JP2002515459A - Method for treating asthma and other allergic conditions and use of a composition comprising a TNF-RII (P75) agonist - Google Patents

Abstract

  (57) [Summary] The present invention is directed to methods of treating asthma, allergies, and conditions of Th2-mediated inflammation by administering to a human or animal a product having TNF-RII (p75) agonist properties. Use of a product having TNF-RII (p75) agonist properties for the manufacture of a pharmaceutical composition for the treatment of asthma, allergy and Th2-mediated inflammation symptoms in humans or animals, and lung inflammation Also provided by the invention is a method of screening for a product having TNF-RII (p75) agonist properties, comprising monitoring inhibition of lung inflammation that has occurred in a human or animal having TNF-RII.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001] BACKGROUND Asthma invention is a specific allergen, exercise, complex inflammatory disease of the airways that is characterized by recurrent deterioration due to exposure to cold air or stress (recurrent exacerbations). Characteristics of inflammation associated with asthma symptoms are activated eosinophils, increased sensitivity of the airway to nonspecific stimuli (airway hyperresponsiveness: AHR), edema, the presence of mucus hypersecretion and cough. This inflammation progression is believed to be mediated in part by the generation and activation of Th2-type lymphocytes that secrete various cytokines. In addition, allergic conditions other than asthma may result in similar production and activation of Th2-lymphocytes and secretion of cytokines, such as rhinoconjuctiv
itis), conjunctivitis, rhinitis, dermatitis, gin machine, anaphylaxis, and others.

[0002] A number of therapeutic agents have been used to treat asthma, notably including steroids and β-2 agonists. Despite these treatments, asthma morbidity and mortality remain high and even increasing. There is certainly a need for drugs that can rapidly reverse the inflammatory response seen in asthma, as current treatments are only effective to a small extent. The same is said from other allergic diseases. For more information on potential treatment approaches for asthma, see Bousquet J. et al. Et al.,
(Clinical and Experimental Allergy, 25 (2), 39-42, 1995).

The tumor necrosis factor TNFα plays a key role in the cytokine network with respect to the pathology of many infectious and inflammatory diseases. TNFα was purified, sequenced, and cloned in the mid-1980s. Since that time, several biological properties of this cytokine have been demonstrated.

[0004] Human TNF is synthesized as a pro-protein containing 233 amino acids with a molecular mass of 26 kDa. This protein is a specific metalloprotease (TNFα-converting enzyme TACE
) Yields a 17 kDa monomeric form containing 157 unglycosylated amino acids. Under physiological conditions, TNFα forms non-covalently bound corn-form homotrimers.

[0005] The action of TNF is transmitted through the cross-linking of the membrane-bound receptor molecules TNF-receptor I (TNF-RI, p55) and TNF-receptor II (TNF-RII, p75). The extracellular portions of both TNF receptors can be released, and these soluble receptors retain the ability to bind to TNF. After binding to its membrane-bound receptor, TNF mediates a variety of effects in other organs and tissues.

[0006] Although some cell types produce TNF, the primary source of this cytokine is monocyte /
Macrophages. TNF induces many of the proinflammatory changes in endothelial cells, including cytokine production, expression of adhesion molecules, and procoagulant release. These changes may lead to septic shock.

[0007] The effects of TNFα are known to promote the symptoms of asthma; thus, another group has found that blocking the action of soluble, circulating TNFα provides a means to alleviate these symptoms Make sure you do. This problem has been addressed by another approach in the published patent literature.

For example, WO 97/41895 relates to the treatment of asthma by administering a preparation of a chimeric protein comprising a soluble part of TNFRI (p55) fused to an IgG. This composition is:
Claims to block the action of TNFα and restore inflammation in the lungs. However, prolonged external administration of the soluble portion of the TNFRI receptor can cause changes in the organism's immune response.

[0009] According to Grell et al. (Cell, 83, 793-802, 1995), membrane-bound TNF (mTNF) is superior to soluble TNFα in inducing p75 receptors in a variety of systems, such as thymocyte proliferation. It is known that In particular, the authors provide evidence that mTNF is the first bioactivator of TNF-RII, and that TNF-RII
Controls the local response pattern. The proposed model has mTNF
If it is a physiological precursor of sTNF, it can be an early and very effective stimulus because it can induce independent signaling pathways in each of the TNFRs.

[0010] Yoshida et al. (Clin. Exp. Immunol., 106 (1) 73-78, 1996) reported that the level of soluble TNFRs with specific inhibitory activity on TNFα was increased in patients with bronchial asthma during asthma challenge. High in serum, indicating that it may contribute to controlling TNFα production and the development of allergic inflammation.

