WO2017070731A1

WO2017070731A1 - Compositions and methods for the treatment of alzheimer's disease

Info

Publication number: WO2017070731A1
Application number: PCT/AU2016/050353
Authority: WO
Inventors: Frederick Edward MAYELL
Original assignee: Innate Immunotherapeutics Limited
Priority date: 2015-10-28
Filing date: 2016-05-11
Publication date: 2017-05-04

Abstract

The present invention relates to compositions and methods for treating Alzheimer's disease. In particular, the present invention relates to the use of muramyl dipeptide crosslinked to form a microparticle for the treatment of Alzheimer's disease.

Description

Compositions and methods for the treatment of Alzheimer's disease Field of the Invention

[0001 ] The present application claims priority from Australian Patent Application No. 201590441 1 , filed on 28 October 2015.

[0002] The present invention relates to compositions and methods for the treatment of Alzheimer's disease. In particular, the present invention relates to the use of muramyl dipeptide crosslinked to form a microparticle for the treatment of Alzheimer's disease.

Background of the Invention

[0003] Any discussion of the prior art throughout the specification should in no way be considered as an admission that such prior art is widely known or forms part of common general knowledge in the field.

[0004] Alzheimer's disease is a neurodegenerative disorder of the central nervous system and the leading cause of a progressive dementia in the elderly population. The clinical symptoms of Alzheimer's disease include impairment of memory, cognition, temporal and local orientation, judgment and reasoning. Alzheimer's disease can also result in severe emotional disturbances.

[0005] Alzheimer's disease is the most common form of dementia, affecting up to 70% of all people with dementia. More than 5 million Americans may have Alzheimer's disease and it is one of leading causes of death in the United States.

[0006] Alzheimer's disease can be either sporadic or familial. Sporadic Alzheimer's disease can affect adults at any age, but usually occurs after age 65 and is the most common form of Alzheimer's disease. Familial Alzheimer's disease is a very rare genetic condition, caused by a mutation in one of several genes. The presence of mutated genes means that the person will eventually develop Alzheimer's disease, usually in their 40's or 50's.

[0007] Alzheimer's disease is characterized pathologically by the accumulation of extracellular β-amyloid plaques and intracellular neurofibrillary tangles in the brain. Diagnosis is made through the clinical assessment of the neurologic and neuropsychiatric signs and symptoms of Alzheimer's disease and the exclusion of other causes of dementia.

Alzheimer's disease is commonly classified into mild, moderate and severe stages by a brief cognitive screening examination, the Mini-Mental State Examination. Damage to the brain starts a decade or more before memory and other cognitive problems appear.

[0008] There is no cure for Alzheimer's disease and available treatments, such as tacrine, rivastigmine, galantamine and donepezil (acetylcholinesterase inhibitors) and memantine (an NMDA receptor antagonist) offer relatively small symptomatic benefit. No medication has been clearly shown to delay or halt the progression of the disease.

[0009] Accordingly, there is a need for new agents that are effective in the treatment of Alzheimer's disease.

[0010] It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

Summary of the Invention

[001 1 ] It has been surprisingly found that muramyl dipeptide crosslinked to form a microparticle (MDP microparticle) reduces the symptoms of Alzheimer's disease and reduces levels of biomarkers associated with Alzheimer's disease.

[0012] According to one aspect, the present invention provides a method for the treatment of Alzheimer's disease in a subject, comprising administering to the subject a composition comprising muramyl dipeptide crosslinked to form a microparticle.

[0013] According to another aspect, the present invention provides use of muramyl dipeptide crosslinked to form a microparticle for the manufacture of a medicament for the treatment of Alzheimer's disease in a subject.

[0014] According to another aspect, the present invention provides a composition comprising muramyl dipeptide crosslinked to form a microparticle for use in the treatment of Alzheimer's disease in a subject.

