MXPA97009756A - The use of a prolactin reducer to prepare useful containers to inhibit neoplastic diseases in mamife - Google Patents

Abstract

This invention relates to the use of a porlactin reducer for preparing compositions useful for inhibiting the growth of neoplasms in a mammal having a prolactin profile, the method involves comparing the prolactin profile of the affected mammal with a normal profile of prolactin for healthy mammals. of the same species and sex and adjust the prolazine profile of the affected mammal to adjust or approximate the normal prolactin profile of a mammal of the same species and sex as the affected mammal, thereby inhibiting neoplastic growth.

Description

THE USE OF A PRQLACTINE REDUCER TO PREPARE USEFUL COMPOSITIONS TO INHIBIT NEOPLASTIC DISEASE IN MAMMALS BACKGROUND OF THE INVENTION FIELD OF THE INVENTION This invention relates to methods for inhibiting neoplasms and their metastases. More particularly, this invention relates to methods that employ the alteration of circadian prolactin rhythms to inhibit or excise neoplasms and their metastases.

Prolactin and circadian rhythms Research has shown that circadian rhythms play important roles in the regulation of prolactin activities and vice versa. Publications such as Meier, fl.H », Gen. Cornp. Endocrinol 3 (Sup 1): 488-508. 1972; Meier, fi.H .. Trans. Rrn. Fish. Soc. 113: 422-431, 1984; Meier, A.H. , and others, Current Ornithology II (ed Johnston R.E.) 303-343, 1984; Cincotta, A.H. and others, 3. Endocrinol. 120: 385-391, 1989; Meier, A.H., A er. Zool. 15: 905-915, 1975; Meier, A.H. , Hormonal Correlates of Behavior (eds. Eleftherton and Bprott) 469-549, 1975 illustrate how circadian rhythms regulate prolactin activities.

The daily variations rosultan-.es in response of several cell types to the prol.Ana play a major role in the regulation of numerous physiological procedures, including fat storage, lipogenic response to insulin, migratory behavior, crop development of pigeon and breast development (Meier, AH, Gen. Cornp. Endocrine! 3 (Suppl 1): 488-508, 1972; Meer, AH, Ainer. Zool. 15: 905-916, 1975; Meier, AH and others Science 173: 1240-1242, 1971). In the regulation of one of the previous physiological activities, it can be observed that prolactin produces a stimulating or an inhibitory effect on a given activity, or has no effect on it. These variable effects have recently been noted that in animals they are a function of the time of the endogenous daily peak (ie, acrophase) of the prolactin concentration rate in the plasma or a function of the exogenous hormone (or a substance that increases prolactin levels) or the relationship between the endogenous p and any induced peak. In addition, high levels of μlactoma restricted to a discrete daily interval have a much greater physiological effect (eg, etabolic) in animals than constant high levels during a day (Cmcotta, AH and other Horrn. Metab. Res. : 64-68, 1989; Borer, KT m The Hamster: Reproduction and Behavior (ed. Siegel, HI) 363-408, 1985). These findings demonstrate the existence of daily response rhythms to the prolactum by certain cell types.

The first demonstration of a daily variation in the physiological response to any hormone was the drastic variation in response of the fattening to prolactin in the white-necked sparrow (Meier, AH and others Gen. Comp.Endocrinol., 8: 110-114, 1967). Mid-day injections of a 16-hour daily photoperiod stimulated 3-fold increases in body fat levels, while injections given early in the photoperiod reduced fat reserves by 50%. Said daily variations in prolactin fattening responses were subsequently demonstrated in numerous species of all major vertebrate classes (Meier, AH, A. Zool, 15-905-916, 1975, Meier, AH, Hormonal Correlates of Behavior. eds, Eleftherton and Sprott) 469-549, 1975) indicating the fundamental nature of such a temporary organization. The rate of fattening response persists under constant light conditions (Meier, AH, et al. Proc. Soc. Exp. Biol. Med. 137: 408-415, 1971) indicating that, like many other endogenous rare variations, it is a circadian rhythm. Additional studies have shown that circadian rhythms have major roles in regulating numerous physiological activities, such as lipid metabolism and body fat stores (Meier, AH et al., Current Ornithology II (ed Johnston RE) 303 -343, 1984; Meier, A.H. Amer. Zool. 15: 905-916, 1975; Meier, A.H., Hormonal Correlate of Behavior (eds. Eleftherton and Sprott) 469-549, 1975; Meier, A.H. and others 3. rn. Zool. 16-649-659, 1976); Cincotta et al., Life Sciences 45: 2247-2254, 1989; Cincotta and others, Ann. Mutr. Metab. 33: 305-14, 1989; and Cincotta and others Horrn. Metabol. Res. 21: 64-68, 1989. These experiments showed that an interaction of circadian rhythms of liporegulatory hormones (stimuli) and circadian responses to these hormones (in target cells) determines the amount of lipogenesis and fat reserves. Therefore, elevated plasma prolactin concentrations (which serve as the stimulus) appear during the daily interval of responses of maximum fattening to prolactin in fat animals, but appear at other times of day response in lean animals (Meier, AH, Arner, Zool, 15: 905-916, 1975, Meier, AH, Hormonal Correlates of Behavior (eds. Eleftherton and Sprott) 469-549, 1975; Speiler, RE and others Nature 271: 469-151 471, 1978). Similarly, plasma insulin levels (acting as a stimulus) are higher during the daily interval of greater hepatic lipogenic response to insulin in obese hamsters, but at a different time of day in thin hamsters (deSouza , CJ et al., Chronobiol.