The role of each of the p55 and p75 receptors in mediating and modulating the biological effects of TNF has been studied by generating and testing mice genetically deficient for these receptors. (Peschon J. et al., J. Immunol., 160, 943-952,
1998). Heat-inactivated actinomycetes M. Lung inflammation after intranasal instillation of faeni Ags-
The Th1 inflammatory pathway model was tested in mice lacking p55 or p75 or both. Some selective deficiencies in the inflammatory response lack p55 or p75 and p55
Are observed in mice lacking both, and are not observed in mice lacking p75. In the latter model, the activity of p55 is not reduced by the absence of p75,
Disputes a physiological role for p75 as an essential element of p55-mediated signaling. In contrast, exacerbated lung inflammation and dramatically increased serum TNFα levels in mice lacking p75 suggest a dominant role for p75 in suppressing TNF-mediated inflammatory responses. Thus, p55 and p75 TNF receptors appear to represent an equilibrium system for TNF action. Conversely, deficiency in p75 activation or p75 receptor leads to exacerbated lung inflammation. These results suggest a role for endogenous p75 in down-regulating the pulmonary inflammatory response mediated by Th1-type inflammatory pathways.

[0012] DETAILED DESCRIPTION OF THE INVENTION This application, binding of an agonist of TNF-RII (p75) receptors, a Th2- mediated reduction of inflammation, is notable allergic bronchial asthma and other allergic symptoms It is based on the hypothesis that it attacks TNF-RI (p55) -independent signals on cells, which causes a decrease. This hypothesis is based on the findings reported in instances where selective TNF-RII (p75) activation has a protective role in lung inflammation and hyperresponsiveness in a mouse model of allergic bronchial asthma.

Accordingly, a primary object of the present invention is a method of treating allergic bronchial asthma by administering an effective amount of a product having the properties of a TNF-RII (p75) agonist.

It is a further object of the present invention to provide other allergic conditions such as rhinoconjunctivitis, conjunctivitis, rhinitis, dermatitis, gin machine by administering an effective amount of a product having the properties of a TNF-RII (p75) agonist. , Anaphylaxis, and others.

[0015] Yet another object of the present invention is a method of treating Th2-mediated inflammation by administering an effective amount of a product having TNF-RII (p75) agonist properties. The term "product having the properties of a TNF-RII (p75) agonist" means any molecule capable of binding and activating a receptor, for example, a monoclonal antibody, for example by Bigda et al. The described monoclonal antibodies (J. Exp. Med., 180, 445-460, 1994), and in particular epitope A of the p75 receptor
, B and C, as well as the monoclonal antibodies described in WO 94/09137. The term refers to polyclonal antibodies, proteins, peptides, peptoids, nucleic acids, analogous chemicals of peptides and nucleic acids, and any organic molecule having a molecular weight similar to other organic molecules having pharmacological properties, preferably from about 100 to 30,000 daltons. Also encompasses molecules. The term also includes prokaryotic or eukaryotic, viral, or any other TNF-RII (p75) agonist particles.

[0016] Preferably, the TNF-RII (p75) agonist used in the present invention is TNF-RI (p55)
Has essentially no agonist properties. An "effective amount" is sufficient to affect the course and severity of allergic symptoms, bronchial asthma or Th2-mediated inflammation, and to ameliorate the patient's symptoms, leading to amelioration or remission of the disease. (P75) means the amount of agonist. The effective amount will depend on the route of administration and the condition of the patient.

In the present invention, administration of the product having TNF-RII (p75) agonist properties can be enteral, parenteral, or preferably topical, for example in an aerosol formulation. In a first embodiment of the present invention, a product having TNF-RII (p75) agonist properties is used to attack or exacerbate asthma, allergy, or Th2-mediated inflammation.
), The product is administered in an amount sufficient to mitigate the aggressive or exacerbating effects.

In another embodiment of the present invention, a product having TNF-RII (p75) agonist properties is used to prevent or delay the onset or onset of the symptoms prior to the onset or onset of the symptoms. In sufficient quantities, the source asthma, allergy, or Th
Administered to humans or animals with symptoms of 2-mediated inflammation.

[0019] It is a further object of the present invention that it comprises modifying the equilibrium between TNF-RII (p75) and TNF-RI (p55) effectors, asthma, allergy or Th2-mediated inflammatory conditions in humans or animals. How to treat

[0020] Still another object of the present invention is to provide a pharmaceutical composition for treating allergic bronchial asthma, allergic conditions, and conditions of Th2-mediated inflammation, with a pharmaceutically acceptable carrier. Use of products having TNF-RII (p75) agonist properties.

