RESUMEN
The expression of Bruton tyrosine kinase (BTK) in B cells and innate immune cells provides essential downstream signaling for BCR, Fc receptors, and other innate immune cell pathways. The topical covalent BTK inhibitor PRN473 has shown durable, reversible BTK occupancy with rapid on-rate and slow off-rate binding kinetics and long residence time, resulting in prolonged, localized efficacy with low systemic exposure in vivo. Mechanisms of PRN473 include inhibition of IgE (FcεR)-mediated activation of mast cells and basophils, IgG (FcγR)-mediated activation of monocytes, and neutrophil migration. In vivo, oral PRN473 was efficacious and well tolerated in the treatment of canine pemphigus foliaceus. In this study, we evaluated in vitro selectivity and functionality, in vivo skin Ab inflammatory responses, and systemic pharmacology with topically administered PRN473. Significant dose-dependent inhibition of IgG-mediated passive Arthus reaction in rats was observed with topical PRN473 and was maintained when given 16 h prior to challenge, reinforcing extended activity with once-daily administration. Similarly, topical PRN473 resulted in significant dose-dependent inhibition of the mouse passive cutaneous anaphylaxis IgE-mediated reaction. Multiday treatment with topical PRN473 in rodents resulted in low-to-no systemic accumulation, suggesting that efficacy was mainly due to localized exposure. Reduced skin Ab inflammatory activity was also confirmed with oral PRN473. These preclinical studies provide a strong biologic basis for targeting innate immune cell responses locally in the skin, with rapid onset of action following once-daily topical PRN473 administration and minimal systemic exposure. Dose-dependent inhibition in these preclinical models of immune-mediated skin diseases support future clinical studies.
Asunto(s)
Agammaglobulinemia Tirosina Quinasa , Reacción de Arthus , Anafilaxis Cutánea Pasiva , Inhibidores de Proteínas Quinasas , Enfermedades de la Piel , Animales , Femenino , Humanos , Ratones , Ratas , Administración Cutánea , Administración Oral , Agammaglobulinemia Tirosina Quinasa/antagonistas & inhibidores , Reacción de Arthus/tratamiento farmacológico , Reacción de Arthus/inmunología , Reacción de Arthus/patología , Modelos Animales de Enfermedad , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Anafilaxis Cutánea Pasiva/efectos de los fármacos , Inhibidores de Proteínas Quinasas/administración & dosificación , Piel/efectos de los fármacos , Piel/inmunología , Piel/patología , Enfermedades de la Piel/tratamiento farmacológico , Enfermedades de la Piel/inmunología , Enfermedades de la Piel/patologíaRESUMEN
An allergy may sometimes be very dangerous and one of the main factors responsible for allergy is the complement system which can lead to a life-threatening reaction called anaphylaxis. Cycloxygenase-1 (COX-1), Cycloxygenase-2 (COX-2) and 5-Lipoxygenase (5-LOX) trigger allergic and inflammatory reactions. A number of anti-allergic synthetic drugs are available but are costly and show many side effects. Hence, the ancient traditional system of medication mentioned in Ayurveda finds an edge over various synthetic drugs. Zizyphus mauritiana is referred to as the store house of phytochemicals in Ayurveda. The stem and root barks of Zizyphus mauritiana were dried and powdered under controlled conditions. Extractions of the dried powders were performed separately in different solvents in increasing order of their polarity and were tested for their ability to inhibit the complement system. The aqueous extract of the root bark was found to be more effective in inhibiting the complement system. Fractionation of the aqueous extract resulted in the isolation of the Most Active Fraction (MAF) which inhibited the complement system, COX-1, COX-2, and 5-LOX with IC50 values of 0.006 µg ml(-1), 0.065 µg ml(-1), 0.008 µg ml(-1), and 0.083 µg ml(-1), respectively. The MAF was proven to be successful in down-regulating pro-inflammatory mediators like TNF-α, COX-2, and iNOS when tested on a RAW 264.7 cell line. In vivo, the MAF was found to be preventive against anaphylactic shock and the Arthus reaction, when orally administered daily to Wistar rats. Phytochemical analysis of the MAF has indicated that it is rich in tannins. Results indicate that the MAF, a fraction isolated from the aqueous extract of the root bark of Zizyphus mauritiana, has potent anti-allergic and anti-inflammatory properties.
