IN AUTOIMMUNE DISEASE TREATMENT

INVENTION DESCRIPTION

1. FIELD OF APPLICATION

The invention relates to combination of pro-opiomelanocortin endogenous peptides ACTH 1-13 adrenocorticotropic hormone and metenkefalin, in treating autoimmune diseases, in the field of peptide and protein application, in the pharmaceutical industry and medicine.

In the International Patent Classification, it is classified as Section A - Human Necessities, Class A61 - Medical Science, Subclasses A61 K - Preparations for Medical Purposes, and A61 P - Therapeutic Activity of Medical Preparations, and also as C - Chemistry, C07K - Peptides.

2. TECHNICAL PROBLEM

The technical problem is the lack of knowledge on efficient and clearly defined doses, adequate to the disease and disabledness level and degree, related to particular diseases, as well as the way of defining the dose in the technological sense.

So far it was not clear whether technologically both these peptides are to be applied to the patients separately or together, in a single mixture, as a single dose.

Furthermore, it was necessary more clearly to define the route and manner of application of the defined doses to the patients. A correct and clear dose is to solve the following problems:

- corticosteroid substitution therapy in the disease relapse, because of the corticosteroid side effects, with determining the efficient POMC peptides dose and dose ranges;

- establishing the dose ranges, adequate to the disease degree, aimed to activation of the defence growth mechanisms and potentials, in treating of relapsing remitting multiple sclerosis RRMS, secondary progressive multiple sclerosis SPMS, and primary progressive multiple sclerosis PPMS, with clear determination of dosing of both POMC peptides in a single dose;

- determining the size of the depot dose in a POMC peptides therapy in treating RRMS, SPMS and PPMS, that is both safe and efficient;

- determining the most efficient route of application of POMC peptides in treating RRMS, SPMS and PPMS.

3. STATE OF THE ART

In the scientific literature and the medical research practice, often accepted is the principle of the effects of a substance or medicine being proven when these are proven on an animal model.

However, the right clarifications for particular industrial application, that is, application on patients can be noticed and the problem can be solved only by actual research on humans.

Adrenocorticotropic hormone ACTH 1-13 and metenkefalin are two peptides from the region of the pro-opiomelanocortin hormone POMC, that have mutually additive effect that has been proven on mouse model of ulcerous colitis and patented under the name COMBINATION OF TWO BIOACTIVE REGIONS OF PRO-OPIOMELANOCORTIN HORMONE, EP 1 435 999.

While researching the said patent, the inventors made knew particular knowledge that make new inventive steps in dosing, aimed to activating the treatment mechanisms and providing new possibilities and knowledge for industrial application, which steps are to be protected as a new patent

The certified patent solution has established the efficient dose of 1 mg of alpha MSH, that is analogous to ACTH 1-13 + 10 mg of metenkefalin, these making an 1 1 mg POMC peptide combination dose.

Since here as well the efficient dose, required by the patent, has been proven on animal model only, only researching the patent in human studies has produced efficient doses that are acceptable and precise, and as such positive proves related to application in human medicine. Noted are the levels of needs to increase quantities of each of the peptides in the combination that will increase the total dose of the POMC peptides, in order to achieve the effect and activation of particular therapeutic potentials for an efficient treatment.

At the same time, animal models make basis for the assumed therapeutic application that may include a larger number of indications in the same pathogenesis, although there are no specific proofs of this.

The above mentioned patent states no specific disease to which it relates. Now, however, based on the knowledge acquired in the research, we may state specific diseases to which the POMC peptides combination relates.

4. ESSENCE OF THE INVENTION

The essence of the invention is combination of pro-opiomelanocortin endogenous peptides: adrenocorticotropic hormone ACTH 1-13 and metenkefalin, in autoimmune disease treatment, where dose of the POMC peptides combination in human application is increased by at least doubling quantity of the peptide ACTH 1-13, so that the 12 mg starting dose containing 2 mg of ACTH 1-13 + 10 mg of metenkefalin, instead of the 1 1 mg dose as stated in the above mentioned patent that related to 1 mg of alpha MSH, which is another name for ACTH 1-13, + 10 mg of metenkefalin.

Hence, this invention at least doubles one of the peptides in the combination to achieve the effect, so that a dose contains from 2 mg to 10 mg of ACTH 1-13 and from 10 mg to 50 mg of metenkefalin, that may provide:

- success in substitution of corticosteroids in disease relapse, in the suggested dosage raster;

- growth and increase of dose of both POMC peptides, in accordance with a patient's disease degree and gravity, in order to activate the interleukin IL10 mechanisms and growth potentials, the growth of which inhibits, prevents, chromosome aberrations, this helping tissue growth and regeneration, inhibits pro-inflammatory cytokines growth and, thereby, slows inflammation down; furthermore, it is of particular importance in treating autoimmune diseases that the IL10 enables modulation of the immune response from the Th1 , which is a wrong one, into the new Th2;

- dosage for the secondary progressive multiple sclerosis, SPMS, based on the proves from the pilot study of the people suffering from the SPMS. 5. BRIEF DESCRIPTION OF ILLUSTRATIONS

Figure 1 Cortisol level one hour after the corticosteroid pulse therapy is increased over four times relative to the physiological values.