“Pharmaceutically acceptable” is meant to include any carrier, and is a carrier that does not interfere with the effectiveness of the biological activity of the active ingredient and is not toxic to the host to which it is administered. For example, for parenteral administration, a product having TNF-RII (p75) agonist properties may be formulated in a unit dosage form for injection in vehicles such as saline, dextrose solution, serum albumin and Ringer's solution. Good. All modes of parenteral administration are suitable, including intravenous, intramuscular and subcutaneous administration.

In addition to a pharmaceutically acceptable carrier, the compositions of the present invention may also include minor amounts of additives, such as stabilizers, excipients, buffers and preservatives. Combinations of one or more pharmaceutically active products are also possible.

A further object of the present invention relates to the use of a product having TNF-RII (p75) agonist properties to modify the equilibrium between TNF-RII (p75) and TNF-RI (p55) effectors.

Yet another object of the invention is to screen for products having TNF-RII (p75) agonist properties, including monitoring the inhibition of lung inflammation occurring in humans or animals with lung inflammation. About the method. Inhibition of pulmonary inflammation can be measured by measuring airway resistance indirectly or by any other suitable means, such as during cellular broncho-alveolar lavage or in any other suitable body fluid, such as eosinophils. ) And soluble (leukotriene) marker levels.

In addition, measuring the level of cellular (eosinophil) and soluble (leukotriene) markers during broncho-alveolar lavage or in any other suitable body fluid of a human or animal is useful for the human or animal. May help establish an appropriate dosage of a product having TNF-RII (p75) agonist properties.

The present invention will now be illustrated by the following examples, with reference to the drawings described below. DESCRIPTION OF THE FIGURES FIG. 1 shows the lung reactivity of OVA-treated BALB / c mice that received increased doses of MCh through a nebulizer.

FIG. 2 shows that p55TNFR ^{− / −} and WT mice after OVA-sensitization cannot show high lung reactivity. FIG. 3 shows total cell counts and differential cell counts in BAL fluid of p55TNFR ^{− / −} and WT mice after OVA-sensitization.

FIGS. 3A (total cell count in BAL) and 3B (fraction of cell count in BAL) show total cell count and count in BAL fluid in p55TNFR ^{− / −} and WT mice after OVA-sensitization. Shows cell counts.

FIG. 4 shows the high reactivity of p75TNFR ^{− / −} and WT mouse lungs after OVA-sensitization. 5A (total cell counts in BAL fluid) and 5B (fractionated cell counts in BAL) show OV
A- Total and differential cell counts in BAL from p75TNFR ^{− / −} and C57BL / 6 WT mice after sensitization.

Table 1 shows the TNFα − / − and tmTNTN tested for pulmonary hyperreactivity and inflammation.
Shows a summary of data obtained using Fk / l mice. Examples Materials and Methods Animals. Female BALB / c or C57BL / 6 mice (6-8 weeks old) were placed in Centre d'Elevage
Janvier, Le Genest Saint-Isle, purchased from France. p55-deficient mice were obtained from Pf
Obtained from Dr. effer (Munich); p75-deficient mice were purchased from Jackson Laboratory. TNFα ^{− / −} and tm TNFk / l mice were isolated from G. Obtained from Kollias (Athens).
Wild-type mice (C57BL / 6) of the same genetic background, gender and age were used as controls.

Sensitization and intratracheal challenge with ovalbumin. Hessel et al., ｛Hessel. va
n Oosterhout, et al. Sensitization was performed using the method described by 1993 ID: 17｝. 10 mg ovalbumin (OVA; A-5503, Sigma Chemical Co., 0.1 ml NaCl)
Mice were immunized every other day for 14 days by intraperitoneal (ip) injection of (St. Louis, MO) (0.9% mass / volume). Sham-sensitized mice received 0.1 ml of NaCl using the same protocol.

Mice were challenged 40 days after the start of parenteral sensitization with an intranasal infusion of 50 ml of a 0.3 mg / ml solution of OVA. For this, mice were anesthetized with 2% inhaled FORENE ^™ (Isofuran, Abbott, Cham, Switzerland). Control mice were PBS only 50
μl was processed. This procedure was repeated daily for 5 days. Lung hyperresponsiveness was assessed 24 hours after the last challenge, 2 days after bronchoalveolar lavage (BAL) was performed.