Int. 4: 141-151, 1987; Cincotta, A.H. and others, O.Endocr. 103: 141-146, 1984). The phase relationships of these stimuli and response rhythms are believed to be expressions of neural circadian centers which in turn can be reinitialized by neurotransmitter agents and hormone injections ? r. (including prolactin) to produce either fat or thin animals (Meier, A.H., Trans.An.Finn. Soc. 113: 422-431, 1984; Meier, A.H. and others, Current Ornithology II (ed Dohnston R.E.) 303-343, 1984; Cincotta, A.H. and others, 3. Endocrinol 120: 385-391, 1989; Ernata, A.C. and others, 3. Exp. Zoo !. 233: 29-34, 1985; Cincotta, A.H. and others, Chronobiol, Int'l 10: 244-258, 1993; Miller, L.3. and others, 3.1nterdisc. Cycles Res. 14: 85-94, 1983). Accordingly, the administration or increase of programmed prolactin has been shown to act directly on tissues (eg, liver in lipogenesis) that undergo circadian rhythms of response to the hormone to produce immediate variations in net physiological effects (Cincotta, AH, et al. , Horrn, Metab. Res. 21: 64-68, 1989) and also acts indirectly reinitializing one of the circadian neuroendocrine oscillations of a multidisciplinary circadian pacemaker system to establish different phase relationships between multiple circadian expressions (neural, hormonal and of tissue) that control lipid metabolism (Meier, AH, Trans.Ar.Fish.Soc. 113: 422-431, 1984; Meier, AH et al., Current Ornithology II (ed. Johnston RE) 303-343, 1984; Cincotta, AH and others, 3. Endocrinol, 120: 385-391, 1989, Ernata, AC et al, 3. Exp. Zool, 233: 29-34, 1985, Cincotta, AH et al., Chronobiol, Int'l 10 : 244-258, 1993, Miller, L.3 and others, 3. I interdisc Cycles Res. 14: 85-94, 1983). The inventors of the present have shown above that prolactin, or substances that affect circulating prolactin levels, also affect circadian rhythms and in fact can be used to modify these rhythms (so they closely resemble the rhythms of thin individuals). , healthy, young people of the same sex) and reinforce these rhythms (so that the modified rhythms persist in the modified condition). See, e.g., patent applications of E.U.A. 08 / 158,153, 07 / 995,292, 07 / 719,745, 07 / 999,685, 08 / 171,569, and patent of E.U.A. No. 5,344,832. This previous work by the inventors of this has been clinically proven in humans affected by various physiological disorders (obesity, diabetes, atherosclerosis, hypertension, immune dysfunction and others) with good results. In particular, in the patent application of E.U.A. No. 07 / 995,292 and in its continuation in part series No. 08 / 264,558, filed on June 23, 1994, the present inventors describe a method for the reduction in a subject, vertebrate animal or human being, of reserves of fat in the body, and reduction of at least one of insulin resistance, hyperpulsulinernia and hyperglycemia, and other metabolic diseases, especially those associated with diabetes of type II. Very specifically, the above application describes methods to: (i) evaluate the daily prolactin level cycles of a normal vertebrate human or animal (healthy) (without obesity, disease or other disorder); (n) diagnose aberrant cycles of daily prolactum level of a vertebrate human or animal; (m) determine the appropriate adjustments that need to be made to normalize such aberrant cycles of prolactin level; This method involves the administration of at least one of a prolactin reducer and / or a prolactin enhancer in a predetermined first time (or times) within a 24 hour period (if only one prolactin reducer is administered) and / or in a predetermined second time (or times) of a 24-hour period (if a prolactin enhancer is administered). This therapy, when continued for several days, weeks, or months, results in long-term adjustment of aberrant or normal prolactin level cycles so that they conform (or approximate) to normal prolactin level cycles. In most cases, this benefit persists in the long term even after cessation of therapy. As a result, aberrant physiological parameters associated with several metabolic disorders are stored at normal levels or modified to reach normal levels. Although this method is applied to all people who have aberrant prolactin levels at least for a portion of a 24-hour period, importantly, there is no teaching of the possibility of applying it to people with neoplastic disease, nor is teaching possibility of applying this method to the treatment of neoplastic conditions.

CORTICOSTERONE AND CIRCADIUM RHYTHMS The secretion rates of corticosteroid in humans are high early in the morning but low early in the night. Plasma levels in the plasma range from as high as 0.2 rncg / rnl to one hour before waking in the morning and as low as around 0.05 rncg / ml at approximately 12 AM. This effect is the result of a cyclic alteration of 24 hours in the signals of the hypothalamus that produce corticosterone secretion. When a mammal changes sleep habits, the cycle changes accordingly. On the contrary, when the cycle changes, sleep habits also change. Therefore, the administration of corticost rona can be used to synchronize the circadian rhythms of a number of experimental mammals that have been deprived of a photoperiod by exposure to constant light, as is done in some of the more described examples. ahead. The pattern of corticosterone secretion is different for each species but can be easily determined by testing the hormone at various time intervals during portions of dark and light from the photoperiod. Although it was well known in the art that it was possible to control many rnetabolic disorders by adjusting prolactin rhythms, it was completely surprising and unexpected to find whether the prolactin rhythms in mammals affected by neoplasm and metastases were adjusted to conform to or achieve the rhythms found in young individuals. , healthy, thin the same species and sex, neoplastic growth and etastat co was inhibited to a very important degree ..