Asunto(s)
Anafilaxia/tratamiento farmacológico , Antialérgicos/administración & dosificación , Antiinflamatorios/administración & dosificación , Reacción de Arthus/tratamiento farmacológico , Extractos Vegetales/administración & dosificación , Raíces de Plantas/química , Ziziphus/química , Anafilaxia/genética , Anafilaxia/inmunología , Animales , Antialérgicos/química , Antiinflamatorios/química , Reacción de Arthus/genética , Reacción de Arthus/inmunología , Ciclooxigenasa 1/genética , Ciclooxigenasa 1/inmunología , Ciclooxigenasa 2/genética , Ciclooxigenasa 2/inmunología , Femenino , Humanos , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Masculino , Proteínas de la Membrana/genética , Proteínas de la Membrana/inmunología , Ratones , Extractos Vegetales/química , Células RAW 264.7 , Ratas , Ratas Wistar , Factor de Necrosis Tumoral alfa/genética , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Lectins bind to surface receptors on target cells, and activate a cascade of events, eventually leading to altered immune status of host. The immunomodulatory potential of purified lectin from Aspergillus nidulans was evaluated in Swiss albino mice treated intraperitoneally with seven different doses of purified lectin. Lectin prevented BSA-induced Arthus reaction and systemic anaphylaxis. The enhanced functional ability of macrophages was evident from respiratory burst activity and nitric oxide production in splenocyte cultures. Interferon-gamma and interleukin-6 levels were significantly up-regulated in treated groups. Maximum stimulatory effect was observed at the dose of 1.5 mg/kg body weight. Therapeutic potential of A. nidulans lectin was assessed against trinitrobenzene sulfonic acid-induced ulcerative colitis in male Wistar rats. Rats pre-treated with 80 mg/kg body weight of purified lectin intraperitoneally prior to colitis induction showed lesser disease severity and recovery within 7 days, while rats post-treated with the same dose showed recovery in 11 days. The results demonstrate immunomodulatory effects of A. nidulans lectin in Swiss albino mice, resulting in improved immune status of the animals and unfold its curative effect against ulcerative colitis in rat model. This is the first report on immunomodulatory and therapeutic potential of a lectin from microfungi.
Asunto(s)
Anafilaxia/prevención & control , Reacción de Arthus/prevención & control , Aspergillus nidulans/química , Colitis Ulcerosa/tratamiento farmacológico , Proteínas Fúngicas , Factores Inmunológicos , Lectinas , Anafilaxia/inducido químicamente , Anafilaxia/tratamiento farmacológico , Anafilaxia/inmunología , Animales , Reacción de Arthus/inducido químicamente , Reacción de Arthus/tratamiento farmacológico , Reacción de Arthus/inmunología , Bovinos , Colitis Ulcerosa/inducido químicamente , Colitis Ulcerosa/inmunología , Colitis Ulcerosa/prevención & control , Modelos Animales de Enfermedad , Relación Dosis-Respuesta Inmunológica , Proteínas Fúngicas/farmacología , Proteínas Fúngicas/uso terapéutico , Factores Inmunológicos/farmacología , Factores Inmunológicos/uso terapéutico , Interferón gamma/biosíntesis , Interleucina-6/biosíntesis , Lectinas/farmacología , Lectinas/uso terapéutico , Macrófagos/citología , Macrófagos/efectos de los fármacos , Macrófagos/metabolismo , Masculino , Ratones , Micelio/química , Óxido Nítrico/biosíntesis , Ratas , Ratas Wistar , Albúmina Sérica/administración & dosificación , Albúmina Sérica/efectos adversos , Albúmina Sérica/antagonistas & inhibidores , Ácido Trinitrobencenosulfónico/administración & dosificación , Ácido Trinitrobencenosulfónico/efectos adversos , Ácido Trinitrobencenosulfónico/antagonistas & inhibidoresRESUMEN
An alkaloidal component, dehydrocorydaline (DHC) isolated from Corydalis Tuber (tuber of Corydalis turtschaninovii forma yanhusuo), has been screened for activity against types I-IV allergic reactions. In a type I allergic models, DHC at a dose of 0.5 mmol/kg, p.o. inhibited 48 h homologous passive cutaneous anaphylaxis (PCA) in rats, which is related to IgE. DHC also exhibited an inhibitory effect on antigen-induced histamine release from peritoneal mast cells. In a type II allergic model, DHC did not inhibit reversed cutaneous anaphylaxis (RCA) in rats. In a type III allergic model, DHC showed weak inhibition on direct passive arthus reaction (DPAR) in rats. Furthermore, in a type IV allergic model, DHC had inhibitory effects on the induction phase and effector phase in picryl chloride-induced contact dermatitis (PC-CD). These results indicated that DHC not only inhibits antibody-mediated allergic reactions but also influences cell-mediated allergia reactions, and the inhibitory effect of Corydalis Tuber on allergic reactions may be partially attributed to DHC.