Bronchoalveolar lavage and counting of fractionated cells. Three days after the third antigen challenge,
Mice were anesthetized with urethane and the trachea was cannulated. BAL fluid was collected by four repeated washes of 0.4 ml of saline solution injected into the lungs through the trachea. Differential cell counting was performed on cytospin preparations stained with Diff-Quick ^™ (Baxter Diagnostics, Dudingen, Switzerland). 3 per sample
Using two fields, 200 cells were counted per field.

Evaluation of allergen-induced airway hyperresponsiveness. Airway responsiveness was measured by recording respiratory pressure curves by whole body blood flow recordings in response to inhaled MCh (Aldrich-Chemie, Steinhein, Germany) at the concentrations indicated in the figure legend (Buxco (Registered trademark), EMKA Technologies, Paris, France)
. The duration of nebulization was 20 seconds. After each dose, lung hyperresponsiveness returned to baseline. This was typically in the order of 15 minutes. This method allowed the measurement of spontaneous respiration in unrestrained mice. Airway responsiveness is enhanced with arrest
hanced pause (Penh), expressed as a calculated value and correlated with measurements of airway resistance, impedance and intrapleural pressure in the same mouse. Penh = (Te / Tr-1)
X Pef / Pif (Te = exhalation time, Tr = relaxation time, Pef = peak expiratory flow, Pif = peak inspiratory flow) x 0.67 = constant established by the manufacturer. Tr = relaxation time. the time is,
It was taken in terms of the pressure of the box on the change from the maximum to the percentage of the maximum specified by the user. In this study, Tr measurements started at maximum box pressure and ended at 40% {Chvatchko, Kosco-Vilbois et al
. 1996 ID: 331｝. Results: A) Neutralization of TNFα FIG. 1 shows the lung reactivity of OVA-treated BALB / c mice that received increased doses of MCh via nebulizer.

Three groups are shown: 1) Control group: Balb / C without antibody treatment 2) Experimental group receiving 50 mg anti-TNFα antibody 2 hours before each challenge 3) Receiving 50 mg isotope control Experimental group.

High responsiveness was monitored by plethysmography and expressed as maximum Penh value over 15 minutes after Mch exposure. p <0.05. The curves show that the Penh values in the anti-TNFα treated group were significantly increased at doses of 1.5 to 6 × 10 ⁻² M Mch compared to untreated and isotope control treated mice, p <0.05.

B) Analysis of High Reactivity and Pulmonary Inflammation in P55TNFR ^{− / −} and P75TNFR ^{− / −} Mice FIG. 2 shows that either NaCl or OVA-sensitization and p55TNFR ^{− / −} and
Figure 5 shows lung reactivity of WT mice. Penh was monitored in response to 6 × 10 ⁻² M Mch as described in Materials and Methods. The results indicate that p55TNFR ^{− / −} animals do not show high lung reactivity to MCh challenge.

FIG. 3 shows that either NaCl or OVA-sensitization and p55TNFR ^{− / −} and
FIG. 4 shows total cell counts and differential cell counts in BAL fluid of WT mice. In FIG. 3, the symbols are as follows: MAC, macrophage; EOS, eosinophil; LYM, lymphocyte; NEUT, neutrophil. Animals challenged with NaCl have no eosinophil infiltrate in their lungs. Eosinophils were abundant in the lungs of C57BL / 6 WT mice after challenge, but not in the lavage fluid of p55TNFR ^{− / −} animals.

The results suggest that p55TNFR ^{− / −} animals do not respond to MCh challenge because they do not contain eosinophils in their BAL. FIG. 4 shows that either NaCl or OVA-sensitization and p75TNFR ^{− / −} and
Figure 4 shows lung reactivity of C57BL / 6 WT mice. Penh was monitored in response to the indicated dosage of MCh, as described in Materials and Methods.

The results show that p75TNFR ^{− / −} mice showed high lung reactivity in a model of allergic asthma. Figure 5 shows that either NaCl or OVA-sensitization and p75TNFR ^-/- and
Figure 4 shows total and fractional cell counts in BAL from C57BL / 6 WT mice.

[0041] Cells were stained with macrophages (MAC) on cytospin preparations after Diff-Quick staining.
), Eosinophils (EOS), lymphocytes (LYM) and neutrophils (NEUT).
Bars represent the mean ± SD of 5 mice per group.

The results indicate that more eosinophils are present in BAL from p75TNFR ^{− / −} compared to C57BL / 6 WT mice, suggesting more inflammation. p75 TNF-R may have a protective role in lung inflammation.

