LT96079A - Use of corticotropin-releasing factor for the manufacture of a medicaments - Google Patents

Description

The present invention relates mainly to the use of corticotropin releasing factor (CRF) or analogues thereof for reducing swelling when episodic exacerbations occur in chronic inflammation. Among those suffering from human illnesses are chronic inflammatory arthritis, which include rheumatoid arthritis, Reiter's syndrome, ankylosing spondylitis, psoriatic arthritis and inflammatory bowel disease. Rheumatoid Arthritis alone In the United States, there are about 33,000 causes of deaths each year. So far, this disease is incurable and the cause is not clear. With arthritis, "outbreaks" or episodic exacerbations occur from time to time, extending from week to month.

Steinman ("Autoimmune Disease", Scientific American, pp.107-114, September, 1993) discusses a number of possible novel treatment methods for autoimmune diseases such as rheumatoid arthritis. One of them suggests the creation of mock-ups that would fit into a HLA (human lymphocyte antigen) crack when trying to eliminate T cells. According to another approach, a monoclonal antibody that binds to a tumor necrosis factor should be considered to increase joint motility and reduce rigidity associated with rheumatoid arthritis. However, to date, it has not been confirmed that these possible new treatments will be therapeutically effective.

Animal models were developed and refined using arthritic rat lines to help find treatments. In addition to other possible abnormalities, Liuiso's arthritic rats have a deficient hypothalamic CRF response to various stimuli, and therefore researchers may experimentally induce inflammatory arthritis and some other autoimmune diseases. In the Piebald-Viral-Glaxo (“PVG”) rat model, unlike the Lice line rats, the animals do not appear to have a hypothalamic-pituitary-adrenal (“HPA”) axis defect but are still susceptible to arthritis. U.S. Pat. No. 5,563,303 (issued April 9, 1991, inventors Sternberg et al.) Describes the use of a Louis rat animal model in which experimental animals have arthritis as a response to group A streptococcal cell wall fragments by modeling human rheumatoid arthritis. . Sternberg et al. shows how this model can be used to determine the susceptibility of mammals to inflammatory diseases. Sternberg et al. describes the measurement of CRH (corticotropin releasing hormone) after administration of 2 different agents. CRH is another name for a corticotropin releasing factor (hereinafter referred to as CRF), which is a 41 amino acid neuropeptide located in the brain and peripheral nerve endings, which stimulates the release of ACTH from the pituitary to the brain and from peripheral states such as nerve endings and leukocytes. . Sternberg et al. also describes injections of CRF and subsequent measurements of ACTH and corticosterone.

Antibodies directed against CRF have been described as partially eliminating the inflammatory response, and this has been suggested by various researchers to suggest that CRF is a proinflammatory agent. [King and others. "Autocrine or paracrine inflammatory corticotropin releasing hormone in vivo", Science, 254.421-423 (1991). U.S. Patent 4,489,163 to Rivier et al., Published December 18, 1984, describes CRF and its analogs in rats. Human CRF has the same sequence as CRF in rats. There are many CRF analogues known in the art. U.S. Patent No. 4,415,558 (inventors Vale, Jr., et al., Issued November 15, 1983) describes the synthesis, analogs and oCRF of sheep CRF from extracts of the ram hypothalamus. It has been found that synthetic oCRF reduces blood pressure.

A very similar peptide - souvazine - is described in Reguliatory Peptides, 2, pp.1-13 (1981). Souvenzine is a 40-amino acid peptide, and its biological activity has been reported to reduce blood pressure in mammals and stimulate ACTH and β-endorphin secretion. U.S. Pat. No. 4,528,189 (inventors Leder et al., Published July 9, 1985) and U.S. Pat. No. 4,553,354 (inventors Leder et al., Published August 6, 1985) describe peptides similar to rat and rat. sheep CRF, and analogues thereof, and it was found that this white suction and carotid urotensin stimulates ACTH and reduces blood pressure, respectively. The CRF-related peptide, the white suction urotensin, has the same amino acid sequence as carotene urotensin except that the 24-position amino acid is isoleucine and the 27th is glutamic acid.