BRIEF DESCRIPTION OF THE INVENTION It has long been known that mammals (including humans) suffering from neoplastic diseases have abnormal prolactomy profiles. Now it has unexpectedly been discovered that neoplasms and their metastases in mammals (including humans) can be treated by modifying the abnormal prolactin profile of the affected mammal with neoplastic disease, for which the profile approximates or conforms to the prolactin profile of a slender, young, healthy mammal of the same species and sex. The profile of abnormal prolactin of the affected animal can be modified by i) direct administration of prolactin, n) adjustment of the prolactin profile by timely administration of prolactin modulators, ie prolactin enhancers and / or reducers, on ) the reimmediaization of the circadian rhythm of the affected mammal to a normal phase and amplitude through the timely administration of prolactin enhancers (such as rnelatonma) and prolactin reducers (such as rornoc p ina). Therefore, one aspect of the present invention is a method for treating or inhibiting neoplasm and its metastases in mammals by administering to the mammal a prolactin reductant and / or enhancer or sequential administration! of a prolactum increment and reducer in a predetermined time or period during a 24-hour period that results in the modification of the mammal's abnormal prolactum profile so that it approximates or conforms to the prolactum profile of a young healthy mammalian rnarn of the same species and sex. Another aspect of the present invention is directed to a method for treating or inhibiting neoplasm and its methastases on a long-term basis by continuing the earlier () timely administration of prolactin reducer and / or prolactin enhancer until the prolactin The subject's condition is reinitialized and persists in this reinitialized condition for a prolonged period even after cessation of therapy, resulting in persistent neoplastic growth inhibition. Therefore, the present invention is directed to the treatment or inhibition of growth of neoplasms in mammals by adjusting the circadian rhythm of prolactin. The method of the invention achieves the inhibition of neoplastic growth by normalizing the circadian rhythm for prolact na of the sujo. or receiving treatment to resemble that of a healthy young subject. The advantages of the present invention include. the ability to fight neoplasms without the debilitating effects of surgical agents. the ability to inhibit the metastatic growth of neoplasms that often accompany removal of the primary neoplastic tissue. the benefits of inhibition and neoplastic growth treatment of the present invention may persist in the long term and even after the administration of prolactum modulators has been discontinued. Other aspects and advantages of the present invention will be apparent from the following description taken together with the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 illustrates the profile of normal or baseline p olicy for male and female humans. Figure 2 is the rhythm curve or prolactum daily profile for mice with or without an implanted EMT-6 tumor. Figure 3 is the rhythm curve or prolactum daily profile for cancer patients with tumors. Figure 4 is a bar graph illustrating the effect of timely prolactin injections on EMC-6 tumor growth in Balb / C mice whose circadian rhythms are being fixed by corticosteroin injections.

DETAILED DESCRIPTION OF THE INVENTION All patents, patent applications and literature references that are described in this specification are incorporated herein by reference. In the case of a conflict in terms of terminology, the present description includes its definition controls. "Prolactin reducer" refers to a substance or composition that has the ability to reduce circulating prolactin levels under administration to a mammal; "prolactin enhancer" refers to a substance or composition that has the ability to raise circulating prolactin levels, and includes prolactin rnisrna. Prolactin reducers and prolactin enhancers are collectively known as "prolactin modulators." "Prolactin Profile" of a subject is an illustration of circulating prolactin levels and their variation over all or a part of a 24-hour period, and therefore is an expression of all or part of the daily prolactin rhythm in the plasma of the subject.

"Sano" is a young, thin subject without disease and including malignancies, dysfunctions of the immune system and metabolic abnormalities. A healthy subject is one with a normal prolacton profile, that is, a prolactm profile that does not deviate from the baseline of the species and sex of the subject in more than one standard error of the mean (SEM) . The normal or baseline profile of prolactin for healthy male and female humans is illustrated in Figure 1. To avoid "false positives", a subject will not generally be considered to have an abnormal prolactin profile at least. a) the level of prolactm in the blood during the subject's day is at least one SEM higher than the baseline at two (or more) points in time during the period of the day separated by more than one and preferably by at least two hours; or b) the level of prolactin in the blood at the time of the subject's day is at least 2 SEM higher than the baseline at a point in time during the time of day; or c) the prolactum level in the blood during the subject's night time is at least 1 SEM below the baseline at two (or more) points in the separated time (as in (a)); or d) the prolactum level in the blood during the subject's night time is at least 2 SEM below the baseline at a point in the time during the night time. The lines b prolactin salts for male and female humans are illustrated in figure 1. An SEM during waking hours (07:00 - 22:00) is approximately 1-2 ng / ml for people of sex male and of approximately 1-3 ng / rnl for female persons; an SEM during the night time (22:00 - 07:00) is approximately 3 ng / rnl for males and approximately 3-6 ng / ml for females. The characteristics of the rhythm or daily profile of the level of prolactin that must be reached or conformed in humans includes achieving low levels of prolactin (2-7 ng / rnl of plasma) for males and 2-10 ng / rnl for people female) during most or all of the period between 07:00 and 22:00 h. Ideally, a peak prolactin level should also be achieved between the hours 22:00 and 07:00 (preferably between 1:00 and 4:00) (the peak should be at least 10 ng / ml and preferably between 10-15 ng / ml for males and at least 15 ng / ml and preferably between 15 and 25 ng / ml for females).

