogue prion proteins - the witch's elixir. This complex of rogue prion proteins consists of Aβ fibrils and prion receptor protein (PRP)β (PrP sc ) fibrils. The body's natural response to the misbehaving prion protein complexes is a self-defense mechanism that harms the tissues itself. These defense mechanisms take the form of self-assembling Pacman that attacks and eats the misbehaving prion protein complexes. The corresponding damage to tissues is the culprit in the pathogenesis of AD and other prion protein complex infections.

[0031] The systemic disease component of AD may be treated with immunosuppressants such as sirolimus. Sirolimus, by its effect on B cells, impairs the humoral immune system from synthesizing humoral antibodies and APP. This impairs the systemic component of AD pathogenesis. Currently, sirolimus is only prescribed for patients undergoing transplants and certain cancers. The combination of sirolimus with tetracycline is off-label. Sirolimus also inhibits the formation of antibodies and generally inhibits the synthesis of amyloid beta protein by plasma cells and B cells. Sirolimus also suppresses the innate immune system and cytokines produced by microglia and macrophages, such as TNFα, IL-1β, IL-6, and gamma interferon.

[0032] The central nervous system (CNS) localized effects of AD may be treated with antibiotics such as tetracyclines, which block protein synthesis through their effects on transcription, translation, and binding to the ribosomal protein complex. Tetracycline compounds can address the CNS / neocortex component of AD pathogenesis by inhibiting the transcription of the APP gene on chromosome 21 and the transcription of the PrP gene on chromosome 20. In addition, tetracyclines block gene translation and protein synthesis by binding to the 30S and 50S subunits of the ribosomal protein complex.

[0033] The dual action of treating both the systemic disease component and the CNS localized effects leads to the inhibition or abolition of the effects of the malformed prion protein complex in AD. By inhibiting transcription and blocking synthesis of amyloid protein in AD patients, further deposition of amyloid protein in the cerebral cortex and subsequent neurotoxicity and neuronal damage, as well as loss of memory and function, can be prevented. Accordingly, patients regain function and are able to participate in activities of daily living and in interactions with their families. Similarly, PrP sc By blocking the transcription and synthesis of the prion protein, the second part of the misbehaving prion protein complex is destroyed.

[0034] AD patients may be characterized as three types. In type I, patients have Down syndrome, which accounts for approximately 0.001% of all AD cases. In type I patients, AD manifests at approximately 30-40 years of age. The genetic marker is Trisomy 21. In type II, patients have early onset Alzheimer's disease (EOAD), which accounts for approximately 1-5% of all AD cases. In type II patients, AD manifests at approximately 50-60 years of age. The genetic markers are PSEN1 on chromosome 14 and PSEN2 on chromosome 1. In type III, patients have late onset Alzheimer's disease (LOAD), which accounts for approximately 95% of all AD cases. In type III patients, AD manifests at approximately 65-90 years of age. Two-thirds of type III patients are APOEε4 positive, with APOEε4 present on chromosome 19. Type III patients may be further differentiated as either APOEε4 positive or APOEε4 negative.

[0035] Type III APOE ε4 positive produces a defective apolipoprotein E (ApoE) protein. This defective protein makes it difficult for the body to metabolize proteins, fats, and lipids, and makes it difficult for the body to deliver nutrients and cholesterol to the brain. Thus, APOE ε4 positive patients are hit harder when they develop AD compared to type III AD patients whose APOE is still functioning (APOE ε4 negative).

[0036] Appropriate treatment may vary depending on the characteristics of AD a patient has. All types may benefit from a combination of an antibiotic such as tetracycline and an immunosuppressant such as sirolimus. Type III patients who are APOE ε4 positive should also receive a lipophilic statin such as Atovar (atorvastatin), which crosses the blood-brain barrier.

[0037] The dosage may be in the form of a unit dosage, i.e., a single pill, tablet or capsule.

[0038] For adults, an appropriate therapy may be one of the following: (a) 50 mg tetracycline, 1 mg sirolimus, 10 mg atorvastatin; (b) 50 mg tetracycline, 1 mg sirolimus, 20 mg atorvastatin; (c) 100 mg tetracycline, 1 mg sirolimus, 40 mg atorvastatin; (d) 100 mg minocycline, 1 mg sirolimus, 80 mg atorvastatin; (e) 50 mg tetracycline, 2 mg sirolimus, 10mg atorvastatin; (f) 50mg tetracycline, 2mg sirolimus, 20mg atorvastatin; (g) 100mg tetracycline, 2mg sirolimus, 40mg atorvastatin; (h) 100mg minocycline, 2mg sirolimus, 80mg atorvastatin; (i) 2mg sirolimus, 100mg minocycline and 20mg atorvastatin; (i) 2mg sirolimus, 100mg minocycline and 20mg atorvastatin. Note that the dose of atorvastatin may be given twice a day, for example, to achieve a dose of 20mg, a patient may take 10mg atorvastatin in the morning and 10mg atorvastatin at another time. The tetracycline may be either minocycline or doxycycline. Other oral or intravenous tetracyclines may also be used. Simvastatin and lovastatin, both of which are lipophilic, may also be used.