Asunto(s)
Alcaloides/farmacología , Antialérgicos/farmacología , Medicamentos Herbarios Chinos/farmacología , Hipersensibilidad/prevención & control , Animales , Reacción de Arthus/inmunología , Reacción de Arthus/prevención & control , Dermatitis Alérgica por Contacto/inmunología , Dermatitis Alérgica por Contacto/prevención & control , Liberación de Histamina/efectos de los fármacos , Hipersensibilidad/inmunología , Inmunoglobulina E/biosíntesis , Masculino , Mastocitos/metabolismo , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos ICR , Anafilaxis Cutánea Pasiva/efectos de los fármacos , Ratas , Ratas WistarRESUMEN
A 50% ethanolic extract of Picrorhiza kurroa Royle ex Benth. (Scrophulariaceae) leaves (PKLE) was found to stimulate the cell-mediated and humoral components of the immune system as well as phagocytosis in experimental animals. PKLE elicited a dose-related increase in SRBC, induced 4 h (early) and 24 h (delayed) hypersensitivity reactions in mice and rats, and horse serum induced Arthus reaction in guinea pigs. It also enhanced the humoral immune responses in mice and rats and phagocytic function of the cells of the reticuloendothelial system in mice. PKLE exhibited no mitogenic activity but augmented the responsiveness of murine splenocytes to T cell mitogens phytohaemagglutinin (PHA) and concanavalin A (Con A) and B cell mitogen lipopolysaccharide (LPS E. coli).
Asunto(s)
Extractos Vegetales/inmunología , Animales , Formación de Anticuerpos/efectos de los fármacos , Reacción de Arthus/inmunología , Cobayas , Hipersensibilidad Tardía/inmunología , Inmunidad Celular/efectos de los fármacos , Inmunización , Inyecciones Intraperitoneales , Masculino , Ratones , Fagocitosis/efectos de los fármacos , Extractos Vegetales/farmacología , RatasRESUMEN
The polysaccharide fraction from Taraxii Herba showed potent immunopotentiating activities with antitumor activities. The fraction having small amount of protein inhibited the growth of solid tumor and increased peritoneal exudate cells and immunoorgan weights in normal mice, and also increased hypersensitivities in tumor bearing mice.
Asunto(s)
Adyuvantes Inmunológicos/farmacología , Antineoplásicos Fitogénicos/farmacología , Plantas Medicinales/química , Polisacáridos/farmacología , Adyuvantes Inmunológicos/aislamiento & purificación , Animales , Antineoplásicos Fitogénicos/aislamiento & purificación , Reacción de Arthus/inmunología , Ensayos de Selección de Medicamentos Antitumorales , Hipersensibilidad Tardía/inmunología , Masculino , Ratones , Ratones Endogámicos ICR , Tamaño de los Órganos/efectos de los fármacos , Tamaño de los Órganos/inmunología , Cavidad Peritoneal/citología , Hojas de la Planta/química , Polisacáridos/aislamiento & purificaciónRESUMEN
The effect of fish oil-derived omega-3 (omega-3) fatty acids on anaphylaxis, Arthus and delayed type hypersensitivity reactions in mice has been investigated. Mice on a normal chow diet were fed eicosapentaenoic acid and docosahexaenoic acid at a dose of 500 and 333 mg/kg/day, respectively, by a gastric tube over a period of 61 days. Control groups were given water, safflower oil or oleic acid. Anaphylactic and Arthus type reactions were induced in the mouse footpad using bovine serum albumin as an antigen. Carrageenin was utilized to produce a delayed type hypersensitivity reaction. The animals fed omega-3 fatty acids induced a more anaphylactic foodpad reaction. There was no significant effect of the diet on Arthus and delayed type hypersensitivity responses. There was no effect of the fish oil-supplemented diet on production of antibodies to bovine serum albumin. Synthesis of prostaglandin E2 by peritoneal macrophages was significantly inhibited in the animals fed omega-3 fatty acid-enriched fish oil, while leukotriene B4 production was not affected. These results suggest that a diet enriched in omega-3 fatty acids modulates production of arachidonic acid metabolites and this may influence anaphylaxis, but not Arthus and cellular mediated hypersensitivity responses.
