Use of epidermal growth factor depletion agents in the treatment of chronic obstructive pulmonary disease

Abstract

The present invention relates to the fields of biotechnology and medicine. Specifically, it describes the use of epidermal growth factor (EGF) depleting agents that contribute to the reduction and / or depletion of serum epidermal growth factor levels, which has an impact on the treatment of chronic obstructive pulmonary disease. These agents can be vaccine compositions containing a conjugate between recombinant human EGF and a carrier protein as an active ingredient.

Description

Technical Field

[0001] The present invention relates to the fields of biotechnology and medicine. More particularly, the present invention relates to the use of epidermal growth factor (EGF) depletion agents, specifically, a vaccine composition comprising epidermal growth factor (EGF) and a carrier protein as an active ingredient in the treatment of chronic obstructive pulmonary disease (COPD).

Background Art

[0002] COPD is characterized by the presence of chronic and irreversible airflow obstruction associated with abnormal and chronic inflammatory responses that lead to remodeling of the airways, parenchyma, and pulmonary arteries, and also acts systemically (Fletcher C. et al., (1977) Br Med J. 1(6077):1645-1648). The prevalence of COPD and the mortality due to COPD have increased significantly worldwide, and this disease is currently the fourth leading cause of death in the world (NIH Publication No. 2701A, March 2001, available at www.goldcopd.com). Continuous exposure to tobacco smoke and biomass combustion products in closed environments is (Perez-Padilla R. et al. (1996) Am J Respir Crit Care Med. 154:701-706; Orozco-Levi M. et al. Eur Respir J. (2006); 27(3):542-546). Homozygous alpha1-antitrypsin deficiency is associated with early emphysema in smokers and leads to the development of COPD (Alpha-1-Antitrypsin Deficiency Registry Study Group. (1998) Am J Respir Crit Care Med. 158(1):49-59).

[0003] COPD is the greatest risk factor for lung cancer in smokers, and is a population with a 4.5-fold higher risk of developing this type of cancer. Also, this disease is found in 50-90% of lung cancer patients. Lung cancer is also a major cause of the pathological condition and death of COPD patients (Anthonisen N.R. et al., (2005) Ann Intern Med.; 142:233-239).

[0004] The involvement of EGF and its receptor (EGFR) in the pathogenesis of COPD has been suggested. Positive regulation of EGFR achieved by the addition of EGF and TGFα ligands to airway epithelial cells is known to induce the expression of MUC5AC and MUC2 genes, which are responsible for mucus production in the airways. Expression of most receptors and ligands of the EGF / EGFR system is higher in damaged epithelium than in intact epithelium. Overexpression of EGF / EGFR and / or other ligands of the family can lead to changes in normal airway epithelium, such as metaplasia or epithelial hyperplasia (Willem I. et al. Am J Clin Pathol. (2006) 125:184-192). Human eosinophils induce mucin production by airway epithelial cells via EGFR activation (Burgel PR et al. (2001) J Immunol. 167:5948-5954).

[0005] Currently, treatment for COPD is primarily palliative, meaning it mainly focuses on controlling the clinical symptoms of the disease. Clinical investigations are underway regarding the use of drugs aimed at addressing the etiology of this disease.

[0006] The use of a tyrosine kinase inhibitor called BIBW2948 has been studied in COPD, but the treatment regimens used not only failed to reduce mucus production in patients but also resulted in serious adverse effects such as increased transaminase enzymes. Higher in vivo doses showed an effect on reducing mucus production, but the adverse effects reported in clinical trials allowed for dose escalation (Woodruff W. et al. (2010) Am J Respir Crit Care Med 181:438-445).

[0007] U.S. Patent Application Publication 2011 / 0192396 describes the use of a pharmaceutical composition containing EGF and a buffer or saline solution as a pharmaceutical vehicle suitable for the treatment of COPD. The effects of EGF on mucus hypersecretion, alveolar cell protection, and improvement of pathological tissue symptoms in emphysema models have been demonstrated in in vitro and in vivo preclinical studies.