[0039] Physicians who prescribe this triple approach should be aware of the effect of statins on the patient's cholesterol and triglyceride levels. In instances where atorvastatin should not be used, it may be replaced with another statin as listed above. The dosage of the new statin may be the same as atorvastatin or adjusted based on the effect of the statin. For patients with high cholesterol or heart disease, the dosage of statin or atorvastatin may be further adjusted slightly to take into account the patient's blood chemistry and neurological effects.

[0040] When manufacturing a triple-drug combination that includes minocycline or doxycycline, sirolimus, and atorvastatin, it may be necessary to manufacture different drugs with different atorvastatin dosages and distribute them to patients with certain diseases.For example, when manufacturing pills that include three drugs, one pill may contain 50 mg minocycline, 1 mg sirolimus, and 40 mg atorvastatin for the majority of the population that suffers from AD, but another pill may be required that contains 50 mg minocycline, 1 mg sirolimus, and 80 mg atorvastatin for patients that already suffer from high cholesterol and AD.In another example, a pill with a lower atorvastatin dosage, such as 50 mg minocycline, 1 mg sirolimus, and 20 mg atorvastatin, may be manufactured for patients that already take another dose of atorvastatin or another statin.

[0041] The effects of the therapies described herein may be evident in as little as 72 hours. At other times, effects may be seen in 3 to 12 months. If the treatment is effective, the patient may discontinue treatment with controlled observation for possible relapse and retreatment, or may continue treatment at a maintenance dose. For AD patients, the effects may be measured by the Alzheimer's Disease Assessment Scale-Cognitive (ADAS-Cog) subscale and the Alzheimer's Disease Cooperative Study-Activities of Daily Life (ADCS-ADL) scale or the Mini-Mental Status Examination (MMSE). Both of these tests have been developed over many years and will likely continue to be improved in the future.

[0042] The frequency with which the medication is taken may vary, as well as the time of day. Every day or every other day is sufficient for some patients, while others may take it for 3 days and rest for 2 days. These are examples of drug-free periods. The dosage may vary from day to day. The time of day to take the medication may be chosen based on the patient having an empty stomach for better absorption.

[0043] The drugs in the therapy may be administered together, separately, or at least one separately and at least some together. The drugs in the therapy may be administered orally (e.g., tablets), topically (e.g., patches), intranasally (e.g., inhalation), or parenterally, such as by immediate release formulation, or by a sustained release formulation, an extended half-life injectable formulation, or at least one sustained release formulation and at least one extended half-life injectable formulation.

[0044] Dosage levels may vary, with daily doses of doxycycline as low as 40 mg, minocycline as low as 25 mg, and sirolimus as low as 0.5 mg. At the higher end, dosages may be 400 mg doxycycline (e.g., 200 mg twice daily), 300 mg minocycline (e.g., 150 mg twice daily), and 4 mg sirolimus (e.g., 2 mg twice daily). The dosages above are for an average adult, and dosages may be correlated with body weight, with heavier patients receiving larger doses and lighter patients receiving smaller doses. Dosages do not need to be correlated with age. Dosages may be sustained release.

[0045] For tetracyclines, immunosuppressants, and statins, gradual dose escalation may be utilized. Dose escalation may be used to make the treatment more potent, to mitigate adverse side effects caused by the drug, or to allow side effects to appear later in the treatment, reducing patient discomfort. The described treatment methods may be modified by gradual dose escalation. Gradual dose escalation may occur over days or weeks, such as 25% of the total dose the first day or week, 50% the second, 75% the third, and 100% thereafter. Other gradual dose escalations may take the form of 50% dose for the first period, then 100% dose.

[0046] Other antibiotics that inhibit protein gene transcription, translation, and synthesis may be used. The tetracyclines used may be synthetic, semi-synthetic, or naturally derived. In addition, if more synthetic tetracyclines are invented in the future, they will likely be useful as well, since the basic structure of all tetracyclines is common. The following list of tetracyclines may be used: chlortetracycline, oxytetracycline, demeclocycline, lymecycline, meclocycline, methacycline, minocycline, rolitetracycline, glycecyclines, tigecycline, omadacycline, sarecycline, and eravacycline.