Effects of prolactin modulators on neoplastic disease The present invention provides a method for treating and inhibiting the growth of neoplasms and their metastases (e.g., reducing the amount of neoplastic tissue, or reducing the netastatic load after primary tumor removal). , if the neoplasm is solid) in mammals with a substantial neoplastic tissue load or with potential metastatic growth after removal of a primary neoplastic tissue mass. This can be achieved by administering a prolactin modulator at predetermined times for a period of 24 hours. The time for administration of the prolactin modulator is selected to adjust the prolactin profile of the mammal receiving the treatment to conform to or approximate the prolactin profile of a healthy mammal of the same sex and species. It has been found that the administration of prolactin enhancers is inhibitory to neoplastic growth in mammals when given at scheduled intervals over a period of 24 hours corresponding to the peak secretion of prolactin in healthy mammals. Prolactin injections programmed into mice that have neoplasm that have had their circadian rhythms synchronized with either a defined photoperiod or with corticosterone injections were shown to exhibit a reduced neoplastic tissue load compared to mice that had neoplasm that did not receive injections of prolactin programmed. It has also been found that the effect of modulating prolactin in vivo of neoplastic tissue inhibitory responses in vivo and metastasis is dependent on the time of day.

The dependent role of the time of day for prolactin in the inhibition of neoplastic disease is also indicated by results of experiments in mice, which reduces the levels of prolactin in the blood (by administration of a prolactin reducer) during specific daily intervals of lack of inhibitory response of neoplastic growth to exogenous prolactin. Time-course response studies with bromocriptine, a dopamine D2 agonist that inhibits endogenous prolactin secretion indicates that bromocriptine increases the inhibition of neoplastic and metastatic growth when administered at predetermined times over a 24-hour period corresponding to the nadir secretion of prolactin in healthy animals. These results are illustrated in example 5. Further confirmation of the time-dependent role for prolactin in the inhibition of neoplastic growth is illustrated in example 6. In this experiment, prolactin levels in the blood of mice are reduced by the administration of bromocriptine, a prolactin reducer, during the specific daily interval of lack of response of neoplastic growth inhibitory activity to prolactin as found in example 5 above and prolactin levels are increased by the administration of melatonin , a prolactin enhancer to determine the specific daily interval of increased response of neoplastic growth inhibitory activity to prolactin. It is found that the combination of prolactin enhancer administration in the. time during a 24-hour period when prolactin levels reach a peak in healthy mice and the administration of prolactin reducer in the period during a 24-hour period when prolactin levels are at their nadir exert a potent inhibitory effect on the growth of neoplasm. The above results indicate the inhibitory effects of neoplastic growth of prolactin levels and the relationship between the inhibition of neoplastic growth to exogenous prolactin (or prolactin enhancers or reducers) and the time of day of reduction or increase of prolactin. Although in the previous experiments they are conducted in mice, depend on aspects of physiology that are common for mammals that have a daily prolactin rhythm including humans. These results show that prolactin levels in the blood can be manipulated during predetermined intervals to obtain a desirable result with respect to the inhibition of growth of neoplasms and their metastasis. In accordance with the method of the present invention, the alteration of prolactin levels of a subject at particular hours of the day provides methods for inhibiting neoplastic growth in the subject or inhibiting the growth of metastases in a subject. The method can be used in all types of neoplasm, including but not limited to sarcomas, carcinomas, gliobastornas, norn rnel, lympholas, adenomas and leukemias.