Asunto(s)
Formación de Anticuerpos/efectos de los fármacos , Grasas Insaturadas en la Dieta/farmacología , Aceites de Pescado/farmacología , Inmunidad Celular/efectos de los fármacos , Anafilaxia/inmunología , Animales , Reacción de Arthus/inmunología , Dinoprostona/biosíntesis , Ácidos Docosahexaenoicos/farmacología , Ácido Eicosapentaenoico/farmacología , Hipersensibilidad Tardía/inmunología , Leucotrieno B4/biosíntesis , Macrófagos/efectos de los fármacos , Masculino , Ratones , Cavidad Peritoneal/citologíaRESUMEN
Methods are presented for an orderly search of a chemical file for complement inhibitors. Compounds are initially examined for intrinsic activity against dilute human components in vitro, using hemolytic assays to detect inhibitors of fluid phase C1, of late components lysis of EAC142, and of CVF-induced passive lysis of AET-treated human erythrocytes. Active compounds are then examined for activity against undiluted serum in vitro. Compounds passing this test are examined for activity in vivo against serum complement and complement-dependent lesions, viz. Forssman vasculitis, the reverse passive Arthus phenomenon, and Forssman shock. Methods are given for quantitation of these lesions.
Asunto(s)
Proteínas Inactivadoras de Complemento/aislamiento & purificación , Animales , Reacción de Arthus/inmunología , Proteínas Inactivadoras del Complemento 1/farmacología , Evaluación Preclínica de Medicamentos , Antígeno de Forssman/inmunología , Hemoglobinas/metabolismo , Masculino , RatonesRESUMEN
The development of novel anti-inflammatory drugs (AID) has been claimed to be dependent on the discovery of models of inflammation that differ from those currently used for drug screening, e.g. carrageenen paw oedema and u.v. erythema. We have thus evaluated the effect of a variety of drugs in a number of novel models of inflammation in the rat produced in the hind paw. We have utilized kaolin, zymosan, anti-rat IgG (anti-IgG) and the Reversed Passive Arthus (RPA) reaction to produce these oedema models. We found that the non-steroidal AID's, e.g. aspirin, flufenamic acid, indomethacin, naproxen, and phenylbutazone, were active in all four tests. Of the nine novel AID examined, levamisole and tetramisole demonstrated considerable activity in all four tests and dapsone was especially active in the anti-IgG and RPA tests. In contrast, the anti-rheumatic d-penicillamine was inactive in all four models. Each of the ten compounds tested which has been claimed to influence complement function, was active in the RPA but not in the kaolin model. These results are discussed in the context of the aetiology of each oedema and the suspected mode of action of the various drugs.
Asunto(s)
Antiinflamatorios/uso terapéutico , Modelos Animales de Enfermedad , Edema/tratamiento farmacológico , Animales , Antiinflamatorios/efectos adversos , Anticuerpos Antiidiotipos/farmacología , Reacción de Arthus/inmunología , Evaluación Preclínica de Medicamentos , Edema/inducido químicamente , Edema/inmunología , Inmunoglobulina G/inmunología , Caolín/farmacología , Masculino , Ovalbúmina/inmunología , Ratas , Úlcera Gástrica/inducido químicamente , Zimosan/farmacologíaRESUMEN
The effect of 4 immunosuppressive agents--hydrocortisone (HC), cyclophosphamide (CY), azathioprine (AZ) and methotrexate (MTX)--, on cutaneous delayed-type hypersensitivity (DTH) and Arthus reactions to the intradermal injection of ovalbumin (OA) in rats sensitized to OA in Freund's complete adjuvant (FCA) was studied. Multiple doses (daily for 4 days) were given either early, beginning on the day of sensitization, or late, beginning 9 days after sensitization. Single doses were given on the day of challenge with OA. All late multiple doses of drugs except HC depressed the DTH at 24 h in the following order of decreasing magnitude: MTX, CY, AZ. The DTH at 24 h was depressed by early multiple doses of MTX at all doses, by CY at all but the lowest dose, and by AZ at the intermediate dose; HC had no effect. When the drugs were given as single late doses, only CY at the lowest dose and MTX at the higher doses effectively depressed the DTH at 24 h. Increased Arthus reactions occurred after early and late multiple doses of HC and after a late single dose of CY at the highest dose. After late multiple doses the Arthus reaction was unaffected by either CY or MTX but was depressed by all doses of AZ. HC administered as 3 injections around the time of challenge markedly depressed the delayed and Arthus reactions. These results show that each of the 4 immunosuppressive drugs could depress DTH and Arthus reaction to OA, but the degree of depression varied with the time of drug administration relative to sensitization and challenge, and the dose of drug used. Histologic examination of skin test sites showed that an apparently negative reaction did not necessarily imply total absence of a cellular inflammatory response.