[0047] B cell function and amyloid beta and PrP sc Other immunosuppressants that block the synthesis of , such as cyclosporine, tacrolimus, and everolimus, may be used. Macrolides may also be used. Suitable macrolides include azithromycin, clarithromycin, erythromycin, fidaxomicin, and telithromycin. Other immunosuppressants, such as corticosteroids, Janus kinase inhibitors, calcineurin inhibitors, mTOR inhibitors, IMDH inhibitors, biologics, and monoclonal antibodies, may also be used.

[0048] For type III and APOE ε4 positive patients, statins may be used. Suitable statins include atovar, atorvastatin, cerivastatin, fluvastatin, lovastatin, mevastatin, pitavastatin, pravastatin, fenofibric acid, alirocumab, etravirine, cyproterone, posaconazole, ticagrelor, bezafibrate, coenzyme Q-10 colesevelam, rosuvastatin, simvastatin, simvastatin + ezetimibe, lovastatin + niacin extended release, atorvastatin + amlodipine, and simvastatin + niacin extended release.

[0049] Certain patients undergoing AD therapy may require a triple combination approach. Patients may require a therapy consisting of minocycline, sirolimus, and atorvastatin. Atorvastatin is also known as Lipitor®. The dosage of atorvastatin can range from 1 mg to 100 mg. Suitable dosages may include 50 mg minocycline, 1 mg sirolimus, and 10 mg atorvastatin. If atorvastatin is not used and another statin must be used, the other statin may range from 0.01 mg to 200 mg. For example, if cerivastatin is used, the suitable dosage will be much less, such as 0.8 mg cerivastatin, 50 mg minocycline, and 1 mg sirolimus.

[0050] Despite the failure of BACE inhibitors in treating AD, the amyloid protein deposition hypothesis remains valid. These studies failed because the inhibitors acted downstream of amyloid protein metabolism. The therapeutic methods described herein act at the level of DNA transcription into RNA and transport of RNA to mitochondria for protein synthesis by binding to the 30S and 50S subunits of RNA to inhibit amyloid protein synthesis. Furthermore, normal cellular catabolism removes already deposited amyloid beta protein.

[0051] Combination therapy for AD works. In physician-run trials, patients receiving triple therapy have had unexpected results. Treatment of several patients, including two identified herein as Patient A and Patient B, has demonstrated surprisingly good results with the appropriate doses and selection of combinations of antibiotics, immunosuppressants, and statins. The fact that this treatment worked at all, despite prior failures of others and abundant doubt in the field, would have been unexpected, and the fact that it worked so quickly was a welcome unexpected benefit.

[0052] Patient A is a 54-year-old APO E4 Patient A was a positive E3 / E4 male who had been suffering from AD for 5 years prior to treatment. Patient A had difficulty forming complete sentences and required the aid of a picture chart to communicate. Basic activities such as eating and going to the toilet required the patient to use hand gestures or point to pictures on the chart that illustrated the actions. Patient A was extremely frustrated and would sometimes become violent when his caregivers did not understand him. Patient A also had difficulty completing simple tasks. Patient A was unable to tie his own shoelaces and would often have repeated difficulties with door handles when trying to open doors. A word recall test was administered using an adaptation of the Alzheimer's Disease Assessment Scale prior to treatment. The patient was unable to recall or communicate any of the words that appeared on this test.

[0053] Patient A was treated with 2 mg sirolimus, 100 mg minocycline, and 20 mg atorvastatin once daily for 4 months. Patient A experienced recovery of speech, forming short phrases and occasionally completing entire sentences. Patient A's use of picture charts also increased, and Patient A became less irritable and abusive toward caregivers. Patient A's aphasia resolved, and he had improved short-term and long-term memory. A word recall test adapted from the Alzheimer's Disease Assessment Scale was administered, and the patient was able to remember up to five words.

[0054] Patient B is a 65-year-old APO patient with AD. E4Patient B is a positive male. Patient B had difficulty with daily tasks such as getting dressed, remembering meals, communicating with caregivers, and remembering family members. When buttoning his shirt, he would often forget to button it or the buttons would come out of place. He would sometimes forget clothes such as socks and underwear, and had difficulty putting on his shoes. The caregiver would have to prepare meals and remind Patient B to eat. Patient B also had difficulty remembering why the caregiver was in his house, and would repeatedly ask the caregiver about his identity. Patient B was able to remember and participate in conversations with his children, but would often ask who the children were in the middle of the conversation, as if they were strangers. Before treatment, a word recall test based on the Alzheimer's Disease Assessment Scale was administered. Patient B was able to recall one word each time he administered the test three times.

[0055] Patient B was treated with 2 mg sirolimus, 100 mg minocycline, and 20 mg atorvastatin once daily for two months. Patient B experienced improved short-term memory and improved long-term memory. Patient B no longer forgot to button his clothes and was perfectly able to dress himself. Patient B no longer forgot meals prepared by his caregiver. He asked less frequently who his children and caregivers were. After treatment, Patient B was able to recall up to four words in a word recall test based on the Alzheimer's Disease Assessment Scale.