USE OF PROLACTIN MODULATORS TO INHIBIT NEOPLASMS AND THEIR METASTASIS ADJUSTMENT OF PROLASTIN RHYTHMS OF SUBJECTS WITH NEOPLASMS AND / OR MET STASIS It is known that healthy adult young mammals of a given species (and given sex) eg, humans (not suffering from hormonal or metabolic disorders or cancer) have a highly predictable daily prolactin rhythm or profile. The baseline curve for healthy male and female humans in Figure 1 is derived from said healthy young individuals. The phase relationship between daily bitrno peaks and response (inhibition of neoplastic growth) to the stimulus (prolactin in the plasma) to prolactin has been found to be important in the neoplastic growth inhibitory activity. Environmental and pharmaceutical factors that influence any of these rhythms can be expected to have an impact on neoplastic growth. Humans with a neoplastic disease, such as breast cancer, have disturbed prolactin rhythms, which is evident in comparing the prolactin rhythms of healthy women with the rhythms of women with breast cancer. shown in Figures 1 and 3, respectively. Humans with neoplastic disease can therefore benefit to a significant degree by adjusting their daily prolactin rhythms (as expressed by their prolactin profile) to conform to or approximate the normal prolactin curve or basal line of Figure 1. An adjusted prolactin profile approximates a normal or healthy profile if all or part of the abnormal profile moves in the correct direction at least 2 ng / ml. Before this adjustment can be achieved: i) the prolactin levels of the person who has neoplasm should be evaluated by testing blood samples from the person who has neoplasm at certain separate intervals within a 24-hour period and ii) the resulting prolactin profile of the person who has neoplasm should be compared to the prolactin profile for a healthy person of the same sex. Depending on the difference between i) and ii), the adjustment then involves administering one or both of the following: a) a prolactin reducer in a predetermined first time (or in more than a predetermined first time) and in a first effective amount to reduce prolactin levels at the time of day if these levels are too high; and b) a prolactin enhancer in a predetermined second time (or in a plurality of seconds predetermined times) and in a second effective amount to increase prolactin levels of night time if these levels are too low. In general, if a substance that alters the level of prolactin is to be administered, the appropriate administration time should be left to allow the substance (depending on its pharmacokinetic properties) to affect the prolactin levels in such a way that the levels of prolactin are modified during the time of the appropriate day. Thus, the substance that causes prolactin will be administered in the following manner: (a) if prolactin is administered, it will be administered, preferably by injection, during the time interval when prolactin levels need to be elevated; (b) if a prolactin enhancer different from prolactin is administered, it will be administered during or sometime shortly before the time interval when prolactin levels need to be elevated (how much time before, depends on pharmacokinetic properties: it has been generally found that 0 to 3 hours before, it is effective); and (c) if a prolactin reducer is administered, it will also be administered during or shortly before the time when prolactin levels need to be reduced (again, it has been generally found that from 0 to 3 hours before, it is effective). In the method of the present invention, "prolactin enhancer" includes prolactin as well as substances that increase circulating prolactin levels (eg, by stimulating the secretion of the prolactin itself). Non-limiting examples of a prolactin enhancer include prolactin; rnelatonin; dopamine antagonists such as metoclopramide, haloperidol, pyrnozide, phenothiazine, domperidone, sulpiride and chlorprornazine; serotonin agonists, ie, MAO-A inhibitors, for example, pargyline, synthetic analogs of morphine, eg, rnetadone; antiemetics, for example, metoclopramide; estrogen; and various other serotonin agonists, for example, tryptophan, 5-hydroxytryptophan (5-HTP), fluoxetine, and fenflurarynin. In addition, the non-toxic salts of the above prolactin enhancing compounds formed from pharmaceutically acceptable acids are also useful in the practice of this invention. It has been found that rnelatonin and 5-HTP are particularly useful in the practice of this invention. Non-limiting examples of prolactin reducers include prolactin-inhibiting dopamine agonists (D2 agonists), such as dopamine and certain prolactin-inhibiting compounds related to ergot. Non-limiting examples of dopamine agonists are 2-rhino-alpha-ergocriptine; 6-Rethyl-8-beta-carbobenzyloxy-arninoethyl-10-alpha-ergoline; 8-acylarninoergolines are 6-rnetyl-8-alpha- (N-acyl) arnino-9-ergoline and 6-rnetyl-8-alpha- (N-phenylacety-D-arnino-g-ergolma; ergocormna; 9,10-dihydroergocornin; and ergolinas; D-2-halogeno-6-alkyl-8-substituted, for example, D-2-brorno-6-rnet? L ~ 8-cyanorneti ergoline, carb-dopa and L-dopa, and lisup a. The non-toxicity of the μ-lactin-reducing compounds formed with pharmaceutically acceptable acids are also useful in the practice of this invention. It has been found that bro-scripphin, or 2-bruno-alpha-ergocptin, is particularly useful in the practice of this It is expected that the modulation of the inhibition of neoplastic growth induced by prolactin inhibitors or reducers will be dependent on the dose on a dose scale.In general, in the treatment of mammals, the doses of the reductant and / or Increased! - of prolactm, respectively, are each given generally once a day, usually during a period of time It lasts approximately one year approximately one year, but the treatment can continue indefinitely (if necessary or desired) for months or even several years. The preferred prolactone reducer (accelerated release brornocpptine) is administered at daily dose levels ranging from about 3 micrograins to approximately 300 initiation, preferably from about 10 micrograms to about 100 micrograms per kilogram of protein. body weight, and a preferred prolactone metabolite, the rnelatomine, is administered at daily dose levels ranging from about 10 inierograms to about 800 micrograms, preferably from about 10 micro-grams to about 200 micro-grams, per kilogram of body weight