[0015] In one embodiment, the microparticle comprises DNA fragments. [0016] In another embodiment, the MDP microparticle is isolated from bacteria. [0017] In another embodiment, symptoms of Alzheimer's disease are reduced. [0018] In another embodiment, life expectancy of the subject is increased. [0019] In another embodiment, one or more biomarkers of Alzheimer's disease are reduced.

[0020] In another embodiment, the biomarkers are interleukin 6, interleukin 1 β, interleukin 8 and/or matrix metallopeptidase 9.

[0021 ] In another embodiment, the composition or medicament comprises one or more pharmaceutically-acceptable excipients, carriers, vehicles or diluents.

[0022] In another embodiment, the MDP microparticle is not encapsulated in a liposome.

[0023] In another embodiment, the composition or medicament is administered to the subject parenterally.

[0024] In another embodiment, the composition or medicament is administered to the subject intravenously.

[0025] In another embodiment, the composition or medicament is administered to the subject orally.

[0026] In another embodiment, the composition or medicament is administered to the subject at a dosage of about 1 μg to about 100μg.

[0027] In another embodiment, the composition or medicament is administered to the subject at a dosage of about 100μg to about 1000μg.

[0028] In another embodiment, the composition or medicament is administered to the subject at a dosage of about 100μg to about 700μg.

[0029] In another embodiment, the composition or medicament is administered to the subject at a dosage of about 300μg to about 700μg.

[0030] In another embodiment, the composition or medicament is administered to the subject at a dosage of about 500μg to about 700μg.

[0031 ] In another embodiment, the composition is administered to the subject at a dosage of about 1 , 25, 50, 75, 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 875, 900, 925, 950, 975 or 1000 μg. [0032] In another embodiment, the composition or medicament is administered to the subject once a day.

[0033] In another embodiment, the composition or medicament is administered to the subject once a week.

[0034] In another embodiment, the composition or medicament is administered to the subject once a fortnight.

[0035] In another embodiment, the composition or medicament is administered to the subject once a month.

[0036] The appropriate dosage of MDP microparticles, total amount administered and duration of administration can be easily determined by a medical practitioner based on guidance provided herein, the nature and severity of the Alzheimer's disease, and the response by the subject to the treatment. As an example, useful individual dosages may be selected from the range ^g to 1000μg of MDP microparticles, and may be administered once a day, once a week, once a fortnight or once month depending on the subject's condition, symptoms, tolerance and response to treatment. Doses in a higher range can also be used depending on the requirements, for examples doses in the range of 1000μg to 1500μg of MDP microparticles. Dosages at other frequencies may also be employed. An example of a suitable dosage regimen could be to start with an initial dose of 100 μg followed by escalated doses until appropriate beneficial therapeutic effects are observed in the subject, without significant side-effects. The dosage may be given as single bolus dose or infused over time, or given in divided doses. The total amount of MDP microparticles administered will depend on subject response and tolerance to treatment. The composition may be administered once a day, once a week, once a fortnight or once a month for a total period that depends on the subject's response.

[0037] MDP microparticle-containing compositions may be suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal, aerosol and/or parenteral administration. The compositions may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of MDP microparticle- containing composition which may be combined with a carrier material to produce a single dose may vary depending upon the subject being treated, and the particular mode of administration. [0038] The MDP microparticle-containing compositions may be administered alone or in combination with pharmaceutically acceptable excipients, carriers, vehicles or diluents, in either single or multiple doses. Suitable pharmaceutical acceptable excipients, carriers, vehicles and diluents include inert solid diluents or fillers, sterile aqueous solutions and various organic solvents. The compositions formed by combining the MDP-microparticle- containing compositions and the pharmaceutically acceptable excipients, carriers, vehicles or diluents are then readily administered in a variety of dosage forms such as tablets, powders, lozenges, syrups, injectable solutions and the like. These pharmaceutical compositions can, if desired, contain additional ingredients such as flavourings, binders, excipients and the like. Thus, for purposes of oral administration, tablets containing various excipients such as L- arginine, sodium citrate, calcium carbonate and calcium phosphate may be employed along with various disintegrates such as starch, alginic acid and certain complex silicates, together with binding agents such as polyvinylpyrrolidone, sucrose, gelatin and acacia. Additionally, lubricating agents such as magnesium stearate, sodium lauryl sulphate and talc are often useful for tabletting purposes. Solid composition of a similar type may also be employed as fillers in soft and hard filled gelatin capsules. Appropriate materials for this include lactose or milk sugar and high molecular weight polyethylene glycols. When aqueous suspensions or elixirs are desired for oral administration, the essential active ingredient therein may be combined with various sweetening or flavouring agents, colouring matter or dyes and, if desired, emulsifying or suspending agents, together with diluents such as water, ethanol, propylene glycol, glycerin and combinations thereof. The MDP microparticle-containing compositions may also comprise enterically coated dosage forms.

