Dose-response effects of TPI ASM8 in asthmatics after allergen.

BACKGROUND: TPI ASM8 contains two modified antisense oligonucleotides (AON) targeting the beta subunit (ß(c) ) of the IL-3, IL-5, GM-CSF receptors and the chemokine receptor CCR3. A previous study suggested that TPI ASM8 had broader effects than just inhibition of eosinophils in asthmatics. OBJECTIVE: We assessed whether TPI ASM8 caused a dose-dependent attenuation in the inflammatory and physiological changes after inhaled allergen challenge (AIC). METHODS: This single-center, open-label, stepwise-ascending dose study was conducted in fourteen stable, mild allergic asthmatics. Following placebo AIC, subjects underwent AIC after 4 days treatment with 1, 2, and 4 mg BID and finally 8 mg once daily (OD) of TPI ASM8, inhaled via the I-Neb™ nebuliser. Treatments were separated by 2-3-week washout periods. RESULTS: TPI ASM8 was safe and well tolerated at all doses. TPI ASM8 8 mg OD reduced eosinophils in sputum after AIC (by 60.9% at 7 h and 68.4% at 24 h post-AIC, P=0.016 and P=0.007, respectively). Additionally, TPI ASM8 8 mg OD significantly attenuated the early and late airway responses as shown by the reduction in the area under the curve by 45% (P=0.016) and 59%, (P=0.0015), respectively, the increase in eosinophil cationic protein (ECP) by up to 57% (P=0.021), and airway responsiveness to methacholine by more than 1 doubling dose (P=0.012). A dose-response relationship was noted, and efficacy was maintained with once per day administration. CONCLUSIONS: TPI ASM8 attenuated a broad range of inflammatory and physiological changes after AIC, suggesting that CCR3, IL-3, and GM-CSF also are important targets for the management of asthma.

Evaluation of cortisone-heparin and cortisone-maltose tetrapalmitate therapies against rodent tumors. I. Biological studies.

The antitumor activity of either cortisone-heparin or cortisone-maltose tetrapalmitate combination or both was tested against two animal tumor models. The first model was orthotopically implanted bladder tumor established in syngeneic Fisher 344 rats. Shrinkage and growth arrest of the tumors were induced by cortisone and amplified by its combination with either heparin or maltose tetrapalmitate (MTP). The second model was trocar implanted C3HBA mammary tumor piece s.c. in syngeneic LPS and MTP responder C3H/HeN and non responder C3H/HeJ mice. The tumor was sensitive to growth inhibition by cortisone-MTP in the C3H/HeN but not by cortisone alone or cortisone-heparin. Tumor implanted in C3H/HeJ was much less sensitive to cortisone-MTP. Cortisone could be replaced by 17-alpha-hydroxyprogesterone, but not by cortexolone.

Evaluation of cortisone-heparin and cortisone-maltose tetrapalmitate therapies against rodent tumors. II. Pathological studies.

Pathological studies were undertaken in three tumor-host models which were subjected to cortisone based treatments. The first model was Fisher 344 rats with established orthotopically implanted syngeneic bladder tumor. Cortisone-herapin and cortisone-maltose tetrapalmitate (MTP) treatments induced focal areas of tumor necrosis and necrobiosis, whereas cortisone alone caused necrobiosis. The second model was C3HBA mammary tumor fragments implanted subcutaneously in syngeneic MTP responder C3H/HeN and MTP non-responder C3H/HeJ female mice. Only cortisone-MTP treatment led to an absence of capillary extension from surrounding blood vessels into the scant tumor stroma. The third model, ethyl carbamate induced primary lung cancer in AJ mice, was tested only with cortisone-herapin combination. The treatment caused central zones of necrosis.

Antitumor efficacies of maltose tetrapalmitate immunotherapy alone and in combinations with radiotherapy and with cyclophosphamide chemotherapy against dimethylhydrazine induced colon and anal cancers in CDI mice.