per day to modify, or alter, the prolactin profile. Another preferred prolactin enhancer, 5-hydroxytryptophan, is administered at daily dose levels ranging from about 500 micrograms to about 13 milligrams per kilogram of body weight, preferably from 1 milligram to 2.5 milligrams per kilogram of body weight. The exact dose within these scales that will be administered to each subject will depend on the particular prolactin modulator, the age of the subject, the stage of the disease, the physical condition and the response to treatment. To adjust the prolactin profile of a mammal, the administration of any of the substances or both substances that alter prolactin can be continued for a sufficient time to re-establish the circadian rhythm of prolactin in plasma to the phase and amplitude to that of a healthy subject of the same sex and species to which the treatment can be discontinued with the time. If the subject suffers recurrence, treatment can be resumed to adjust the prolactin profile of the subject to adjust or approximate the prolactin profile of a healthy subject of the same sex and species. The time required for the restoration varies, but it is generally within the scale of one year to one year. For some patients (for example, patients in particularly deteriorated physical condition, or those of an advanced age), it may not be possible to re-establish their prolactin rhythm within the previous periods, so these patients may require longer treatment and even continuous, with prolactin enhancers and / or reducers. The dose and timing information discussed above are designated for bromocriptine, rnelatonin and 5-hydroxytryptophan, and will have to be altered for other agents using the dose regulation and time regulation methodology described in the present invention. In the practice of this invention, a prolactin reducing compound, and / or an increase in prolactin, are administered daily to a subject preferably orally, or by subcutaneous, intravenous or intramuscular injection. The reducer or enhancer can also be administered by inhalation. Dermal delivery systems, for example, skin patches, as well as suppositories and other well-known systems for administering pharmaceutical agents can also be used. The treatment usually lasts between about 3 months and approximately an average year in humans. In this way, the administration of the prolactin reducer and / or the prolactin enhancer will thus reestablish the phase and amplitude of the neural oscillators that control the body's ability to inhibit neoplastic growth to facilitate the inhibition of neoplastic growth on a long-term (for 2R example, several months or years). An improvement in the ability to inhibit neoplastic growth can be assessed by observing partial or total ablation of the neoplasm or resumption of metastatic growth after removal of a primary neoplasm. Instead of directly measuring the neoplastic load, well-known tests of tumor burden can be used (for example, tests of neoplasm-specific antigens, nuclear magnetic resonance imaging, CT scanning, X-rays, ultrasound, counting of neoplastic cells present in blood in blood samples, etc.) to evaluate the effect of the treatment with the programmed administration of the prolactin modulators. The following more specific guidelines will be followed in general to determine initially the programming of the administration of the prolactin modulator, during a treatment period of approximately 26 weeks for human subjects: (i) Administer prolactin reducers from 600 hours to 1000 hours in a dose scale sufficient to decrease daytime prolactin levels to within 1 MEE of the normal scale of prolactin diurnal levels present in humans without neoplastic disease. (ii) Administer prolactin increments before or at bedtime on a sufficient dose scale to increase serum prolactin levels to at least the normal healthy human? level without neoplastic disease. The aspect of the invention directed to the inhibition of neoplastic growth reestablishing the prolactin profile of a mammalian subject (animal or human) having an aberrant profile of prolactin to adjust or approximate prolactin profiles for healthy young members of the same species and sex (for example, the lines b exits from Figure 1), involves the administration of a prolactin reducer, or a prolactin enhancer, or both, at predetermined doses and times determined by the aberrant prolactin profile (pretreatment ) of the subject to be treated. The amounts of prolactin reducers and / or enhancers that are required to produce this modification are within the same scales discussed above, but the time (s) of administration of this prolactin modulator (s) is ( are) determined (s) in relation to how much and when the aberrant profile differs from the normal prolactin profile (baseline curve). Methods for determining the amounts and schedule of administration are also set forth in the co-pending U.S. patent application. Series No. 07 / 995,292 and its C-I-P, Ser. No. 08 / 264,558 filed June 23, 1994, both incorporated in the present invention as reference. Another method is to administer up to 4.8 mg / day of bromocriptine as follows; 0.8 mg / day during each of the first 7 days; starting on day 8 and during the following 7 days, 1.6 rng / day is administered to the patient; starting on day 15 and during the following 7 days, 2.4 mg / day is administered; starting on day 22 and during the following 7 days, 3.2 mg / day is administered; starting on day 29 and during the following 7 days, 4.0 rng / day is administered, and starting on day 36 and during the following 7 days, 4.8 rng per day is administered for 7 consecutive days. A preferred dosage form of accelerated release bromocriptine has been described in the co-pending U.S. patent application. Series No. 08 / 171,897, also incorporated as reference. The present invention is further described and will be better understood in relation to the useful examples set forth below. These non-limiting examples should be considered as illustrative only of the principles of the invention. In addition, since many modifications and changes will be made by those skilled in the art, it is not desired to limit the invention to the exact construction and operation shown and described. Accordingly, all suitable modifications and equivalents can be used and will be contemplated within the scope of the invention and the appended claims.