[0039] Suitable formulation protocols and suitable excipients, carriers, vehicles and diluents can be found in standard texts such as Remington: The Science and Practice of Pharmacy, 19^th Ed, 1995 (Mack Publishing Co. Pennsylvania, USA), British Pharmacopoeia, 2000, and the like.

[0040] It will be clear to those skilled in the art that MDP microparticle treatment may form a component of co-therapy in the treatment of Alzheimer's disease. Thus, MDP

microparticles may be administered in conjunction with other known therapies for Alzheimer's disease. Such known therapies include, but are not limited to, tacrine, rivastigmine, galanthamine, donepezil and memantine. Other co-therapy treatments include currently trialled monoclonal antibodies, immunotherapies and secretase inhibitors. In co-therapy treatment, MDP microparticles may be administered simultaneously or sequentially with other treatments. Definitions

[0041 ] In the context of the present invention, the terms "muramyl dipeptide crosslinked to form a microparticle" and "MDP microparticle" refer to a microparticle formed by crosslinked repeats of muramyl dipeptide (MDP), wherein the MDP repeats are crosslinked to each other. The MDP microparticle may also contain DNA fragments and/or other agents that stimulate and/or regulate the immune system.

[0042] In the context of the present invention, the words "comprise", "comprising" and the like are to be construed in their inclusive, as opposed to their exclusive, sense, that is in the sense of "including, but not limited to".

[0043] In the context of the present invention, the terms "treatment" or "treating" include preventing a disease, disorder or condition from occurring in a subject which may be predisposed to the disease, disorder or condition but has not yet been diagnosed as having it; inhibiting the disease, disorder or condition, e.g., impeding its progress; and relieving the disease, disorder, or condition, e.g., causing regression of the disease, disorder and/or condition or reducing the severity of the disease. Treating the disease or condition includes ameliorating at least one symptom of the particular disease or condition, even if the underlying pathophysiology is not affected.

[0044] In the context of the present invention, the phrase "therapeutically effective amount" refers to an amount of MDP microparticle that produces some desired effect at a reasonable benefit/risk ratio applicable to any medical treatment. In certain embodiments, the term refers to that amount necessary or sufficient to eliminate or reduce medical symptoms for a period of time. The effective amount may vary depending on such factors as the disease or condition being treated, the particular targeted constructs being administered, the size of the subject, or the severity of the disease or condition. One of ordinary skill in the art may empirically determine the effective amount of a particular composition without necessitating undue experimentation. In certain embodiments, compositions are formulated in a manner such that they will be delivered to a subject in a therapeutically effective amount, as part of a prophylactic or therapeutic treatment. The desired amount of the composition to be administered to a subject will depend on absorption, inactivation and excretion rates of the MDP microparticles, as well as the delivery rate of the MDP microparticle.

[0045] In the context of the present invention, the phrase "parenteral administration" refer to modes of administration other than enteral and topical administration, such as injections, and include without limitation intravenous, intramuscular, intrapleural, intravascular, intrapericardial, intraarterial, intrathecal, intracapsular, intraorbital, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.

[0046] In the context of the present invention, the phrase "pharmaceutically acceptable" is art-recognized. In certain embodiments, the term includes compositions, polymers and other materials and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of mammals, human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication,

commensurate with a reasonable benefit/risk ratio.

