Method and preparation for the treatment of inflammatory diseases of the respiratory tract in horses by inhalation therapy with Aviptadil.

Abstract

The present invention relates to a method and a preparation for treating inflammatory diseases of the respiratory tract in horses, in particular equine asthma, by inhalational administration of aviptadil or another receptor agonist of the VIP receptor (VPAC1 and VPAC2 receptors). Vasoactive intestinal peptide (VIP) is a neuropeptide with anti-inflammatory and bronchodilatory properties, which, when synthetically produced (INN = aviptadil), can be used specifically to reduce inflammation and relax the respiratory muscles. In a preferred embodiment, the effect of aviptadil is enhanced by the simultaneous or prior administration of N-acetylcysteine ​​(ACC). Aviptadil is suitable for treating horses because equine VIP and human VIP are sequence-identical.

Description

1. Introduction: The procedure and preparation involve administering a pharmaceutical composition containing aviptadil at a concentration of 0.02–2.0 mg / ml in physiological saline solution. Inhalation is performed via a nebulizer system that generates particles with a size of 0.1–10 µm to ensure effective deposition in the lower respiratory tract. Treatment is administered at least once daily for 7 to 100 days, with a dose of 0.5–5.0 µg of aviptadil per kilogram of body weight per day. Another aspect of the invention is the use of aviptadil in combination with acetylcysteine, a mucolytic agent that supports mucus clearance and increases the effectiveness of the therapy. The composition may also contain stabilizers such as albumin, acetylcysteine, or protease inhibitors to ensure the stability of aviptadil. The procedure offers a targeted, effective and well-tolerated treatment option for horses with chronic respiratory diseases and significantly improves the quality of life of affected animals. 2. Technical field The present patent relates to a method and a preparation for the treatment of respiratory diseases in horses, in particular equine asthma (formerly called heaves), by the inhalational administration of aviptadil. 3. Background of the invention Equine asthma (formerly known as RAO - Recurrent Airway Obstruction or heaves / heaves, the milder form as IAD - Inflammatory Airway Disease) manifests in varying degrees of severity. The most important symptoms are: Respiratory symptoms: Cough – initially dry, later productive, often when trotting or feeding hay. Nasal discharge – usually clear to mucousy, rarely purulent (except in cases of secondary infection). Increased respiratory rate – especially after exertion, in severe cases also at rest. Prolonged, labored exhalation – sometimes with a "double breath". Abdominal breathing – increased use of the abdominal muscles during breathing; in the chronic stage, a so-called "steam groove" develops (a depression along the abdominal muscles). Wheezing, whistling, or rattling sounds upon auscultation (bronchospasm, mucus). General symptoms: Reduced performance – rapid fatigue, reluctance to work. Exhaustion in advanced cases. In severe episodes: shortness of breath (nostrils spread wide, tense posture, possibly sweating). Severity level: Mild form (IAD): often only a slight cough, sometimes reduced performance, otherwise unremarkable. Severe form (RAO): marked shortness of breath, chronic symptoms, irreversible remodeling processes possible. Conventional therapies such as the administration of corticosteroids or bronchodilators often alleviate symptoms, but carry risks of side effects and do not address the underlying pathophysiology. In particular, conventional steroid therapy shows little effect in equine asthma characterized largely by neutrophils. Vasoactive intestinal peptide (VIP) is a neuropeptide agent known to possess anti-inflammatory and bronchodilatory properties. It promotes the release of nitric oxide (NO) and modulates the immune response in the airways, thereby reducing mucus production, relaxing airway muscles, and minimizing the inflammatory response. Vasoactive intestinal peptide (VIP) binds to two main receptors, both belonging to the family of G protein-coupled receptors (GPCRs, class B): VPAC1 receptor (Vasoactive Intestinal Polypeptide Receptor 1). VIP has a high affinity for both VIP and PACAP (Pituitary Adenylate Cyclase-Activating Peptide). It is widely expressed, e.g., in the lungs, intestines, thymus, and spleen. Activation leads primarily via Gs protein → adenylate cyclase → cAMP to smooth muscle relaxation, vasodilation, and anti-inflammatory effects. VPAC2 receptor (Vasoactive Intestinal Polypeptide Receptor 2) Also a high affinity for VIP and PACAP. Found primarily in the lungs, pancreas, brain, and immune cells. Via similar signaling pathways (Gs → cAMP), it mediates circadian rhythms (in the suprachiasmatic nucleus), bronchodilatory and immunomodulatory effects. Additionally, VIP can also bind to the PAC1 receptor (PACAP Type I receptor) with lower affinity, which primarily prefers PACAP as a ligand. VIP (Vasoactive Intestinal Peptide) VPAC1, VPAC2 (=equal strength), weaker PAC1 