Ling et al., BBRC, 122 pp.1218-1224 (1984) describes a goat CRF structure that is the same as a sheep CRF. Esch et al., BBRC, 122, pp.899-905 (1984) describe a structure of a bull CRF that differs from a sheep and goat CRF with only one amino acid (position 33 includes asparagine rather than serine as goat and sheep). The pig CRF isolated and characterized by Patthy et al., Proc. Natl. Acad. Sci., 82, pp.8762-8766 (1985). The pig CRF has the same 3

the amino acid sequence (residues 1-39) as in rat / human CRF and differs from them only in the 40 and 41 positions. At position 40, either asparagine or isoleucine can be present, and the 41-residue is phenylalaninamide. U.S. Pat. No. 4,801,612 (inventor Wei, published January 31, 1989) describes the use of a chicotropin-releasing factor or analogues thereof for administration to a patient, an inflammatory response (acute) in the skin or mucosal membranes, and U.S. Pat. On August 11, 1992, inventor Wei) describes the use of CRF (or salts thereof, or analogues thereof) for treating patients with brain injury or disease, nervous system or musculoskeletal disorders, whose factor is edema.

Administration of corticotropin releasing factor (CRF) to patients with one of the chronic inflammatory arthritis, such as rheumatoid arthritis, reduces episodic exacerbations of inflammatory disease. The CRF doses of the present invention provide anti-inflammatory results, depending on the dose, even in the case of a chronic disease. Thus, in the practice of the invention, it is proposed to use CRF or its analogues for the treatment of such incurable diseases which do not give up to current therapy.

Treatment should be carried out under the supervision of health professionals, usually therapists, and may be combined with other medications and treatments. Because anti-inflammatory effects and other improvements are occurring within a few days, treating episodic exacerbations, which usually take from one week to several months, patients experience an effective reduction in unpleasant or even unbearable symptoms during the course of treatment with CRF.

Brief description of the drawing

Figure 1 is a perspective view illustrating two Liuiso LEW / N mouse males on susceptible mycobacterium-induced arthritis where 10 paws are in control and 12 paws on Mycobacterium arthritis;

Figure 2 is a graphical illustration of foot padding (one unit corresponding to 0.001 inches) using an arthritic-rat animal model, time-dependent control animals and animals treated with the present invention using two different doses (50 and 100 μ% CRF; injected subcutaneously) , per kilogram of body weight) for which 4 inimas (leg protrusion was measured on the toe, into which mycobacteria were injected (left posterior);

Fig. 3 is similar to Fig. 2, but only illustrates measurements of paws which are opposite to the transverse plane on the side of the paws to which the injections (right posterior) were made;

Figure 4 is a graphical illustration of the time dependence of arthritis signs of the right hind paw from Figure 3;

Fig. 5 is similar to Fig.4, but only a pair of paws (front left and front right) are measured;

Fig. 6, including part A-E, is similar to Fig. 2; only a higher dose of the present invention (100 µg / kg body weight) is administered subcutaneously to the left leg of the mycobacterial affected leg 40 days after the onset of the disease;

Fig. 7, including part A-E, is similar to Fig. 6, only the right leg of the leg opposite to the transverse plane is measured; and

Figure 8 graphically illustrates the dependence of arthritic signs similarly to Figure 4, but the format is similar to Figure 6.