EXAMPLE 1 PRQLACTINE IN PLASMA IN NORMAL AND TUMOR CARRIERS (FIBROSARCOMA DE CÉLULAS EMT-6) Adult Balb / C mice from 6 to 7 weeks (average weight of 20 grams), maintained in 12-hour daily photoperiod and allowed to be fed at will, were injected subcutaneously in the back with fibrosarcoma cells (EMT-6) at a dose of 1.7 x 106 cells. A control group remained uninjected. 14 to 21 days later, when the diameter of the tumor was 6 to 9 millimeters, the animals of the control groups injected and not injected were sacrificed at 0, 4, 8, 12, 6, or 20 hours after the beginning of the period of illumination (HALO) (n = 6-8 per time point per group), and the plasma was collected to perform prolactin analysis in plasma. The prolactin concentration in plasma (FIG. 2) was measured by radioimmunoassay (RIA) using an RIA homologous kit for prolactin in mice from Dr. A. F. Parlow, Torrance, CA. The results of this experiment show that the mammals carrying the neoplasm have a prolactin profile that is disturbed with respect to that of a healthy non-neoplasm carrier mammal of the same species and sex.

EXAMPLE 2 EFFECT OF PROLACTIN PROGRAMMED INJECTION ON TUMQRAL GROWTH (FIBROSARCQMA OF CELLS EMT-6) IN MICE Balb / C Adult Balb / C mice from 6 to 7 weeks (average weight of 20 grams) were injected with 1.7 x 10 6 EMT-6 cells (rco a) in the back while nursing (from birth) in a daily photoperiod of 12 hours . The day after inoculation, the animals were divided into two groups (n = 10 per group) and injected daily with sheep prolactin (20 pg / mouse) or vehicle (control group) at 10 HALO for 10 days (Exp. 1) or 14 days (Exp. 2), and the turnoral growth was monitored by measuring the size of the tumor with calipers. The results are shown in Table 1 below: TABLE 1 * P < 0.05 against control The results of this experiment show that the administration of prolactin 10 HALO to tumor bearing mice, results in a decrease in tarnaiño of the resulting tumors. The maximum plasma prolactin value in non-tumor-bearing healthy Balb / C mice occurs at 8-12 HALO. Thus, the administration of prolactin during the period of the maximum plasma prolactin level in healthy non-tumor-bearing mice results in tumor growth. diminished. EXAMPLE 3 EFFECT OF PROGRAMMED PRQLACTINE INJECTIONS ON TUMOR GROWTH IN MICE Balb / C Balb / C mice adult males of 6 to 7 weeks were transferred from daily photoperiods of 12 hours to constant illumination for 10 days to interrupt the circadian rhythms, time in which the tumor cells EMT-6 (1.7 x 106) were injected in the butt After inoculation of the turnoral cells, the mice were divided into 7 groups (10 mice / group), and were injected daily for 10 days with sheep prolactin (20 rncg / mouse) at 0, 4, 8, 12, 16 or 20 hours after the corticosterone injection. A control group remained untreated. At the end of the treatment, the animals were subjected to a daily photoperiod of 14 hours with the period of darkness starting 2 hours after the time when the animals were used to receive the corticosterone injection during the period of 3.1 treatment. Two weeks after the end of treatment, the volume of the tumor was determined by calibrator measurements. The results are shown in Figure 4. It was found that the inhibition of turnoral growth by treatment with prolactin depends on the time of administration. It was determined that the greatest turnoral growth inhibition was in the 8-hour prolactin / corticosterone group (ie, prolactin injected 8 hours after the corticosterone injection) (85 ± 15 mrn3 for mice treated for 8 hours with prolactin / corticosterone against 350 ± 35 mrn3 for untreated mice; P.0.01). This example demonstrates that the reduction of the tumor depends to a large extent on the time of administration of prolactin with respect to the maximum level of corticosterone induced which, in the absence of a photoperiod, establishes the cire dianos rhythms of the mice.

EXAMPLE 4 EFFECT OF PROGRAMMED PROLINTIN INJECTION ON METASTATIC DISEMINATION IN TUMOR CARRIER MICE Male C57 black male mice from 6 to 7 weeks were transferred from 12-hour photoperiods to constant illumination for 7 days when they were injected into the leg bearing with LL-2 cells from Le? Is lung carcinoma (1 x 10 ^ /mouse). The mice were kept in a constant photopepod for the duration of the treatment »Approximately 3 weeks after the injection of the tumor cells when the primary tumor was 7 m diameter, it was surgically removed and the mice were divided into 7 groups (5 to 7 mice / group), and were injected with prolactm (20 rncg / mouse) at 4, 8, 12, 16 or 20 hor-s after the cortical injection. erona (20 rncg / mouse) for 10 days. A control group remained untreated. After the treatment ended, the animals were subjected to a daily photopepod of 12 hours, with the period of darkness starting 2 hours after the animals were used to receive corticosteroid injection during the treatment period. Three days after the end of the treatment, the mice were sacrificed to determine rnetastatic spread towards the lung (determined by the weight of the lung). The results are shown in Table 2.

TABLE 2 PULMON WEIGHT OF MICE CONTROL OR MICE INJECTED WITH CELLS OF THE LUNG PULMONARY CARCINOMA 0 0 In healthy C57 black mice subjected to a constant photoperiod, the secretion of prolactin reaches a maximum at 0 hours after the maximum level of - > K corticosterone. The results of this experiment show that maximal inhibition of tumor growth is achieved by injecting prolactin within 0 to 4 hours after the maximum level of corticosterone, that is, at the same time as prolactin reaches a maximum after injection of prolactin. corticosterone in healthy C5 black mice subjected to a constant photoperiod (normal profile of prolactin in black C5 mice). Thus, programmed prolactin injections that occur at the same point in a circadian cycle in which prolactin levels reach maximally in healthy animals of the same species and sex, can significantly decrease the degree of metastatic growth after What is the primary tumor removed? EXAMPLE 5 EFFECT OF SCHEDULED BROMQCRIPTINE ADMINISTRATION ON TUMOR GROWTH (FIBROSRRCOMA OF EMT-6 CELLS) IN MICE Balb / C Balb / C mice adult males of 6 to 7 weeks subjected to a daily photoperiod of 12 hours, are injected with tumor cells EMT-6 (1.7 x 106) in the rear. After inoculation of the turnoral.es cells, the mice are divided into 7 groups (10 mice / group). Three groups are injected daily for 10 days with bromocriptine (50 rncg / mouse at 0, 12 and 20 hours after the start of the lighting period.) Three groups (control) receive only one injection of vehicle at the same time (0, 12 and 20 HALO) .A control group remains untreated.Two weeks after the end of treatment, tumor volume is determined by calibrator measurements.Tumor growth will be inhibited by the administration of brornocriptine.Maximum inhibition of tumor growth by treatment with bromocriptine will occur in those mice injected with bromocriptine at 0 hours after the start of the illumination period.The maximum level of prolactin occurs at 8-12 HALO.This corresponds to the prolactin profile of healthy Balb / C mice, as shown in Figure 2.