Treatment of human colonic cancer in early stages when the process is still limited to the colonic wall is primarily surgery. We wished to see if maltose tetrapalmitate (MTP) immunotherapy alone or in combination with radiotherapy (R) and cyclophosphamide (C) chemotherapy would be effective against primary colon cancer in a fashion similar to that reported by us for primary liver cancer (Anticancer Research 6: 245-250, 1986). One hundred female CD1 mice were subjected to dimethylhydrazine (DMH) treatment once a week for 26 weeks, a period one week before which, colon cancer was histologically documented in each animal of a group that was sacrificed. Surprisingly, many of the animals harboured early anal cancer as well. At 28 weeks, 85 of the available animals were divided into 6 groups that received: Gr. 1, no treatment; Gr. 2, MTP alone (M); Gr. 3, radiotherapy alone (R); Gr. 4, cyclosphophamide alone (C); Gr. 5, R + C; Gr. 6, M + R + C. Criteria of treatment efficacy were: number, size and staging of colorectal tumors and the incidence and the size of anal tumors at death. Mean survival time was also determined although it remained a questionable criterium since most animals died due to complication (hepatic toxicity, pyelonephritis, thrombose) elicited by DMH, R and C toxicities and not as a result of colonic tumor size or metastases. As a single therapy, M appeared to be superior to either R or C alone. However, R + C combination was effective and was further improved upon by its association with M. With the triple combination, (M + R + C), lesions of both cancers decreased in size and/or number and the colon cancer histologically eclipsed from 46% of the treated animals.

Effects of single and combined maltose tetrapalmitate immunotherapy, cyclophosphamide chemotherapy and radiotherapy on ethyl carbamate accelerated primary lung cancer in A/J mice.

A/J mice were given ethyl carbamate to accelerate and to raise to 100 percent the incidence of lung tumours at 34 weeks (day 237) of age. The animals were then divided into groups which received the following treatments: group 1, no treatment; group 2, MTP alone; group 3, radiotherapy alone; group 4, cyclophosphamide alone; group 5, radiotherapy + MTP; group 6, MTP + cyclophosphamide; group 7, radiotherapy followed by cyclophosphamide and group 8, MTP and radiotherapy together followed by MTP and cyclophosphamide. Except for radiotherapy, which was given for 5 consecutive days, MTP and cyclophosphamide were continued till the death of the animals. The treatment efficacies were evaluated by the number and size of tumour nodules, taking into consideration the survival time of the animal. Animals in groups receiving cyclophosphamide died earlier (between days 290 and 315) due to its toxic effects, and half of the radiotherapy-MTP were sacrificed at day 314 for comparison. Although cyclophosphamide alone and radiotherapy plus cyclophosphamide demonstrated antitumour activity, the number of tumour nodules and the nodule diameter were reduced most effectively in group 8 (receiving MTP, radiotherapy and cyclophosphamide). Among the animals in the non-cyclophosphamide group, radiotherapy alone was ineffective. MTP given before and after radiotherapy (group 5) kept tumour volume in control although this group died suddenly. The animals receiving only MTP died between day 430 and 470. The number of tumour nodules and the nodule diameter in the MTP group were, however, significantly reduced when compared to controls or radiotherapy group animals dying at or near the same time.

Combined radiotherapy, chemotherapy, and maltose tetrapalmitate immunotherapy in the treatment of 4'dimethylaminoazobenzene-induced liver cancer.

Therapeutic effects of radiotherapy (R), chemotherapy, and maltose tetrapalmitate (MTP) immunotherapy alone and in combinations were tried against 4' dimethylaminoazobenzene (DAB) induced primary liver cancer in Wistar rats in three separate protocols. Rats were fed a low protein synthetic diet containing 0.06% DAB for 90-120 days. Around 90 days, liver cancers developed in all the animals. In the first protocol, animals were either left untreated or treated with cyclophosphamide (Cy), MTP (i.p. or oral) and Cy plus oral MTP. Rats in the MTP (i.p.) group maintained a steady liver weight but neither Cy nor Cy + MTP influenced the survival time or liver weight. In the second protocol, R as well as a 3-drug combination at 2 dose levels were tried alone and with MTP before or soon after cessation of DAB feeding. Survival times were decreased by R and chemotherapy due to combined toxicities of DAB and treatments and were partially restored by MTP. In the third protocol, MTP, R, and Cy were each tried alone and in combinations, 21 days after cessation of 100-day DAB feeding. Increase in survivals were obtained by each treatment, although tumor weight was best controlled by triple R+ Cy + MTP combination.

Effect of membrane fatty acid composition on radiosensitivity of V79 Chinese hamster cells.

Exponentially growing V79-379A Chinese hamster fibroblasts were transferred to low-lipid medium enriched with a single fatty acid of the C18 series. After 24 h at 37 degrees C, the fatty acid composition was determined by gas chromatography of the corresponding methyl esters for the total lipid extracts of the cells and for the nuclear membrane fraction. Radiation survival curves, based upon a clonogenic assay, were obtained by irradiation with low dose-rate 60Co gamma rays at either 4 degrees C or room temperature. We observe no effect of fatty acid upon radiosensitivity of these cells at either temperature, in confirmation of published reports with other mammalian cell lines.