Brief Description of the Figures

[0047] Figure 1 : Schedule of MIS416 doses administered intravenously to Patient HM.

[0048] Figure 2: Improvement in symptoms in Patient HM after 5 months of MIS416 therapy.

[0049] Figure 3: Blood serum levels of Alzheimer's disease biomarkers (IL-1 p, IL-6, IL-8, MMP9) from Patient HM before MIS416 therapy and at time points during MIS416 treatment. All samples were stored at -80oC and analysed simultaneously.

Preferred Embodiment of the Invention

[0050] Although the invention has been described with reference to certain embodiments detailed herein, other embodiments can achieve the same or similar results. Variations and modifications of the invention will be obvious to those skilled in the art and the invention is intended to cover all such modifications and equivalents.

[0051 ] The present invention is further described by the following non-limiting examples. Examples

Example 1 : Preparation of MIS416

[0052] Propionibacterium acnes was grown to a mid-stationary growth phase and washed to remove contaminants of bacterial culture origin by employing techniques well known to those in the art. Hydrophobic components contained in the cell walls and cytoplasm were sequentially extracted by successive washes with increasing concentrations of

ethanol/isopropanol/water (10%:10%:80%, 25%:25%:50% and 40%:40%:20%) at elevated temperatures. The isopropanol was then removed with successive washes with decreasing concentrations (80%, 50%, 40% and 20%) of ethanol at elevated temperatures. The resultant microparticles (MIS416) were then suspended in 6M guanidine-HCI and then washed in water for irrigation. The concentration of MIS416 was measured by relating its absorbance at 565 nm to the absorbance of turbidity standards.

[0053] MIS416 contains extensively crosslinked MDP, amino-linked L-alanine-D- isoglutamine dipeptides and bacterial DNA fragments. The MIS416 generated by the present methods can have a broad range of sizes (for example, 0.01 to 30 microns) but the most common size range is from 1 to 7 microns. The preferred size is in the range of 0.5 to 3 microns.

[0054] MDP microparticles can be isolated from natural sources, as described above, or synthesized using well-known synthetic procedures (see, e.g., Liu et al., Bioorganic and Medicinal Chemistry Letters, 10 (12), 2000, pp. 1361 -1363(3); Schwartzman & Ribi, Prep Biochem. 1980; 10(3): 255-67; Ohya et al. Journal of Bioactive and Compatible Polymers, 1993; 8: 351 -364).

[0055] The concentration of MIS416 was adjusted to 0.2 mg/mL in sodium chloride for intravenous administration.

Example 2: MIS416 toxicology studies

[0056] It has been well established that free/soluble MDP has significant toxicity in vivo. Attempts to reduce MDP toxicity have employed procedures to delay release, such as MDP incorporation into liposomes or other related compounds, or modification of terminal groups. Chemical modification has resulted in marked reduction in activity, and designs which change delivery rate have been difficult to control.

[0057] In vivo toxicology studies for MIS416 were performed as summarized in Table 1 .

Study ID. Study Title Method Quantity of Outcome

MIS416

G6121 MIS416: MIS416 was administered 10,000, 15,000 Maximum tolerated

Acute as single escalating doses. 30,000 and dose (MTD) Toxicology Animals were monitored for 45,000 mcg/kg established as Study by IV toxic signs and mortality up body weight 15,000 mcg/kg route in to day 15 and subjected to body weight Swiss Albino detailed necropsy at

Mice terminal sacrifice on day 15 Study ID. Study Title Method Quantity of Outcome

MIS416

G6122 MIS416: MIS416 was administered 50, 200, 800 MTD established as

Acute as single escalating doses. and 3200 3,200 μg/kg body Toxicology Animals were monitored for mcg/kg body weight Study by IV toxic signs and mortality up weight

route in New to day 15 and subjected to (rabbits)