endogenous neuropeptide vasodilation, immunomodulation, intestinal motility PACAP-27 / PACAP-38 VPAC1, VPAC2 (good), PAC1 (strongly preferred) endogenous peptide neuroprotection, circadian rhythm, insulin secretion Ro 25-1553VPAC2-selective synthetic cyclic VIP analogue bronchodilator for asthma and COPD (clinical studies) BAY 55-9837VPAC2-selective synthetic VIP analogue: Promotes insulin secretion, diabetes therapy (clinical studies) Lys15, Arg16-Leu27-VIP(1-7)-GRF (8-27)VPAC-active (non-highly selective) hybrid peptide research, signaling pathway studies Other stabilized VIP analogs VPAC1 / VPAC2 (depending on design) modified peptides Research: anti-inflammatory, pulmonary hypertension, immunotherapy There are various VIP receptor agonists, almost all of which are based on the peptide sequence of the VIP peptide. These were primarily developed with the aim of improving their plasma half-life. However, in clinical trials, these compounds were unable to produce a therapeutic effect in human asthma that was superior to standard glucocorticoid treatment. Therefore, it is particularly surprising that such a pronounced therapeutic effect was observed in equine asthma, far surpassing the effects of both inhaled and intravenously administered glucocorticoids. Since only horses considered to have exhausted all other treatment options were treated, a direct comparison with glucocorticoid therapy could not be made. However, the surprisingly strong effect of inhaled Aviptadil demonstrates that its mechanism of action extends well beyond its bronchodilatory effect and exhibits efficacy that significantly exceeds the half-life of Aviptadil. Steroids versus new therapeutic approaches in equine asthma For decades, the treatment of equine asthma has relied primarily on the administration of glucocorticoids. These medications have a strong anti-inflammatory effect and can significantly improve lung function in the short term. However, their use is associated with considerable disadvantages: Side effects: Systemic treatment with dexamethasone, prednisolone, or triamcinolone increases the risk of serious complications such as laminitis, immunosuppression, increased susceptibility to infections, or metabolic disorders. No causal therapy: Glucocorticoids suppress the inflammatory response but do not eliminate mucus buildup or airway hypersensitivity. Long-term problems: Continuous or repeated administration is hardly practical due to the side effects, so relapses are common once the medication is discontinued. Aviptadil as an innovative therapeutic approach Aviptadil, a synthetic analogue of vasoactive intestinal peptide (VIP), offers new perspectives in this area. VIP occurs physiologically in the airways and exerts several beneficial effects there: bronchodilation through direct relaxation of the smooth muscle of the airways, independent of β-receptors; anti-inflammatory effect through modulation of cytokine release and inhibition of neutrophilia; and improvement of mucociliary clearance, which facilitates the removal of mucus. Unlike glucocorticoids, Aviptadil targets physiological regulatory mechanisms without causing the same systemic side effects. Surprisingly, it was also observed that the positive effect on the health and constitution of the horses continued even after treatment with Aviptadil was completed. Combination with expectorants An additional benefit results from the combination with acetylcysteine ​​(ACC): ACC has a mucolytic effect, cleaves disulfide bridges in bronchial mucus and thereby reduces its viscosity. In combination with Aviptadil, the mucus is liquefied and at the same time the respiratory muscles are relaxed, making effective cleansing (coughing up) of the bronchi possible. While glucocorticoids remain effective in the short term for equine asthma, their side effects and lack of causal action remain a major problem. Aviptadil, especially in combination with ACC, offers a promising therapeutic approach that addresses inflammation, bronchospasm, and mucus problems – and does so with a significantly more favorable side effect profile. Another positive influence of ACC on the formulation is the improved stability of the Aviptadil solution due to the antioxidant effect of ACC. 4. Description of the invention This patent describes a novel method and preparation for the use of synthetically produced VIP (aviptadil) in inhalation therapy for horses to effectively alleviate the symptoms of equine asthma and improve lung function, particularly in cases where conventional steroid therapy is unsuccessful. In a preferred embodiment of the invention, the aviptadil is provided in a buffered solution with acetylcysteine. 