Figure 2 Cortisol level in POMC peptides (ACTH 1-13 and metenkefalin) therapy has remained within the physiological value limits.

Figure 3 Significant rise of Cortisol in patients treated with corticosteroids relative to patients treated wit the ACTH 1-13 and metenkefalin combination, meaning also a significant increase of side effects.

Figure 4 Decrease of chromosome aberrations.

Figure 5 Significant decrease of chromosome aberrations with the ACTH 1-13 peptides

+ metenkefalin combination proves this to be an efficient therapy aimed to slowing down the demyelisation processes in patients with multiple sclerosis, as compared with corticosteroid therapy, with a quicker recovery.

Figure 6 EDSS (Expanded Disability Status Scale) disability degree is significantly lesser in the group of patients with the secondary progressive multiple sclerosis SPMS that was treated with the POMC ACTH 1-13 peptides + metenkefalin combination in the doses prescribed for the SPMS.

Figure 7 Significantly lesser EDSS disability degree in patients with the secondary progressive multiple sclerosis SPMS who were administered larger doses of the ACTH 1-13 peptides + metenkefalin combination.

Figure 8 Increase of the IL10 in patients with multiple sclerosis before treatment with the ACTH 1 -13 peptides + metenkefalin combination (IL10A) and after the treatment (IL10B), proving modulation of the immune response from Th1 into Th2 and inhibition of the pro-inflammatory cytokines.

Figure 9 Advantage of nasal application of the POMC ACTH 1-13 peptides and metenkefalin: the shortest route to the central nervous system CNS is the nasal one, which is advantageous in neurological diseases for the following reasons:

- faster acting in the CNS due to the shorter route to the CNS

- possibility of greater concentration of the medicine in the CNS. All the results shown in the above figures are generated in the comparative study of administering corticosteroids to patients with multiple sclerosis in the course of the disease relapse.

6. DETAILED DESCRIPTION OF THE INVENTION a) Combination of proopiomelanocortin endogenous peptides

Combination of pro-opiomelanocortin POMC endogenous peptides is a combination of ACTH 1-13 adrenocorticotropic hormone and metenkefalin.

The said POMC combination is applied as the therapy in disease relapse, that is, in acute exacerbation seizures, with a range of doses containing active substances in the peptide form: ACTH 1 -13 from 2 mg to 10 mg and metenkefalin from 10 mg to 50 mg. The said combination is applied in a single lyophilisation cake, as a single dose of 12 mg to 60 mg, for preparation of solution diluted with physiological solution 0.9% NaCI or distilled water, for 2 to 30 consecutive days, or until the patient is out of the disease relapse, where, depending on the disease degree, doses of 2 mg of ACTH 1 -13 and 10 mg of metenkefalin are increased within the protected therapeutic range, as required. The said combination is prepared for subcutaneous s.c, intramuscular, intravenous or oral application, as tablets, spray, lingualettes, drops, spray, rectal suppositories, cream or gel, as eye drops, or as nasal drops and spray.

The invention makes substitution of the corticosteroid therapy in the disease relapse and at determining the dose level to substitute corticosteroids in relapse, but with no corticosteroid side effects. Subsequent researches have proven success in substituting the corticosteroid therapy with the POMC peptides combination, in a dose that is greater then the presently known and efficient dose of 1 mg of ACTH 1-13 and 10 mg of metenkefalin, that produced a dose of 11 mg of combination of the two peptides.

There has been noticed requirement of double participation of ACTH 1-13, in order to increase the effects, which has produced a result that can successfully stop the relapse. Results of this comparative study are presented in the Figures 1 , 2 and 3.

It is evident that the Cortisol level increased by over four times in patients treated with the corticosteroids pulse therapy, Figure 1 , whereas in the patients treated with the ACTH 1- 13 and metenkefalin combination the Cortisol level remained within the physiological value limits, Figure 2.

The side effect level correlates with the Cortisol rise, which proves the effect of the ACTH 1-13 and metenkefalin combination effect in treating disease relapses with 12 mg doses, that is, 2 mg of ACTH 1-13 and 10 mg of metenkefalin, free of the corticosteroid side effects. This is evident in results of the comparative study of the corticosteroid therapy substitution, Figure 3. b) Dosing and dose sizes for particular indications

For relapsing remitting multiple sclerosis RRMS, secondary progressive multiple sclerosis SPMS, primary progressive multiple sclerosis PPMS, and asthma, of importance is the size of the dose, because the effect has been noticed to be increased by dose concentration, achieved by repeated applications in grave, acute conditions such as disease relapses - seizures.