It is possible that the previous reports that CRF is found in the extremities of mammals with inflammatory arthritis and that CRF is a proinflammatory agent are correct because it can be assumed that this peptide may be either pregmatory or anti-inflammatory, depending on the amount of endogenous peptide produced by how the "inflammatory" cell reacts to CRF or when the CRF is formed during the inflammatory response. However, a further description of this work shows that administration of CRF to a patient may be therapeutically effective in stopping episodic exacerbations and chronic inflammatory responses in a dose-dependent manner. Thus, the present invention is directed to the use of CRF for the manufacture of a medicament for the treatment of patients with chronic inflammatory arthritis such as rheumatoid arthritis. The practice of the present invention uses CRF. The term "CRF" includes CRF analogues and CRF-related peptides known in the art. The CRF therapeutic compositions for storage may be in the form of lyophilized adherent particles or aqueous solutions by mixing the CRF with the desired degree of purity with selected physiologically acceptable carriers, additives or stabilizing agents. Acceptable carriers, excipients, or stabilizing agents should be non-toxic to the consumer if the required doses and concentrations are used. Such materials include buffers such as phosphate, citrate and other organic acids; antioxidants, including ascorbic acid; low molecular weight (less than 10 residues) polypeptides; proteins such as serum albumin, gelatin or immunoglobulins. Other components include glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates, including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming contractions such as sodium; and / or nonionic surfactants such as Tween, Pluronics or PEG.

The mode of administration may be any of the known methods, including but not limited to intraarticular, intravenous, intrathecal, subcutaneous, injectable, intranasal, oral or implanted devices. Suitable implants include, for example, gel foam, wax or microparticle-based implants. Preferably, the preparations for topical administration include agents that facilitate transdermal delivery, such as melting agents, or dermal transfer devices, such as well-known targeted delivery systems, and the development of rapidly transmitting dermal transfer technologies.

The therapeutically effective doses of CRF or its analogues for humans according to the present invention are preferably at least about 0.01 µg / kg. For systemic administration (eg, subcutaneously or intravenously), a higher dose is about 0.1 to 50 µg / kg, preferably about 1 to 30 Mg / kg. For topical administration (eg by joining the joints), the preferred dose is about 1 to 100 µg / kg. Administration of CRF doses can be slow, for example by subcutaneous or intradermal injection, or can be injected directly into the painful part of the body.

Therapeutic patients should be supervised by a health care professional, such as a therapist, who can self-inject a therapeutic composition containing CRF, or assign a composition to the patient that he / she may inject (such as pre-filled syringes). Particularly suitable are forms of systemic use, since inflammation occurs in many places. Although the present invention provides the use of CRF, it is contemplated that the use of CRF may be used

As will be illustrated below, it appears to be effective, but the efficacy disappears when treatment is discontinued. CRF and its analogues can be combined with other medicines.

According to the invention, the use of CRF may potentially last up to about three weeks, but side effects (eg Cushing's syndrome) begin to occur after long-term use. Thus, in the practice of the present invention, it is primarily intended to be used for the treatment of acute episodic outbreaks, and CRF can also be started 40 days after episodic exacerbation.

Although peptides are generally water-soluble, if they are typically synthesized, they can be used in the form of pharmaceutically acceptable non-toxic salts such as salts with acids. Examples of salts with acids include hydrochloride, hydrobromide, sulfate, acetate, citrate, benzoate, succinate, maleate, ascorbate, tartrate and the like. In addition to being able to use CRF in any pharmacologically acceptable carrier, it may be incorporated into a liposome, microcapsule, polymer or wax-based and controlled release drug, depending on the mode of administration desired, or may be in the form of tablets, in the form of pills or capsules. CRF or analogues thereof can be used to treat patients with one of the chronic inflammatory arthritis. Chronic inflammatory arthritis is considered to include rheumatoid arthritis, Reiter's syndrome, ankylosing spondylitis, psoriatic arthritis, and inflammatory bowel disease. Treatment with CRF is particularly suitable for episodic exacerbations and provides anti-inflammatory results depending on the dose. Experiments show that regardless of the time of use (e.g., and after about 40 days of episodic exacerbation), using the CRF provided by the invention reduces inflammation and in the presence of a chronic disease; however, when the treatment is stopped, the symptoms of inflammation appear again. Thus, the preparation of the present invention is intended for facilitation rather than full cure. 7

Experimental part

Therapeutic efficacy is demonstrated using an adjuvant arthritis model in which arthritis is injected into the arthritis-bearing Liuiso "LEW / N" male left hind paw with an inoculated Mycobacterium bacterium.