[0043]

[Table 1]

Conclusion: Overexpression of membrane form of TNFα (and soluble TN) in C57BL / 6 x 129sv
Absence of Fα protects animals against MCh-induced lung hyperresponsiveness and reduces lung inflammation, thus suppressing TNF-mediated lung inflammation in this model of allergic asthma A dominant role in doing

[Brief description of the drawings]

FIG. 1 shows the lung reactivity of OVA-treated BALB / c mice receiving increased doses of MCh via nebulizer.

FIG. 2 shows that lung reactivity cannot be demonstrated in p55TNFR ^{− / −} and WT mice after OVA-sensitization.

FIG. 3 shows total cell counts and differential cell counts in BAL fluid of p55TNFR ^{− / −} and WT mice after OVA-sensitization. FIGS. 3A (total cell count in BAL) and 3B (fraction cell count in BAL) show total and differential cell counts in BAL fluid in p55TNFR ^{− / −} and WT mice after OVA-sensitization. Is shown.

FIG. 4 shows the high reactivity of p75TNFR ^{− / −} and WT mouse lungs after OVA-sensitization.

FIG. 5A (total cell count in BAL fluid) and 5B (fractionated cell count in BAL) show OV
A- Total and differential cell counts in BAL from p75TNFR ^{− / −} and C57BL / 6 WT mice after sensitization.

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Claims (22)

[Claims] 1. A method for treating an allergic condition in a human or animal, comprising administering to the human or animal an effective amount of a product having TNF-RII (p75) agonist properties. 2. A method for treating a condition of Th2-mediated inflammation in a human or animal, comprising administering to the human or animal an effective amount of a product having TNF-RII (p75) agonist properties. 3. Treatment of bronchial asthma or a pathophysiological equivalent of bronchial asthma in a human or animal, comprising administering to the human or animal an effective amount of a product having TNF-RII (p75) agonist properties. Way to do. 4. The product having TNF-RII (p75) agonist properties is TNF-RI (p55
4.) The method according to any one of claims 1 to 3, wherein the method has essentially no agonist properties. 5. The human or animal suffers from asthma, allergy, or Th2-mediated inflammatory attack or exacerbation, and the product is in an amount sufficient to reduce the effects of said attack or exacerbation. 5. The method according to claim 1, which is administered by
The method described in the section. 6. Prior to the onset of the attack or exacerbation of the symptoms in an amount effective to prevent or delay the onset of the attack or exacerbation, the underlying asthma, allergy,
5. The method of any one of claims 1 to 4, wherein the product is administered to a human or animal having a symptom of Th2-mediated inflammation. 7. The method according to claim 1, wherein the product is administered enterally. 8. The method according to claim 1, wherein the product is administered parenterally. 9. The method according to claim 1, wherein the product is administered topically. 10. The method according to claim 1, wherein the product is a monoclonal antibody or a polyclonal antibody. 11. The method according to claim 1, wherein the product is a protein, a peptide, a peptoid, or a nucleic acid. 12. The method according to claim 1, wherein the product is a peptide analog or a nucleic acid analog. 13. The method according to any one of claims 1 to 9, wherein the product is an organic molecule having a molecular weight similar to other organic molecules having drug properties. 14. The method according to claim 1, wherein the product is an organic molecule having a molecular weight of 100 to 30,000 daltons. 15. The method of any one of claims 1 to 9, wherein the product is a live prokaryotic or eukaryotic cell, virus, or any other TNF-RII (p75) agonist particle. 16. For treating a human or animal asthma, allergy or Th2-mediated inflammatory condition comprising modifying the balance between TNF-RII (p75) and TNF-RI (p55) effectors. the method of. 17. Use of a product having both TNF-RII (p75) agonist properties and a pharmaceutically acceptable carrier for the manufacture of a pharmaceutical composition for the treatment of an allergic condition in humans or animals. 18. A pharmaceutical composition for treating a Th2-mediated inflammatory condition in a human or animal comprising administering to the human or animal an effective amount of a product having TNF-RII (p75) agonist properties. Use of a product having both TNF-RII (p75) agonist properties and a pharmaceutically acceptable carrier for the manufacture. 19. TNF-RII (p75) for the manufacture of a pharmaceutical composition for treating bronchial asthma or a pathophysiological equivalent of bronchial asthma in a human or animal
Use of products having agonist properties. 20. A product having TNF-RII (p75) agonist properties is a TNF-RI (p5
5) Use according to any one of claims 17 to 19, which has essentially no agonist properties. 21. Use of a product having TNF-RII (p75) agonist properties to modify the balance between TNF-RII (p75) and TNF-RI (p55) effectors. 22. A method of screening for a product having TNF-RII (p75) agonist properties, comprising monitoring the inhibition of lung inflammation that occurs in humans or animals with lung inflammation.