[0008] It is appropriate to emphasize that no prior technical solutions or scientific publications describe the use of immunotherapeutic agents that cause active immune depletion of serum EGF. The novelty of the present invention lies in providing a treatment for COPD with a less toxic EGF-blocking therapy alternative, consisting of the use of a vaccine composition that induces the production of specific antibodies against EGF. The safety demonstrated by these vaccine compositions in cancer treatment allows for their use in long-term treatment and leads to increased efficacy of these therapies. In the present invention, the inventors achieved improvement in the clinical symptoms of COPD by administering the above-described vaccine composition to patients suffering from this disease. [Overview of the project] [Means for solving the problem]

[0009] In one embodiment, the object of the present invention is the use of an EGF-depleting immunotherapy agent for treating chronic obstructive pulmonary disease. Specifically, the immunotherapy agent refers to a vaccine composition that induces the production of specific antibodies against EGF. These vaccine compositions include, as an active ingredient, a conjugate between recombinant human EGF and a carrier protein which may be cholera toxin B subunit, tetanus toxoid, KLH, and P64k derived from Neisseria meningitidis. Furthermore, the vaccine composition has an adjuvant selected from the group consisting of incomplete Freund's adjuvant, squalene adjuvant, synthetic adjuvant, mineral adjuvant, plant adjuvant, animal adjuvant, particulate protein adjuvant, and liposome.

[0010] In another embodiment, the present invention provides a method for treating a subject requiring such treatment, comprising administering the aforementioned vaccine composition via an intramuscular route, preferably in a dose range of 20 to 70 μL / kg, wherein the first four doses are administered every 14 days, and the remaining doses are administered every 28 days for a minimum period of 6 months. [Modes for carrying out the invention]

[0011] Vaccine composition All vaccine compositions provided in the present invention, used as EGF depletion agents in the treatment of COPD, induce the production of antibodies against EGF that contribute to a decrease and / or depletion of serum EGF levels.

[0012] Examples of the vaccine compositions described above include all those containing a conjugate between recombinant human EGF (rhEGF) and a carrier protein as the active ingredient. The carrier protein is selected from the group consisting of, but not limited to, cholera toxin B subunit, tetanus toxoid, KLH, and P64k derived from Neisseria meningitidis. Furthermore, the vaccine compositions described above include adjuvants selected from the group consisting of incomplete Freund's adjuvants, squalene adjuvants, synthetic adjuvants, mineral adjuvants, plant adjuvants, animal adjuvants, particulate protein adjuvants, and liposomes.

[0013] Treatment method Another embodiment of the present invention provides a method of treatment with the vaccine composition described above. Patients diagnosed with COPD according to the GOLD criteria (Global Initiative Guidelines for Chronic Obstructive Pulmonary Disease) (NHLBI / WHO Workshop Report, NIH Publication No. 2701A, March 2001, available at www.goldcopd.com) are eligible for this treatment.

[0014] According to these criteria, COPD is characterized by the presence of chronic, less reversible airflow obstruction associated with an abnormal inflammatory response. Its main clinical symptoms are cough, sputum production, and shortness of breath or a feeling of suffocation during walking or increased activity levels, which gradually worsen over many years. Airflow obstruction is diagnosed by vital capacity measurement when the ratio of forced expiratory volume (FEV1) to forced vital capacity (FVC) in the first second after a bronchodilator test is less than 0.7 (or below the lower limit of normal for subjects over 60 years of age).

[0015] In this invention, COPD patients immunized with the vaccine composition described herein exhibit clinical benefit. In the context of this invention, the term clinical benefit refers to the fact that treatment with such a drug improves and / or stabilizes at least one of the respiratory clinical symptoms presented by the patient.

[0016] In the context of the present invention, the term therapeutic effect generally refers to a beneficial or desirable effect of treatment, such as improvement or remission of the symptoms of a disease.

[0017] The vaccine composition containing EGF as the active ingredient is preferably administered via the intramuscular route, with the first four doses administered every 14 days, and the remaining doses every 28 days, with an acceptable delay time range of 3 days. The dosage range for administering the above composition is between 20 and 70 μL / kg per kilogram of body weight, or 20 to 70 μg of total protein per kilogram of body weight, or a maximum of 5 mg, more preferably 30 to 60 μg / kg of total protein. The minimum duration of treatment is 6 months. This treatment schedule can be modified in frequency and dosage according to the results obtained and can be optimized considering the antibody titer produced and the improvement and / or stabilization of clinical symptoms.