As shown in Fig. 1, about sixteen days after the onset of the disease (i.e., the animals are treated with a saline solution), the paws of the animals are like a shelf 10; however, immediately after the onset of the disease, the paw to which the mycobacteria were injected (and later as described below and the other paws) is the same as the 12 paw, i.e. swelling occurs. Swelling of the foot pad is an ignition signal. It is known that inflammation is accompanied by redness, swelling, heat and pain as a body reaction to injury or other effects. Five days after the onset of the disease, CRF was administered twice daily at doses of 50 µg / kg or 100 µg / kg / kg. Medicinal product no longer authorized twice daily for 20 days. Approximately 16 days after the onset of the disease, the swelling or inflammation of CRF treated animals was relatively small.

Starting from the first day of the drug release (i.e. the fifth day after the injection of dead mycobacteria), the swelling size was determined by a crown, measuring the thickness of the left hind (into which the mycobacteria was applied). This measurement was used to assess the degree of 'primary' or acute inflammation. Later (approximately 10-12 days later), it becomes swollen and becomes inflamed in the right hind paw. This represents a chronic inflammatory response.

Figure 2 shows the left hind paw containing the primary inflammation in rats receiving CRF at 50 / kg / kg twice daily in rats receiving CRF at 100 / kg / kg twice daily; , swelling data (measured for 32 days) .The vertical axis is divided into units where one unit corresponds to 0.001 inches. As can be seen from Fig. 2, the use of CRF according to the present invention provides anti-inflammatory results and is dose-dependent. The three curves shown in Fig. 2 begin 5 days after the mycobacterium injection into the left hind paw. When a dose of 50 / kg / kg CRF is given twice a day, the swelling compared to the control is reduced. A further decrease is observed after injecting 100 µg / kg / kg 8

Fig. 3 is similar to Fig. 2, but only the degree of swelling of the right hind paw, which corresponds to a chronic inflammatory response, is measured. The injection of 50 µg / kg or 100 µg / kg of CRF was started five days after mycobacterial challenge, as in Figure 2, but the right hind paw begins to swell later than the left hind paw illustrated in Figure 2. In addition, the right hind paw is swollen less than the left hind paw. This is self-evident because the right hind paw is not directly affected as the left hind paw. However, as can be seen from around 11 days, the right hind paw begins to ink; however, when the animals received the CRF according to the present invention, the digestion is very low (both at 50 μg / kg and 100 / µg / kg).

Fig. 4 shows the results obtained with animals in the same manner as in Fig. 3, only using an evaluation system in which "0" means that there is no arthritis, and "4" means highly expressed arthritis. Signs of arthritis are assessed by combining factors that include leg paw damage and general symptoms (such as swelling, redness, and form distortion). Fig. 4 shows that both doses of CRF (50 µg / kg and 100 µg / kg) are active in reducing the signs of arthritis, although a dose of 100 µg / kg has a greater effect. Fig. 4 shows the data obtained by measuring the right hind paw.

Figure 5 shows both front paws in the same measurements as in Figure 4. This is also the case in Figure 4 and Figure 5 illustrates more chronic rather than acute situations. However, the onset and other symptoms of the front paws used to obtain Fig. 5 data are further delayed and less pronounced than the right hind paws used to obtain Fig. 4 data. From Fig. 5, it is particularly apparent that the use of CRF according to the present invention is very effective in the treatment of clinical manifestations of chronic inflammatory arthritis.