Zealand detailed necropsy at

White terminal sacrifice on day 15

Rabbits

G6123 MIS416: MIS416 doses selected 1 ,000 3,000 The no-observed- Four Week from acute toxicology study and 10,000 adverse-effect-level Repeated were administered twice mcg/kg body (NOAEL) was Dose weekly for 4 weeks. weight (mice) considered to be

Toxicology Animals were subjected to 1 ,000 mcg/kg (or a Study by IV detailed necropsy at total weekly dose of Route in terminal sacrifice. 2,000

Swiss Albino mcg/kg/week) Mice

G6124 MIS416: MIS416 doses selected 50, 500 and The NOAEL was

Four Week from acute toxicology study 5,000 mcg/kg considered to be 50 Repeated were administered twice body weight mcg/kg injected Dose weekly for 4 weeks. twice weekly (100

Toxicology Animals were subjected to mcg/kg/week) Study by IV detailed necropsy at

Route in New terminal sacrifice.

Zealand

White

Rabbits

1370-002 MIS416: A MIS416 was administered 20, 200, 1000 NOAEL in this

26-Week IV once weekly for 26 weeks. mcg/kg body study could be Toxicity Animals were subjected to weight considered as Study In detailed necropsy at being close to 20 Rabbits terminal sacrifice. A one mcg/kg for the

month recovery arm purpose of comprised 2 estimation of animals/sex/group. human safety

margins

Table 1 Summary of toxicology studies

[0058] The toxicity studies, conducted in mice and rabbits for up to 26 weeks duration, provide adequate safety margins to support long term clinical studies at dosage levels in patients up to 20 mcg/kg/week.

[0059] The toxicity studies show that MIS41 6 has significantly lower toxicity than free/soluble MDP. Example 3: Effect of MIS416 on Alzheimer's disease symptoms and prognosis

[0060] MIS416 was administered to patient HM, under the New Zealand "compassionate use" provisions according to the schedule set out in Figure 1 .

[0061 ] File notes were made based on observations by the husband and caregiver of patient HM, who is a retired doctor. The file notes are set out in Table 2.

3 July 2009

Caregiver reports that Patient HM's general condition is much the same as last

reported. Her general alertness continues. There has been no change in her memory or cognition. At this stage she has not taken up activities that she previously could not manage and there is nothing I can leave her to do by herself that she previously could not. She is happy and is always willing to help asking me what she can do to help.

13 August 2009

Caregiver reports that the improvement in Patient HM's general wellbeing has

continued. She is quite dramatically changed - more buoyant, happy, spontaneous, able to laugh at herself. This has been noticed by himself, friends and family.

Physically, Patient HM is in very good condition so there are no problems to report.

Previously she had become disinterested and lacking in energy. This has now

improved to the extent where she goes for long walks - in fact this week she managed a two hour walk with a friend on the same day as her last dose of MIS416. Caregiver is so impressed he is going to take her golfing (she used to be a good golfer but forgets where her ball is now) and see if he can reintroduce her to the game. She never has a reaction to the MIS416 - no chills or fever or headaches. The first few times she had muscle aches about 12 hours after injection but this no longer occurs.

Caregiver has expressed his satisfaction at her improvement although there is no change in her ability to do things for herself or in her intellectual capabilities. She cannot read, cook a meal on her own, cannot even use the telephone book. He is going to stop her driving as she is losing the ability to make quick decisions.

10 September 2009

Patient HM has now been receiving MIS416 on a weekly basis since 18 March 2009. Of this time, she has been on a 500 microgram dose for the last 12 weeks.

Caregiver reports that Patient HM continues to be more connected and several friends who have not seen her for 6 months or so have remarked on her marked increased buoyancy and alertness. Patient HM's mood level is very good and she feels very well and happy in herself, although she aware of her shortcomings as far as memory and cognition which has its difficulties. She continues to walk daily-up to an hour. She has not played golf as yet due to weather and time but caregiver intends to take her out in the next few weeks.

15 October 2009

Caregiver reports that Patient HM still continues to be very bright and happy in herself and continues to exercise regularly. She was recently at a social function where some of her old bridge friends who had not seen her for a while where there. They all remarked on how good she was compared when they last saw her.