[0056] Researchers have not been successful in treating AD because they have not understood the pathophysiology of the disease and how it works, and they have not realized that no single drug will be sufficient to treat the disease, which is why researchers have failed to use tetracyclines alone or statins alone to treat AD.

[0057] Conclusion Alzheimer's disease can be caused by a) intracerebral amyloidosis, b) systemic amyloidosis, or c) APO E4AD is a complex disease that presents several drug targets, including dyslipidemia in astrocytes and microglia in the CNS, d) brain neuroinflammation, and f) systemic inflammation caused by tumor necrosis factor α, interleukin 1b, and IL6. These targets require combination drug therapy. The simplest AD treatment is APO E4 Dual therapy and APO for negative (E2 / E3) AD E4 It can be simplified to a triple therapy for positive AD.

[0058] Throughout this specification, the illustrated embodiments and examples should be considered as examples, not limitations on the disclosed or claimed apparatus and procedures. Many of the examples illustrated herein include specific combinations of method acts or system elements, but it should be understood that those acts and those elements may be combined in other ways to achieve the same purpose. With respect to flow charts, additional and fewer steps may be taken, and steps as shown may be combined or further refined to achieve the methods described herein. Acts, elements, and features discussed only in connection with one embodiment are not intended to exclude a similar role in other embodiments.

[0059] As used herein, "plurality" means two or more. As used herein, a "set" of an item may include one or more of such items. As used herein, whether in the written description or the claims, the terms "comprising," "including," "carrying," "having," "containing," "involving," and the like, shall be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of," respectively, are closed or semi-closed transitional phrases with respect to the claims. The use of ordinal numbers such as "first," "second," "third," and the like in the claims to modify claim elements does not, in and of itself, imply a priority, precedence, or ordering of a claim element over another element, or a chronological order in which the acts of a method are performed, but are merely used as labels to distinguish a claim element having a name from another element having the same name (but using ordinal numbers) to distinguish between multiple claim elements. As used herein, "and / or" means that the listed items are alternatives, but that the alternatives include any combination of the listed items.

Claims

1. A pharmaceutical composition comprising an antibiotic or a pharma- ceutical acceptable salt thereof, an immunosuppressant or a pharma- ceutical acceptable salt thereof, a statin or a pharma- ceutical acceptable salt thereof, and one or more pharma- ceutical acceptable carriers, diluents, or excipients, wherein the antibiotic comprises 100 mg of minocycline, the immunosuppressant comprises 2 mg of sirolimus, and the statin comprises 20 mg of atorvastatin.

2. 2. The pharmaceutical composition of claim 1, wherein a patient is administered minocycline, sirolimus, and atorvastatin after being identified as having one of three types: Type I, Type II, or Type III, wherein Type I is characterized by Down's syndrome and occurs at about 30-40 years of age, and the genetic marker is trisomy 21, Type II is characterized by early onset Alzheimer's disease (EOAD) and occurs at about 50-60 years of age, and the genetic markers are PSEN1 and PSEN2 on chromosomes 14 and 1, respectively, and Type III is characterized by late onset Alzheimer's disease (LOAD) and occurs at about 65-90 years of age, and two-thirds of patients are positive for APOE ε4 located on chromosome 19.

3. Minocycline, sirolimus, and atorvastatin also cross the blood-brain barrier and inhibit PrP ｓｃ The pharmaceutical composition of claim 1 which targets a protein.

4. Prp ｓｃ The protein has more beta sheets than PrPc, and the PrP in blood ｓｃ The pharmaceutical composition of claim 3, wherein the amount of

5. PrP in blood after administration of the composition ｓｃ The pharmaceutical composition of claim 1 , wherein the protein is reduced.

6. 2. The pharmaceutical composition of claim 1, wherein minocycline, sirolimus, and atorvastatin inhibit the transcription of the APP gene and RNA transcription at the level of DNA transcription into transcribed RNA (tRNA), messenger RNA (cmRNA), RNA transport to mitochondria for protein synthesis and deposition in cerebral cortical neurons.

7. 2. The pharmaceutical composition of claim 1, wherein minocycline, sirolimus, and atorvastatin inhibit the synthesis of humoral antibodies and APP by the humoral immune system.

8. 2. The pharmaceutical composition of claim 1, wherein minocycline, sirolimus, and atorvastatin act at the level of DNA-to-RNA transcription and RNA transport to mitochondria for protein synthesis by binding to the 30S and 50S subunits of RNA and inhibiting amyloid protein synthesis.

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