In Figs. 5 to 5, treatment was started five days after the onset of the disease. Another series of experiments were carried out in the same way as before and the animals are treated according to the method of the present invention for 20 days (100 µg / kg subcutaneously twice daily), only the course of treatment starts 5 days, 10 days, 15 days, 25 days or 40 days after the onset of the disease. The data of these experiments are summarized in Figure 6-8. 9

Fig. 6A-E illustrates measurements of the left hind leg pawl. Part A shows the results of the treatment of control rats and rats that were started 5 days after the onset of the disease (marked with an indelible arrow) and treatment was stopped after 20 days (indicated by a cursor). Thus, at the start of treatment exactly 5 days after the onset of the disease, the thickness of the paw is greatly reduced compared to the control, and the thickness reduction continues. until treatment is stopped; then the swelling starts again and increases until after about 60 days no difference is seen between the treated paw and the control (untreated) paw. Similarly, Part B illustrates data with control and test rats on Day 10 and ended on Day 30, Part C - Treatment started on Day 15 and ended on Day 35, Part D - Treatment started on Day 25 and completed on Day 45 and Part E Treatment started on day 40 and ended on day 60. These data indicate that treatment with CRF significantly decreases paw extinction even in the case of a long-term illness illustrated by Part E. Thus, as shown in FIG. 6, the treatment of the paw to be treated is reduced by the treatment of 40 days after the onset of the disease.

Fig. 7A-E illustrates similar data as described in Fig. 6, but only the swelling of the right hind paw of the animals was measured. This is, as in Figure 3, since the right back of the right hind paw is measured (mycobacterial injection is made into the left hind paw and reflects acute inflammation), the right hind paw reflects the chronic inflammatory responses. These data again show that the use of CRF according to the invention greatly reduces the tendency even in the case of a chronic disease with chronic inflammation.

In experiments similar to the experiments shown in Figures 6 and 7, but in which the degree of arthritis was evaluated by the system of traits described in Figure 4, it is shown that treatment according to the present invention is effective with respect to general features, although perhaps not clearly visible, than by reducing the swelling of the paw. In each of Fig. 8, the start of treatment of the animals in parts A-E is marked with arrows. The end of treatment time is indicated by cursor arrows.

Thus, as shown in Figure 8A, five days after the ingestion of dead mycobacteria in the left hind leg of the animal, CRF treatment according to the present invention was started with the right hind paw and treatment continued for 20 days. In the course of treatment, signs of arthritis decreased as compared to control, but these 10 signs returned to strength after discontinuation of treatment. Thus, it is appropriate to use CRF in therapeutic practice rather than prophylaxis. However, it is important to note (this is illustrated in Figure 8D) that CRF treatment even after 25 days is still effective.

It should be recalled that the foregoing description of the invention, the most suitable embodiments and examples of the present invention are intended to illustrate the invention and do not limit the scope of the invention as defined by the appended claims.

Claims (10)

Use of Corticotropin Releasing Factor (CRF), including its salts, analogues, or analogues thereof, in the manufacture of a medicament for the treatment of patients with one of the chronic inflammatory arthritis. Use according to claim 1, characterized in that the salts of CRF or its analogues are salts of inorganic or organic pharmaceutically acceptable acids. Use according to claim 1 or 2, characterized in that the medicament produced > is for the treatment of rheumatoid arthritis. Use according to any one of claims 1 to 3, characterized in that the manufactured drug is for reducing episodic exacerbations of inflammation. Use according to any one of claims 1 to 4, wherein the medicament is effective when used up to 40 days after the onset of inflammation. Use according to claims 1-5, characterized in that the medicament produced is used for a maximum of three weeks without interruption. Use according to claims 1-6, characterized in that the manufactured drug is administered topically, intraarticularly, intravenously, intrathecally, subcutaneously, intranasally, orally or via implanted devices. Use according to claims 1 to 7, characterized in that the single dose of the manufactured drug is at least 0.01 µg / kg of active ingredient per kilogram of body weight. 9. Use according to claims 1 to 7, characterized in that the single dose of the manufactured drug for local injection is from 1 to 100 / Ag of the active ingredient per kilogram of body weight. 10. A pharmaceutical composition for the treatment of chronic inflammatory arthritis, characterized in that it contains an effective amount of the salts of CRF and / or its salts, analogues or analogues thereof, and a pharmaceutically acceptable carrier.