4.1. Objective of the invention The invention aims to provide a therapeutic method and preparation with at least one of the following functions: reduction of airway obstruction, inhibition of the inflammatory response in the bronchi, regulation of mucus production, long-term improvement of the quality of life of horses with equine asthma. This is especially true for horses suffering from the severe form of equine asthma, RAO. 4.2. Procedure The procedure involves the inhalational administration of a pharmaceutical composition containing aviptadil (or another VIP receptor agonist) using a suitable nebulization system. Steps of the procedure and preparation: Production of the pharmaceutical composition: The solution contains Aviptadil at a concentration of 0.02 - 2.0 mg / ml, dissolved in physiological saline solution with a pH between 7.0 and 7.4. Optional additives: Stabilizers such as albumin, acetylcysteine ​​or protease inhibitors to ensure stability. Other suitable buffer solutions are known to those skilled in the art and have been discussed in detail in the literature. Application method: The medication is administered using a specialized inhaler for horses, which generates particle sizes of 1-5 µm to ensure optimal deposition in the lower respiratory tract. Inhalation duration: 3–15 minutes per session, at least once daily for a period of 7–100 days. In a preferred embodiment, a course of treatment lasts 30 days with one session per day. The treatment can be repeated several times if necessary. Dosage: The preferred dose is 1.0–4.0 µg of aviptadil per kg of body weight per day, divided into the number of inhalation sessions. The dose can be increased if necessary. Treatment monitoring: Regular monitoring of respiratory rate, mucus production and lung function by veterinarians. Inhalation with the Flexineb® equine inhaler is a simple and effective method for administering medication or saline solution directly into a horse's airways. To ensure optimal use, a few steps should be followed. First, the device is prepared. This includes fully charging the battery and checking the individual parts, such as the nebulizer head, the mask, and the tubing, for cleanliness and functionality. Remove the Aviptadil prescribed by the veterinarian (daily dose 1 × 0.8 mg Aviptadil in 5 ml of 0.9% NaCl, aseptically prepared in an ampoule bottle) from the refrigerator approximately 3 hours before inhalation so that the medication can reach room temperature. Pour the medication into the medication cup, which is then tightly sealed. Before the inhalation begins, the horse should be standing in a quiet place, such as the grooming area or its stall. The mask is carefully placed over the mouth and nostrils and secured so that it fits snugly but allows the horse sufficient freedom of movement and does not pinch. It is important that the horse can breathe freely and is comfortable with the mask on. After switching on the Flexineb®, the nebulizer automatically begins to produce a fine aerosol mist, which the horse inhales through the mask. Depending on the setting, a session lasts approximately three to fifteen minutes. During this time, the horse should stand as still as possible; gentle leading at a walk is also possible. After inhalation, the device is switched off and the mask removed. The medication cup and membrane should be rinsed thoroughly with water immediately to prevent residue buildup. Harsh cleaning agents should be avoided. If necessary, mild dish soap can be used, which should then be rinsed with water. All parts must dry completely before being reassembled. To ensure the device remains functional for a long time, regular maintenance is necessary. It should be rinsed after each use, and disinfected at regular intervals with a suitable disinfectant is also recommended. Since the membrane is a consumable part, it must be replaced when its performance deteriorates or according to the manufacturer's instructions. The mask should also be cleaned regularly with a mild cleaner to prevent bacterial growth. For practical application, it is helpful to slowly accustom the horse to the mask if it has no prior experience with it. 5. Advantages of the invention Targeted effect: Inhalational application ensures that Aviptadil reaches the deep airways directly without causing systemic side effects. Anti-inflammatory: Aviptadil reduces the release of pro-inflammatory cytokines, thereby preventing the worsening of inflammation. Due to its anti-inflammatory effect, treatment over a month or longer leads to a sustained reduction in airway hyperreactivity. Bronchodilation: Aviptadil promotes relaxation of the smooth muscles of the airways, thus improving breathing. Long-term application: Due to its good tolerability, Aviptadil can also be used over longer periods of time. Inhalation with Aviptadil is particularly effective when standard medication fails, especially in horses with RAO. The additional use of acetylcysteine ​​significantly improves the accessibility of Aviptadil to the target tissue (mucolytic effect). 