Caregiver does do not think that there has been any change in her memory or cognition but, from his point of view the treatment has made life much easier - caregiver and Patient HM have lots of fun and try to laugh at her mistakes but obviously there are difficulties.

26 May 2010

Caregiver reports that the 2 doses of 700 microgram provoked a mild reaction. Patient HM is very happy. While caregiver cannot say her memory or cognition has improved, he feels that MIS416 has had a beneficial effect and is well worth continuing."

30 August 2010

Patient HM was diagnosed with Alzheimer's disease 8 years ago but had symptoms for 2 years prior. Her Alzheimer's disease has progressed more slowly than is usual. Many are in care in 2-3 years.

Caregiver reports that Patient HM saw a psychiatrist yesterday who assessed her and discussed MIS416 which she has now been on since 18th March 2009. She had weekly infusions gradually building up to 500mcg on the 17th June on which she stayed until a few months ago (31 /3/10 to 650 fortnightly and 28/04/10 to 700 micrograms three-weekly) when the dose was increased slowly at 3 weekly intervals to 700mcg. At this dosage Patient HM had her first ever chills and had paracetamol. She has continued with this dose and takes 2 paracetamol prior which has alleviated the aches that she was getting.

Patient HM's disease is progressive and caregiver reports that it has progressed since being on MMS416 but cannot say if the rate of progression has been reduced by the drug. The psychiatrist pointed out that if she had not been on the drug progression could have been much faster. However, there is no way to tell.

Care giver reports that, even if there is doubt or inconclusive evidence to support the reduction in progression, the very positive aspect of Patient HM's treatment is that she has been very happy and buoyant as to her mood level which is not the norm in most Alzheimer's disease people. Whilst she cannot do the things she used to do (reading writing playing sport, looking after grandchildren) she now knits, paints with a group and walks regularly and for this reason alone the caregiver wishes to continue with the infusion.

7 March 2011

Caregiver reports that Patient HM's Alzheimer's disease has progressed at about the same slow rate since the last report (6 months ago). Most noticeable is the now almost instant memory loss and the inability to perform simple tasks, for example operating the TV remote, setting the table, putting things away in the right place, making telephone calls. Her cognition is very limited. On the positive side she is happy within herself - placid and non argumentative. She is aware of how her Alzheimer's disease has affected both their lives and wishes she could do the things she has done in the past. Patient HM has always been a good sleeper (9-10 hours) and this pattern has continued. She continues to enjoy her weekly art group and has just started weekly Tai Chi classes at a nearby retirement village. She listens to music (difficulty in changing the discs) and walks regularly. She loves company although her contribution to conversation is virtually nil.

Patient HM tolerates 700mcg MIS416 now 4 weekly extremely well. She has two paracetamol prior to her injection. There have been no noticeable changes to extending the dose to four weekly or anything to suggest more frequent dosage would be better. Again there is no way to tell if MIS 416 has slowed the process down but she has deteriorated more slowly than one would expect from the usual Alzheimer's disease patient. She continues to take donepezil 10mgm daily which she has been taking for 9 years. Physically Patient HM has no problems. Caregiver is keen for her to continue her treatment.

8 June 2011

Caregiver reports that, apart from her Alzheimer's disease, Patient HM is physically very well and has had no deterioration. She exercises regularly and is happy. Carer thinks that MIS416 is contributing to her well being.

2 December 2011

Caregiver reports that Patient HM is much the same but with almost immediate memory loss. She is very happy in herself although she is aware of the things she can't do that she used to which at times makes her sad. She has now had Alzheimer's Disease for 10 years. Friend remarked to carer that he thought the treatment was helping with the progression. Patient HM has not had any illnesses, including winter ailments, for 3years. People with Alzheimer's disease are far more susceptible. Maybe

immunological effect of MIS416 is helping in this respect.

3 July 2012

Patient HM received her monthly injection (at 700 micrograms) of MIS416 for her Alzheimer's Disease. This is week 172 of treatment and her 85th dose.