Therefore solving the problem is oriented to dosing that is increased aliquot to the disease gravity.

In the trials of consequential dynamics of application in the disease relapse - seizure, noticed is the effect of cumulative acting of the POMC peptides combination, which justifies the need to increase the dose adequate to the disease degree and the patient's condition, provided it is kept within therapeutic range of the applied doses, as these are established in animal toxicological studies.

Therefore, established are the doses increased gradually adequate to the disease gravity and the subsequently acquired information.

Studies have established that the ACTH 1-13 and metenkefalin combination, when adequately dosed, decreases chromosome aberrations in patients with various degrees of disease, vital function disability and decreasing the life quality in general. This is a certain way to slowing demyelisation processes down and to remyelinasion of the lesions created in inflammatory process in the patients with multiple sclerosis. Decrease of chromosome aberrations in the comparative study of safety and efficiency of the POMC peptides, ACTH 1-13 and metenkefalin, in relapse compared to corticosteroid therapy, in patients with multiple sclerosis, is presented in the Figures 4 and 5. The following doses are established:

- 2 mg of ACTH 1-13 + 10 mg of metenkefalin, applied in a single 12 mg lyophilisation cake to treat relapsing remitting and secondary progressive multiple sclerosis and other autoimmune diseases, Alzheimer disease, Parkinson disease, epilepsy, asthma and allergic diseases. The proven efficiency in treating the secondary progressive multiple sclerosis, SPMS, has become evident in the pilot study of patients with this disease, Figures 6 and 7.

- 3 mg of ACTH 1-13 + 15 mg of metenkefalin, applied in a single 18 mg lyophilisation cake in cases of medium grave diseases and dysfunctions, in treating primary progressive and secondary progressive multiple sclerosis and other autoimmune diseases, Alzheimer disease, Parkinson disease, epilepsy, asthma, chronic obstructive pulmonary disease COPD and allergic diseases.

- 4 mg of ACTH 1-13 + 20 mg of metenkefalin, applied in a single 24 mg lyophilisation cake for grave diseases with high degrees of disease and dysfunction, in treating primary progressive multiple sclerosis and other autoimmune diseases, Alzheimer disease, Parkinson disease, epilepsy, asthma, chronic obstructive pulmonary disease COPD and allergic diseases.

All the above doses are prepared with both peptides in a single lyophilisation cake, to be applied subcutaneously s.c, intramuscularly, intravenously or orally, as tablets, spray, lingualettes, drops, spray, rectal suppositories, cream or gel, as eye drops, or as nasal drops and spray. c) Determining the depot dose for specific patients with increased disability degree in certain chronic inflammatory, autoimmune and allergic diseases

In the comparative study of effects of the POMC peptides, that is, ACTH 1-13 and metenkefalin combination, in disease relapse, relative to the corticosteroid pulse therapy, noticed is a significant increase of the IL10 interleukin in the patients treated with the POMC peptides combination.

Having in mind the overall effect of IL10, that is significant in modulation of incorrect immune response, especially in autoimmune diseases, because of its significance in inhibiting the effects of anti-inflammatory cytokines and inhibiting the apoptosis process, predicted is a significant effect of the depot dose. Evidences of the IL10 increase are shown in the Figure 8.

In patients with high disability and disease degrees, and aimed to decreasing the drug application dynamics, with possibility of applying depot dose also with any other patient requiring the ACTH 1-13 + metenkefalin combination therapy, but within the toxicoiogically safe range, the depot dose is determined within the following limits of contents of the POMC active substance: from 4 mg to 10 mg of ACTH 1-13 + 20 mg to 50 mg of metenkefalin in a single lyophilisation cake, as a single 24 mg to 60 mg dose, to be applied subcutaneously s.c, intramuscularly, intravenously or orally, as tablets, spray, lingualettes, drops, spray, rectal suppositories, cream or gel, as eye drops, or as nasal drops and spray, the said doses containing:

- 4 mg of ACTH 1-13 + 20 mg of metenkefalin, applied in a single 24 mg lyophilisation cake;

- 6 mg of ACTH 1-13 + 30 mg of metenkefalin, applied in a single 36 mg lyophilisation cake;

- 10 mg of ACTH 1-13 + 50 mg of metenkefalin, applied in a single 60 mg lyophilisation cake.

Depot dose is applied in cases of grave conditions of patients with secondary progressive and primary progressive multiple sclerosis, asthma, epilepsy and other chronic inflammatory and grave allergic diseases.