Combination of 60Co gamma-radiation, misonidazole, and maltose tetrapalmitate in the treatment of Dunning prostatic tumor in the rat.

Maltose tetrapalmitate (MTP), a synthetic nontoxic immunoadjuvant, the radiosensitizer misonidazole (MISO), and 60Co gamma-radiation, alone or in combination, were used in the management of Dunning prostatic tumor in the rat. Nine groups of 10 rats each were used to assess the efficacy of various therapeutic modalities. Tumor growth rates and animal survival times were determined for each group. Radiation was more effective when combined with MTP, but the adjuvant must be present when radiation is given for synergism to occur. MISO was as effective as MTP when used with radiation, but combining them cancels out their individual effects. In a clinical situation it would be advantageous to use separately the synergisms existing between MISO and radiation on the one hand and MTP and radiation on the other hand.

Systemic, bladder wall, and bladder tumor concentration of misonidazole following intravesical administration in the rat.

The hypoxic-cell radiosensitizer misonidazole was administered intravesically to normal and to bladder tumor-bearing female Fischer rats. Drug concentration was measured in the bladder wall, the tumor and in the serum using high pressure liquid chromatography at different times following administration. The data shows that misonidazole is readily adsorbed by the bladder wall and the tumor with tissue levels reaching up to 10 times those measured in the serum.

Dietary immunostimulation: interaction with BCG and LPS.

The action of BCG and LPS mR595 used in conjunction with a formula-defined diet is dependent on the administration timing and resembles that of interacting adjuvants affecting different elements of the immune system.

The role of diet on 5-fluorouracil toxicity.

In three experiments performed at two month intervals, rats eating a nutritionally adequate defined-formula diet (DFD) exhibit a remarkable similarity in the response to 5-FU as measured by survival time, leukopenia and weight loss. Conversely, the corresponding control groups eating the standard Purina rat food show a different degree of tolerance to 5-FU, which varies from a level similar to that of the DFD groups to twice that value. It is suggested that certain protective factors against 5-FU toxicity present in natural food are absent in formula diets produced on the basis of standard requirements for normal growth and nutrition. The present studies indicate that such protective factors are neither vegetable fibers nor arachidonic acid. The importance of food intake in experimental toxicology and clinical pharmacology is emphasized.

Dietary enhancement of intestinal radioresistance during fractionated irradiation.

Rats fed laboratory chow or elemental diet 3 were given fractions of 240 rads of 60Co gamma-radiation abdominally (1200 rads/week) until all animals had died. Changes in appetite, body weight, and mortality were monitored as a function of the cumulative dose received. More radiation was needed in the diet-fed group to achieve both 0 and 100% mortality, a difference of 37% at the mean lethal dose level. Both groups developed similar progressive anorexia but the diet-fed animals lost weight more slowly. Data indicate that basic intestinal radio-resistance is enhanced by feeding the elemental diet.

Enhanced 5-F.U. mortality in rats eating definded formula diets.

Rats eating nutritionally adequate defined-formula diets (D.F.D) before and following lethal i.p. injection of 5-fluorouracil exhibit increased mortality and shorter survival time than rats eating conventional Purina rat chow. Neither the presence of proteins in hydrolyzed form nor the nitrogen level in the D.F.D appear to be the crucial factor of enhanced lethality. The possible mechanisms and implications of the phenomenon are discussed.

Immunostimulation by a formula-defined diet.

A formula-defined diet (diet 3, table 1) acted as an adjuvant as an adjuvant in the response of the immune system to SRBC in the rat. Similar minimum and maximum antibody levels were measured in both males and females. In the males, the maximum was reached with diet 3 alone while females required the complementary action of diet 3 and LPS mR 595.

Effects of feeding an elemental diet during abdominal irradiation.

Rats fed with normal laboratory feed or elemental diet 3200-BD (Mead Johnson) were administered fractions of 240 rd (-60Co gamma-radiation) abdominally. Consumption of food and its nutrition efficiency were the parameters studied. Anorexia occurred rapidly following only one fraction of 240 rd and was unaffected by the feeding of the diet. Animals fed the latter, however, recovered normal appetite much faster upon completion of abdominal irradiation than those fed normally. While nutritional efficiency of the diet was smaller than that of normal feed with the controls, the opposite was obtained with irradiated animals.