EXAMPLE 6 EFFECT OF THE PROGRAMMED ADMINISTRATION OF BROMOCRIPTIN AND MELATONIN ON THE GROWTH TUMQRAL FIBROSARCOMA OF CELLS EMT-6) IN MICE Balb / C B ib / C mice, adult males 6 to 7 weeks old, are injected with ternary EMT-6 cells (1.7 x 106) in the posterior. After the inoculation of the cells. urnorales, the mice are divided into 8 groups (10 mice / group), and injected daily for 10 days with broinocppt na at 0 HALO, the time determined in Example 5 which results in greater inhibition of tumor growth (50 rncg / mouse). Mice are also injected with melatonin (40 ng / g / mouse) at 0, 4, 8, 12, 16, or 20 hours after the injection of brornocriptine. A control group remains untreated, and another control group is treated only with b omocrip-. na Two weeks after the end of treatment, the volume of the tumor is determined by gauge-measurements. It is found that tumor growth is inhibited to a greater extent by the combination of the scheduled administration of brnocryptine at 0 HALO and rnelatomine at 12 hours after the injection of brornocnptin, than by the scheduled administration of bromocriptine alone, and q? E the improvement of the turnoral growth inhibition by treatment with nelatonin depends on the time of the administration of the same. The maximum effect of melatoni a is at 12 HALO because this stimulates the release of prolactin at the time of day when prolactin exhibits the most inhibitory activity against neoplastic growth, and it is also the time at which levels of melatonin reach a maximum in healthy mice not carriers of metastasis. The method of the present invention can be used to treat a broad spectrum of neoplastic diseases which include, but are not limited to, sarcoma, rosarcorna, glioblastoma, carcinoma, rnelanoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, leukemia, and other neoplastic conditions.

Claims (21)

NOVELTY OF THE INVENTION CLAIMS

1. The use of a prolactin reducer for the preparation of a medicament for inhibiting neoplastic growth in a mammal, said medicament being administered to said mammal at a predetermined time during a period of 24 hours.

2. The use according to claim 1, further characterized in that said prolactin reducer is a dopamine agonist.

3. The use according to claim 2, further characterized in that the predetermined time is between around 05:00 hr and approximately 13:00 hr.

4. The use according to claim 1, further characterized in that the medicament is administered to adjust the prolactin level of said mammal at a predetermined time when the prolactin level at the time of day of said mammal is greater than 1 SEM. above the baseline prolactin profile at two separate time points or when the prolactin level of the daytime of said mammal is greater than 2 SEM above the prolactin profile of baseline at a point in time.

5. The use according to claim 3, further characterized in that said prolactin reducer is bromocriptine.

6. The use according to claim 5, further characterized in that said mammal is a human and the drug is administered in such amount. that bromocripti assorted falls within the range of 0.8-8.0 rng / day.

7. The use according to claim 6, further characterized in that the predetermined time is between about 05:00 hr and approximately 13:00 hr.

8. The use according to claim 1, further characterized in that said neoplasm is a member selected from the group consisting of sarcomas, fibrosarcoma, carcinomas, glblastomas and melanoma.

9. The use of a prolactin reducer for the preparation of a medicament for inhibiting the growth of neoplasms in a mammal having a prolactin profile, wherein said medicament is administered to adjust the prolactin profile of said mammal carrying the prolactin. neoplasm to conform or approach the normal prolactin profile, thus inhibiting the growth of said neoplasm.

10. The use according to claim 9, further characterized in that said prolactin reducer is a dopamine agonist.

11. The use according to claim 10, further characterized in that the predetermined time is between about 05:00 hr and about 13:00 hr.

12. The use according to claim 11, further characterized in that said prolactin reducer is brornocriptine, said mammal is a human being, and the medicament is administered in an amount such that bromocriptine is administered within the range of 0.8- 8.0 rng / day.

13. The use according to claim 1, further characterized in that the prolactin reducer is used together with a prolactin enhancer for the preparation of a medicament for the treatment of said mammal.

14. The use according to claim 13, further characterized in that said medicament containing prolactin enhancer is administered at a predetermined time during a period of 24 hours.

15. The use according to claim 14, further characterized in that said prolactin enhancer is rnelatonin and the predetermined time is at bedtime.

16. The use according to claim 15, further characterized in that said mammal is a human and said drug containing rnelatonin is administered in such a manner that the rnelatonin is administered in an amount within the range of 1.0 to 20.0 rng / 'day.

17. The use according to claim 16, further characterized in that said prolactin reducer is rornocriptine.

18. - The use according to claim 17, further characterized because the romocriptma is administered between about 05:00 hr and about 10:30 hr and is administered in such a way that the broinocpptina is adirnin strada in an amount within the range of 0.8-8.0 g / day.

19. The use according to claim 9, characterized in that the prolactum reducer is used together with a prolactin enhancer for the preparation of a medicament for the treatment of said mammal.

20. The use according to claim 13, further characterized in that said prolactum enhancer is rnelatonma and said prolactum reducer is brornocriptine.

21. The use according to claim 19, further characterized in that said prolactin enhancer is rnelatonin and said prolactum reducer is brornoeript i na.