Asunto(s)
Reacción de Arthus/inmunología , Azatioprina/farmacología , Ciclofosfamida/farmacología , Hidrocortisona/farmacología , Hipersensibilidad Tardía/inmunología , Metotrexato/farmacología , Animales , Azatioprina/administración & dosificación , Ciclofosfamida/administración & dosificación , Relación Dosis-Respuesta a Droga , Femenino , Hidrocortisona/administración & dosificación , Leucovorina/farmacología , Masculino , Metotrexato/administración & dosificación , RatasRESUMEN
An egg albumin uveitis of the Arthus active type was developed in the rabbit. The experimental conditions were investigated in detail with regard to the following factors: influence of the number of sensitizing injections on serum antibody production, length of the recovery period which elapsed between sensitization and challenge, and influence of the size of the challenging dose on the severity of the inflammatory response. To develop the procedure optimally, emphasis was given to criteria of evaluation. Refractive index, protein and immunoglobulin assays in the aqueous humor of inflamed eyes were significantly correlated. These objective measures were considered more reliable than arbitrary grading systems. In addition, supportive histopathologic observations have been made in rabbit eyes. The above studies led to a reproducible model of uveitis in which drugs were tested topically. Dexamethasone phosphate in solution and indomethacin in suspension were effective in a dose-related manner. 6-Mercaptopurine did not demonstrate a useful anti-inflammatory effect.
Asunto(s)
Reacción de Arthus/inmunología , Modelos Animales de Enfermedad , Uveítis/inmunología , Administración Tópica , Animales , Anticuerpos/análisis , Humor Acuoso/análisis , Humor Acuoso/inmunología , Reacción de Arthus/tratamiento farmacológico , Reacción de Arthus/patología , Dexametasona/uso terapéutico , Relación Dosis-Respuesta a Droga , Evaluación Preclínica de Medicamentos , Proteínas del Ojo/análisis , Femenino , Inmunoglobulinas/análisis , Indometacina/uso terapéutico , Masculino , Mercaptopurina/uso terapéutico , Ovalbúmina/administración & dosificación , Ovalbúmina/inmunología , Conejos , Uveítis/tratamiento farmacológico , Uveítis/patologíaRESUMEN
The immune complex-induced reversed passive Arthus (RPA) reaction in the rabbit has been investigated as a screening test for detecting anti-inflammatory agents potentially more effective in the treatment of rheumatoid arthritis than those presently available. RPA lesions, characterized by edema, erythema, and hemorrhage, were elicited by intravenous injections of bovine serum albumin (BSA) followed by intradermal injections of rabbit anti-BSA antiserum. The anti-edema activities of compounds (mg quantities required for testing) were evaluated after their administration by the intradermal route (compounds admixed with antiserum) as well as by the intraperitoneal route. Of 14 reference anti-inflammatory agents tested by the intradermal screening procedure, only aurothioglucose and chloroquine were inactive. Other pharmacologically active compounds (e.g. antihistamines, anti-complement agents, cytotoxic-immunosuppressives) were also evaluated after their intradermal administration. Protoporphyrin, phloretin, and hexadimethrine bromide (Polybrene) were active. When whole antiserum, or the antibody fraction of the serum, was used to eliminate nonspecific edema, intraperitoneally administered reference agents were found to be effective in the RPA test.