Caregiver reports that Patient HM is still slowly deteriorating but is very happy. She is able to perform all her daily needs and recognize and name sometimes with prompting family and friends. She is still walking and has a route near home which she can do by herself. She is unable to play golf. Patient HM is more confused than what she was a year ago. Her mother (aged 94) died 3 weeks ago which has not helped but all in all she has handled it well.

5 June 2013

Patient has just had her 220th injection of MIS, still continuing at 700 micrograms per month with Paracetamol prior. She never shows any kind of reaction to the treatment.

Caregiver reports that Patient HM's slow deterioration continues and that he now have to help her with regulating her shower and help with her dressing. She is very limited as to what she can do. Now into the 13th year and the caregiver likes to think the MIS414 has slowed the progress but who knows.

31 March 2014

Caregiver reports that he has not been administering MIS416 to Patient HM for some six months now as her Alzheimer's Disease has now progressed to the point where he feels the treatment is no longer of benefit.

Table 2: File notes for treatment of Patient HM [0062] A comparison of Patient HM's symptoms before treatment and after 5 month's treatment is shown in Figure 2.

Example 4: Effect of MIS416 on Alzheimer's disease biomarkers

[0063] Peripheral blood serum from Patient HM was collected at baseline (pre-treatment) and at several time points during MIS416 therapy and stored at -80°C until analysis.

[0064] Biomarkers associated with Alzheimer's disease, specifically interleukin 6 (IL-6), interleukin-1 beta (IL-1 β), interleukin 8 (IL-8) and matrix metallopeptidase 9 (MMP-9) were quantified using flow cytometry bead based methodology on serial serum samples, which were assayed simultaneously.

[0065] A reduction was observed for IL-6, IL-1 p, IL-8 and MMP-9 (Figure 2),

demonstrating that MIS416 therapy was associated with a clear reduction in Alzheimer's disease biomarkers which were highly elevated in pre-treatment sample. This reduction was sustained for period of assessment (1 13 weeks of MIS416 treatment).

Claims

1 . A method for treatment of Alzheimer's disease in a subject, comprising administering to the subject a composition comprising muramyl dipeptide crosslinked to form a

microparticle.

2. The method according to claim 1 , wherein the microparticle comprises DNA fragments.

3. The method according to claim 1 , wherein symptoms of Alzheimer's disease are reduced.

4. The method according to claim 1 , wherein life expectancy of the subject is increased.

5. The method according to claim 1 , wherein one or more biomarkers of Alzheimer's disease are reduced.

6. The method according to claim 5, wherein the biomarkers are selected from interleukin 6, interleukin 1 β, interleukin 8 and matrix metallopeptidase 9.

7. The method according to claim 1 , wherein the composition is administered to the subject parenterally.

8. The method according to claim 1 , wherein the composition is administered to the subject intravenously.

9. The method according to claim 1 , wherein the composition is administered to the subject orally.

10. The method according to claim 1 , wherein the composition is administered to the subject at a dosage of about 1 μg to about 10C^g.

1 1 . The method according to claim 1 , wherein the composition is administered to the subject at a dosage of about 10C^g to about 10OC^g.

12. The method according to claim 1 , wherein the composition is administered to the subject at a dosage of about 50C^g to about 70C^g.

13. The method according to claim 1 , wherein the composition comprises one or more pharmaceutically-acceptable excipients, carriers, vehicles or diluents.

14. The method according to claim 1 , wherein the composition is administered to the subject once a day.

15. The method according to claim 1 , wherein the muramyl composition is administered to the subject once a week.

16. The method according to claim 1 , wherein the muramyl composition is administered to the subject once a fortnight.

17. The method according to claim 1 , wherein the muramyl composition is administered to the subject once a month.

18. Use of muramyl dipeptide crosslinked to form a microparticle for the manufacture of a medicament for the treatment of Alzheimer's disease in a subject.

19. A composition comprising muramyl dipeptide crosslinked to form a microparticle for use in the treatment of Alzheimer's disease in a subject.