The most effective route of application of the doses stated in the above points a), b) and c) to overcome the blood brain barrier BBB by the POMC peptides combination (ACTH 1-13 + metenkefalin), aimed to increasing the effects and decreasing the side effects

POMC is located on the 2p23.3. chromosome. POMC appears on anterior and intermediary hypophysis lobes, and codes the 285 amino-acid long polypeptide hormonal precursor, which means that all POMC parts, peptides, are too products of the very central nervous system, CNS. This is why they are referred to as endogenous.

The POMC peptides, ACTH 1-13 and metenkefalin, combination is ideal for nasal application, especially in treating neurological diseases, related to problems in the CNS, but also some other autoimmune and inflammatory diseases, and this for the following reasons:

- nasal route lets amino acids through unless they are large molecules, and POMC peptides, ACTH 1-13 and metenkefalin, are small molecules, that additionally enables easier passage by the intranasal route and passing the BBB;

- the ACTH 1-13 and metenkefalin combination, when joined in a single lyophilisation cake, is highly and quickly soluble, that additionally enables easier passage by the intranasal route and passing the BBB;

- the ACTH 1-13 and metenkefalin combination is endogenously designed, the organism does not feel it as a foreign body and will show no resistance and side effects while passing the nasal route, and therefore also the BBB;

- the ACTH 1-13 and metenkefalin combination is produced by the very CNS, that is, these peptides are synthesized in the very CNS (the hypophysis) within the POMC molecule, which means that the very CNS recognises them as its own peptides and does not create a barrier preventing their entering into the CNS. The entire recognition mechanism is performed by the key-keyhole system, so that passing of the ACTH 1-13 and metenkefalin combination into the CNS, that is, passing the BBB, is perfectly certain and without resistance, which again means without marked side effects in application;

- nasal application is the shortest and the fastest route to the CNS, as shown in the Figure 9;

- nasal application enables application of the drug, instead of by injection that most patients are afraid of and whose puncture is another side effect on the skin, as nasal spray, as a new way of application with a nasal application set, by atomising the active substances, that is, the POMC peptides, and their application as a spray.

The invention is applied for application of the medicine by the intranasal route, that is, by nasal application, as the direct and shortest route to the CNS, to overcome the blood brain barrier BBB, thereby as a substitution for injection, designed as a complete set with all the elements necessary for direct application to the patient, regardless of whether the patient applies the medicine by himself or is aided in this by another person. The dose that contains active substances of peptides is applied nasally, in the form of the POMC peptides, ACTH 1-13 + metenkefalin, combination, that successfully passes through the BBB when applied nasally, with a nasal application set that contains:

- separate doses in lyophilisat, with the diluter - physiological solution or distilled water,

- injection needles for diluting the lyophilisat,

- separate 2-5 ml syringes for taking the diluted lyophilisat from the container,

- atomisers that are to be fitted on the syringe after diluting the lyophilisat and taking the needle off the syringe, so that the diluted active substance from the syringe is applied nasally atomised.

ABBREVIATIONS IN THE INVENTION DESCRIPTION HAVE THE FOLLOWING MEANINGS:

POMC - pro-opiomelanocortin

ACTH 1-13 - adrenocorticotropic hormone 1-13

Alpha MSH - alpha melanotropin (analogue to ACTH 1-13)

Th1 and Th2 - therapeutic response

MS - multiple sclerosis

RRMS - relapsing remitting multiple sclerosis

SPMS - secondary progressive multiple sclerosis

PPMS - primary progressive multiple sclerosis

CNS - central nervous system

IL10 - interleukin 10

NaCI - sodium chloride

COPD - chronic obstructive pulmonary disease

s.c. - subcutaneously

BBB - blood brain barrier

7. INVENTION APPLICATION

The invention is applied in treating of patients under seizure (relapse) of chronic inflammatory and allergic, and especially autoimmune, diseases, when their only option is corticosteroid therapy, which is a hard therapy with numerous serious side effects, that also crates permanent damages in the patient's other organs and life functions, the invention being a substitute therapy free of side effects.

The invention is specifically applied for gradual dosing in accordance with the disease degree that is being treated, and to decrease discomforts of the patients with grave multiple sclerosis conditions, in the forms of advanced phases of relapsing remitting, secondary progressive and primary progressive multiple sclerosis, as well as generally in chronic inflammatory and allergic diseases.

The invention is also applied for exceptionally grave conditions of patients depending on receiving assistance from other persons, mostly immobile patients, in order to decrease the number of applications by a depot dose, in accordance with the degree of disability. The invention is also applied to apply the drug by the intranasal route, that is, by nasal application, as the direct and shortest route to the CNS. The dose containing the peptide active substance is applied by spray and a nasal set.