6. Examples and Results Example 1: Summary of Individual Healing Attempts Over a period of nine months, 10 horses with diagnosed equine asthma that had not responded to other treatments were each treated once daily for four weeks with Aviptadil (0.8 mg / 5 ml). The Flexineb® inhaler was used. After 7 days, a significant improvement in respiratory rate and a reduction in coughing were observed. After 30 days, lung function (measured as pO2) had improved by an average of 16.7%. A significant reduction in respiratory hyperreactivity was observed in approximately 70% of the horses (behavior after physical exertion, willingness to perform, ability to regenerate). Example 2: Comparison with corticosteroids Compared to standard treatment with inhaled corticosteroids, horses treated with Aviptadil showed a significantly greater improvement in symptoms, but without the side effects such as muscle wasting or immunosuppression. In one horse with corticosteroid resistance, Aviptadil resulted in a marked improvement in symptoms (lung function, e.g., pO2). After treatment with dexamethasone (6 months), this horse still had a respiratory rate of 27 breaths / minute. After administration of Aviptadil (0.8 mg / 5 ml per day for 4 weeks), a rapid improvement to 11 breaths / minute occurred, and the pO2 increased from 69% to 84% within one month, even though the steroids had been discontinued! Example 3: Preparation for a powder inhaler Composition (per dose): Aviptadil: 20 mg Acetylcysteine ​​200 mg Lactose monohydrate: 200 mg Microcrystalline cellulose: 80 mg Magnesium stearate: 4 mg The following example describes a formulation for an inhaled medication containing aviptadil. This formulation allows for targeted delivery of the active ingredient to the airways to achieve maximum respiratory effect. Aviptadil, the main ingredient of the medication, is used in a 5 mg dose. It is a powder with high purity and potency. The formulation also contains excipients to improve the stability and handling of the medication. Lactose monohydrate is used as a carrier to ensure even distribution of the active ingredient. Microcrystalline cellulose serves as a bulking agent to stabilize the formulation. Magnesium stearate is used as a lubricant to ensure smooth processing of the medication during manufacturing. The medication is produced in powder form for inhalation. A special inhalation device is used for the horse to ensure the correct dose of the powder is administered. It should be noted that the dose is higher compared to liquid inhalation. The dosage and frequency of inhalation are determined according to the severity of the illness and the individual needs of the horse. It is recommended that a veterinarian or a specialist in inhalation medications determine the exact dosage and application of the medication. The described formulation offers a convenient and effective way to administer Aviptadil for the treatment of equine asthma via inhalation. Targeted delivery to the airways allows for optimal efficacy in slowing disease progression and alleviating patient symptoms. Example 4: Preparation without acetylcysteine ​​for a nebulizer, e.g. Flexineb® 0.9% (w / v) NaCl 0.02% (w / v) AviptadilAqua ad iniectabilia: ad 50 ml / batch Example 5: Preparation with acetylcysteine ​​for a nebulizer, e.g. Flexineb® 0.9% (w / v) NaCl0.02% (w / v) Aviptadil10% w / v Acetylcysteine ​​(ACC)pH 7-8, adjusted with 10% NaOHAqua ad iniectabilia: ad 50 ml / batch Example 6: Case study of horse 1 without ACC Horse 1 was treated once daily for four weeks with Aviptadil (0.8 mg / 5 ml, see Example 4). The Flexineb® inhaler was used. This horse was diagnosed with a severe form of equine asthma (RAO). The following values ​​were recorded at the beginning and end of the treatment (see Fig. 1): Oxygen saturation: pO2 - improvement from 60% to 81%. Respiratory rate: breaths / minute - reduction from 20 to 14.5 breaths per minute. Motivation: 0-10 (AU) - very significant improvement from 4 to 8.2. Motivation: 1 = no desire to exercise / 10 = very high desire to exercise. Example 7: Case study of horse 2 with Aviptadil plus ACC Horse 2 had a severe mucus buildup, so treatment with standard inhalation medications was not very promising. Therefore, treatment with Aviptadil (0.8 mg / 5 ml with 10% ACC, see Example 5) was administered once daily for four weeks. The Flexineb® inhaler was used. The following values ​​were recorded at the beginning and end of the treatment (see Fig. 2): Oxygen saturation: pO2 – improvement from 66% to 88%. Respiratory rate: breaths / minute – reduction from 24 to 11.2 breaths per minute. Performance level: 0-10 (AU) – extremely significant improvement from 2 to 10. After the treatment ended, the horse's health continued to improve. Even six months after discontinuing the medication, no relapse to the initial condition was observed. Surprisingly, no rebound effect, as seen with steroids, was observed. Example 8: Use of Ro 25-1553 for the treatment of horses 0.9% (w / v) NaCl0.02% (w / v) Ro 25-1553Aqua ad iniectabilia: ad 50 ml / batch Horse 3 was treated once daily for four weeks with Ro 25-1553 (0.8 mg / 5 ml). The Flexineb® inhaler was used. The effect was comparable to that of Aviptadil from Example 6. Example 9: Combination of Aviptadil and a glucocorticoid For inhalation with Aviptadil according to Example 6, a horse that had previously responded only moderately to fluticasone propionate alone and subsequently to Aviptadil alone (both via inhalation) was treated with additional inhalation of fluticasone propionate: 2,000 µg of fluticasone propionate was administered once daily for 10-15 minutes via the Flexineb® inhaler. The combined