[0018] The present invention will be further described with reference to the following examples and drawings. However, these examples should not be construed as limiting the scope of the present invention. [Brief explanation of the drawing]

[0019] [Figure 1] Baseline serum EGF levels in patients with grade III / IV COPD (severe and very severe stages according to the GOLD criteria) compared to those treated in patients with untreated and chemotherapy stage III / IV non-small cell lung cancer (NSCLC). [Figure 2] Immunosuppression of serum EGF levels and anti-EGF antibody titers achieved over time in grade III COPD patients treated with CIMAvax-EGF. [Figure 3] Inhibition rate of phosphorylation by anti-EGF antibodies.

[0020] Example Example 1. Both COPD patients and NSCLC patients have high baseline serum EGF levels. Baseline serum EGF levels of samples from groups of patients with grade III / IV COPD and NSCLC patients were determined by ELISA (Gonzalez, E Cet al., J Immunoassay Immunochem. (2016) 16:1-12). Similar levels of serum EGF were found in COPD patients and NSCLC patients (Figure 1).

[0021] Example 2. Treatment with CIMAvax-EGF induces anti-EGF antibodies while inducing immunosuppressive serum EGF and improving the clinical symptoms of COPD. Patients with severe COPD (grade III according to the GOLD criteria) were immunized with 2.4 mg of the active ingredient of the therapeutic vaccine CIMAvax-EGF and a total of 5 administrations were given intramuscularly. Anti-EGF antibody titers were quantified by ELISA (Gonzalez G.A. et al Ann Oncol (1998); 9:431-435). Serum EGF levels during treatment were examined by ELISA (Gonzalez, EC et al., J Immunoassay Immunochem. (2016) 16:1-12).

[0022] Figure 2 shows that immunosuppression of serum EGF levels associated with an increase in anti-EGF antibody titers in patients was achieved during treatment.

[0023] Furthermore, spirometry was performed before starting treatment with the vaccine and after treatment was completed. Table 1 shows the results of FEV1, which is the best indicator of the severity of airflow obstruction. [Table 1]

[0024] As can be seen from the results shown in Table 1, treatment with CIMAvax-EGF improved the clinical respiratory symptoms of the treated patients and reduced the severity according to the GOLD criteria.

[0025] Example 3. Treatment with CIMAvax-EGF induces anti-EGF antibodies that have the ability to inhibit EGF receptor phosphorylation. Patients with severe COPD (grade III according to GOLD classification) were immunized with 2.4 mg of the active ingredient of the therapeutic vaccine CIMAvax-EGF, and a total of 8 administrations were given intramuscularly. The inhibitory capacity of the sera obtained before the first immunization and the fifth immunization was evaluated by Western blotting and incubated with tumor cells expressing the EGF receptor for 1 hour. The AG1478 antibody was used as an inhibition control. Figure 3 shows that the antibodies generated by vaccination inhibit EGF receptor phosphorylation in the presence of EGF (36% inhibition).

Claims

1. A vaccine composition for use in the treatment of chronic obstructive pulmonary disease, A vaccine composition comprising a conjugate between recombinant human EGF and a carrier protein as an active ingredient.

2. The aforementioned carrier protein, - Cholera toxin B subunit - Tetanus toxoid, -KLH and - P64k derived from Neisseria meningitidis A vaccine composition according to claim 1, selected from the group consisting of the following.

3. The vaccine composition is - Incomplete Freund adjuvant, - Squalene-based adjuvants, - Synthetic adjuvants, - Mineral-derived adjuvants, - Plant-derived adjuvants, - Animal-derived adjuvants, - Particle protein adjuvants, and - Liposome The vaccine composition according to claim 1 or 2, further comprising an adjuvant selected from the group consisting of the following.

4. The vaccine composition is - Incomplete Freund adjuvant, - Mineral-derived adjuvants, and - Liposome The vaccine composition according to claim 1 or 2, further comprising an adjuvant selected from the group consisting of the following.

5. The vaccine composition according to any one of claims 1 to 4, wherein the vaccine composition is administered by intramuscular route in a dose of 2.4 mg, with the first four doses administered every 14 days and the remaining doses administered every 28 days for a minimum period of 6 months.

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