administration produced an additive effect, resulting in a significant improvement in the horse's health over the 30-day treatment period. Example 10: The stability of Aviptadil is improved by combining it with ACC: The addition of 1% - 10% ACC, buffered to pH 7.2, improves the stability of 200 µg / ml Aviptadil for inhalation in physiological saline solution, thus extending the shelf life from 6 months at +4 °C to 24 months. The addition of ACC to Aviptadil leads to increased solubility and bioavailability. Furthermore, the addition of ACC improved the homogeneity of the solution due to its mucolytic and interaction-reducing properties. This ensured that Aviptadil remained evenly distributed in the solution, thus maintaining its efficacy over extended storage periods. Aviptadil 0.9%NaCl100 µg / ml93.90%86.50%73.20% Aviptadil + 5%ACC100 µg / ml98.70%97.50%95.10% Stability testing of compositions containing aviptadil (control) and aviptadil + ACC: Stability of the aviptadil solution with and without ACC when stored at 2 to 8 °C. The determination was carried out by HPLC. Both solutions were adjusted to physiological pH. Conclusion: The addition of ACC to an Aviptadil-containing solution significantly contributes to extending the stability of Aviptadil to up to 24 months at the tested temperature of +4 °C. Another function of ACC in the Aviptadil solution is to reduce the binding of Aviptadil to plastic or glass surfaces. This is particularly important when using small volumes. 7. Scope of protection The patent claims: 1. A method for treating equine asthma in horses by inhalational administration of aviptadil – a synthetically produced vasoactive intestinal peptide (VIP) – in a pharmaceutical composition. 2. The specific use of aviptadil for reducing inflammation and bronchial obstruction in equine asthma, particularly in RAO (Recurrent Airway Obstruction). 3. A pharmaceutical preparation containing aviptadil, suitable for treating equine asthma in horses. 4. A pharmaceutical preparation containing, in addition to aviptadil, a mucolytic agent. Preferably a reducing mucolytic agent selected from the thiol group, e.g., acetylcysteine, mesna, carbocysteine. 5. An inhalation therapy system that effectively delivers aviptadil in a particle size of 1–5 µm, particularly to the lower respiratory tract, wherein the nebulization system is a special equine inhaler. One example of such an inhaler is the Flexineb® inhaler. 6.Aviptadil as defined in the invention comprises the synthetic VIP as well as chemical analogues and derivatives, which were developed in particular to increase stability against enzymatic degradation. 8. Conclusion The present invention represents an innovative and effective therapy option for the treatment of equine asthma in horses and opens up new possibilities for the long-term care of horses with chronic respiratory diseases. 9. Description of the illustrations Fig. 1: 1a) Measurement of blood oxygen saturation at the start of treatment with Aviptadil and after 30 days of treatment. 1b) Measurement of respiratory rate at the start of treatment with Aviptadil and after 30 days of treatment. 1c) Measurement of the horse's willingness to perform at the start of treatment with Aviptadil and after 30 days of treatment. 1 = no willingness to perform, 10 = extremely high willingness to perform. Fig. 2: 2a) Measurement of blood oxygen saturation at the start of treatment with Aviptadil and ACC and after 30 days of treatment. 2b) Measurement of respiratory rate at the start of treatment with Aviptadil and ACC after 30 days of treatment. 2c) Measurement of the horse's willingness to perform at the start of treatment with Aviptadil and ACC and after 30 days of treatment. 1 = no willingness to perform, 10 = extremely high willingness to perform.

Claims

Preparation for inhalation containing a vasoactive intestinal peptide (VIP) receptor agonist, characterized in that it is intended for the treatment or prevention of equine asthma. Preparation comprising a vasoactive intestinal peptide (VIP) receptor agonist and a mucolytic agent, characterized in that it is intended for the treatment or prevention of equine asthma. Preparation according to claim 1 or 2, characterized in that the VIP receptor agonist is a natural VIP, a VIP derivative or a synthetic analogue of VIP. Preparation according to claim 3, characterized in that the VIP receptor agonist is aviptadil or a functional derivative or precursor thereof. Method for treating equine asthma, comprising the inhalational administration of a pharmaceutical composition containing aviptadil in an effective amount, wherein the administration is carried out by means of a nebulization system that produces particles with a size of 0.1 to 10 µm, preferably 1 to 5 µm, to ensure deposition in the lower respiratory tract. Method according to claim 5, wherein Aviptadil is administered in combination with a mucolytic agent, in particular acetylcysteine, to promote mucus clearance. Method according to claim 5 or 6, wherein Aviptadil is administered in combination with a glucocorticoid.

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