PHARMACEUTICAL COMPOSITIONS CONTAINING 1-ISOPROPYLDIHYDROLYSERGIC ACID

Abstract: This invention relates to the use in vivo of 1-isopropyldihydrolysergic acid as a 5-HT₂ antagonist.

Description: Garbrecht et al ., U.S. Patent No. 4,714,704, and Cohen et al ., U.S. Patent No. 4,713,384, describe dihydrolysergic acid esters of the following formula The group R is primary or secondary C₁₋₈ alkyl, CH₂-C₂₋₄ alkenyl, C₃₋₈ cycloalkyl, or C₃₋₆ cycloalkyl substituted C₁₋₅ primary or secondary alkyl, the total number of carbon atoms not exceeding 8, and the group R¹ is C₁-C₄ straight chain alkyl. In Garbrecht et al ., the group R² is C₁₋₃ alkyloxy C₅₋₇ cycloalkyl, whereas, in Cohen et al ., it is hydroxy C₅₋₇ cycloalkyl. Highly preferred compounds are those in which R is isopropyl and R¹ is methyl. These compounds, all esters, are recognized to possess selective and highly potent antagonist activity at 5HT₂ (5HT = serotonin) receptors upon intravenous or oral administration in rats. Both of the foregoing classes of esters correspondingly demonstrate, as expected, high affinity in vitro for 5HT₂ receptors. These properties are to be compared to the in vitro affinity of a corresponding free acid, 1-isopropyldihydrolysergic acid, i.e., in the foregoing formula, R is isopropyl, R¹ is methyl, and R² is hydrogen. The latter compound is recognized to exhibit a relatively low level of affinity in vitro for 5HT₂ receptors. It has been assumed, therefore, that the free acid would not be a likely candidate for use as an antagonist at 5HT₂ receptors, and the literature suggests the utility of the free acid compounds solely as intermediates to active compounds. In this regard, see, e.g., the earlier cited Garbrecht et al . and Cohen et al ., as well as Garbrecht, U.S. Patent No. 3,580,916; Rucman, U.S. Patent No. 4,230,859; and Hofmann et al ., U.S. Patent No. 3,249,617. Surprisingly, however, we have recently discovered that 1-isopropyldihydrolysergic acid exhibits relatively high potency as a 5-HT₂ antagonist in vivo irrespective of its relatively low affinity for these receptors when measured in vitro . It is thus to the use of 1-isopropyldihydrolysergic acid as a 5HT₂ antagonist, whether administered orally or by injection, that this invention is directed. Thus, this invention is directed to a method of blocking 5HT₂ receptors which comprises administering to a mammal having an excess of serotonin, whether centrally or peripherally, a 5HT₂ blocking dose of a compound of the formula or any of its pharmaceutically acceptable salts. It is recognized that a method whereby 5HT₂ receptors are blocked but alpha receptors are not affected by a particular compound at a given dose level is potentially useful in treating disease states in which an excess of circulating serotonin is a major cause. These disease states include hypertension, anorexia nervosa, depression, mania, carcinoid syndrome, migraine, sexual dysfunction, thrombosis, ischemia, and vasospasm. The lack of alpha receptor inhibitory activity indicates that the usual undesirable side effects associated with alpha receptor blockade--postural hypotension, tachycardia, impotence, and increase plasma renin levels-- will not occur. This lack of undesirable side effects is in marked contrast to many presently available hypotensive agents, including ketanserin. The compound may be administered in the form of its pharmaceutically acceptable salts. Pharmaceutically-acceptable acid addition salts include salts derived from non-toxic inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydriodic acid and phosphorous acid, as well as salts derived from non-toxic organic acids such as aliphatic mono- and dicarboxylic acids, phenyl-substituted alkanoic acids, hydroxy alkanoic acids, alkanedioic acids, aromatic acids, aliphatic and aromatic sulfonic acids. Examples of such pharmaceutically-acceptable salts thus are sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate, chloride, bromide, iodide, fluoride, acetate, propionate, decanoate, caprylate, acrylate, formate, isobutyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, mandelate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methoxybenzoate, phthalate, terephthalate, benzenesulfonate, toluenesulfonate, chlorobenzenesulfonate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, β-hydroxybutyrate, glycollate, malate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate and naphthalene-2-sulfonate. In addition, salts of pharmaceutically acceptable alkali and alkaline earth metals may be used. These include, for example, sodium, potassium, lithium, calcium and magnesium. In carrying out the therapeutic method of this invention, a pharmaceutically acceptable salt of 1-isopropyldihydrolysergic acid may be formed, and the resulting salt then administered orally or parenterally to a mammal, preferably human, having an excess of circulatory serotonin and for whom it is desirable to block 5HT₂ receptors to alleviate symptoms such as high blood pressure and migraine, attributable to excessive serotonin levels. For parenteral administration, a water soluble salt of the compound may be dissolved in an isotonic salt solution and administered, for example, intravenously. For oral administration, a pharmaceutically acceptable salt of the compound may be mixed with standard pharmaceutical excipients such as starch and the mixture loaded into capsules or made into tablets, each containing preferably 0.001 to 100 mg of active compound. Dosage levels of from 0.001 to 10 mg/kg are effective in blocking 5HT₂ receptors. Preferably, an oral dose is administered 2 to 4 times per day, providing a daily dosage in the range of 0.003 to 10.0 mg/kg per day. A variety of oral dosage forms, including suspensions, elixirs and tablets can also be utilized and are preparable by standard procedures. The following examples are provided to illustrate the present invention. Example 1 -- In Vitro 5HT₂ Affinity of 1-Isopropyldihydrolysergic Acid. Male Wistar rats (150-300 g) (Harlan Sprague-Dawley, Inc., Cumberland, IN) were killed by cervical dislocation. Ring preparations of external rat jugular veins were used as detailed in [(Cohen et al ., J. Pharm. and Exper. Therapeutics 227 , 327-332 (1983); Cohen et al ., J. Pharm. and Exper. Therapeutics 235 , 319-323 (1985).] Tissues were mounted in organ baths containing 10 ml of modified Krebs' solution of the following composition (millimolar concentrations): NaCl, 118.2; KCl, 4.6; CaCl₂·2H₂O, 1.6; KH₂PO₄, 1.2; MgSO₄, 1.2; dextrose, 10.0; and NaHCO₃, 24.8. Tissue bath solutions were maintained at 37°C and aerated with 95% O₂-5% CO₂. Isometric contractions were recorded as changes in grams of force on a Beckman Dynograph with Statham UC-3 transducers and microscale accessory attachments. Tissues were allowed to equilibrate at an initial optimum resting force for 1 to 2 hr before exposure to test compound. After control contractile responses to serotonin were obtained in the jugular vein, vessels were incubated with appropriate concentrations of the test compound for 1 hr. Responses to serotonin were then repeated in the presence of test compound. Apparent test compound dissociation constants (K B ) were determined for each concentration of test compound according to the following equation: K B = (B) (dose ration - 1) where (B) is the concentration of the test compound and dose ratio is the ED₅₀ of the serotonin in the presence of the test compound divided by the control ED₅₀. These results were then expressed as the negative logarithm of the K B ( i.e. , -log K B ). The data were also analyzed according to the procedure of Arunlakshana and Schild, Brit. J. Pharmacol. 14 , 48-58 (1959) as previously described [Cohen et al ., supra (1983)]. Table I following depicts apparent dissociation constants derived from the ability of 1-isopropyldihydrolysergic acid and that of the corresponding 4-methoxycyclohexyl ester to antagonize serotonin-induced contractions in the rat jugular vein. Affinity at 5HT₂ receptors for 1-isopropyldihydrolysergic acid was approximately 10⁻⁷M. Furthermore, this agent was a competitive antagonist at 5HT₂ receptors in the jugular vein as indicated by a slope of the Schild plot that did not differ from negative unity. The free acid was a competitive antagonist at 5HT₂ receptors, albeit at an affinity approximately 100-fold lower than that of the 4-methoxycyclohexyl ester. Example 2 -- In Vivo Vascular 5HT₂ Blocking Activity of 1-Isopropyldihydrolysergic Acid. Serotonin receptor antagonism was evaluated in pithed Wistar normotensive rats (Charles River, Inc. Portage, MI; 240-374 g) since responses in the pithed preparation are primarily direct vascular effects. Rats were anesthetized with Metofane® (methoxyflurane), carotid arterial and femoral venous catheters were implanted, and the trachea was cannulated. Rats were pithed and ventilated with room air (Cohen et al ., supra , 1983; Cohen et al ., supra , 1985). An equilibration period of 15 min. was observed before control measurements and i.v. administration of test compound or vehicle. Increasing intravenous doses of serotonin were injected 15 min. after intravenous administration of test compound or vehicle. As shown in Table II following, administered intravenously, both 1-isopropyldihydrolysergic acid and the corresponding 4-methoxycyclohexyl ester dose-dependently inhibited the pressor response to serotonin in pithed rats. By intravenous administration, 1-isopropyldihydrolysergic acid was only approximately one-third as potent as the ester in blocking vascular 5HT₂ receptors. Since 1-isopropyldihydrolysergic acid required only a three-fold higher intravenous dose than the corresponding 4-methoxycyclohexyl ester to block vascular 5HT₂ receptors, a comparison of these compounds with regard to duration of effective blockade of vascular 5HT₂ receptors in vivo was conducted. For these studies pithed rats were treated with vehicle or with the acid or the ester at a dose of 0.1 mg/kg, i.v. and challenged with serotonin (0.1 mg/kg i.v.) at various intervals for 2 hrs. after test compound or vehicle administration. As seen previously, the ester (0.1 mg/kg, i.v.) produced greater inhibition of vascular serotonin receptors than the acid (0.1 mg/kg, i.v.); nevertheless, the acid did produce a marked block of vascular serotonin receptors. Response to serotonin after both the ester and acid administration slowly returned toward saline values although marked inhibition of serotonin vascular response was still apparent 2 hrs. after intravenous administration of either agent. The results are shown in Table III following.

Claims: Claims for the following Contracting Sates : AT, BE, CH, DE, DK, FR, GB, IT, LI, NL, SE 1. A pharmaceutical formulation comprising, as active ingredient, a compound of the formula or any of its pharmaceutically acceptable salts, associated with one or more pharmaceutically-acceptable carriers therefor. 2. A compound of the formula or a pharmaceutically acceptable salt thereof, for use in blocking 5HT₂ receptors. 3. A compound of the formula or a pharmaceutically acceptable salt thereof, for use in treating hypertension. 4. A compound of the formula or a pharmaceutically acceptable salt thereof, for use in treating migraine. 5. A compound of the formula or a pharmaceutically acceptable salt thereof, for use in treating vasospasm. 6. A compound of the formula or a pharmaceutically acceptable salt thereof, for use in treating depression. Claims for the following Contracting States: GR, ES 1. A pharmaceutical formulation comprising, as active ingredient, a compound of the formula or any of its pharmaceutically acceptable salts, associated with one or more pharmaceutically-acceptable carriers therefor. 2. A process for preparing a formulation, which comprises admixing, as active ingredient, a compound of the formula or any of its pharmaceutically acceptable salts, with one or more pharmaceutically-acceptable carriers therefor.

Inventors: COHEN, MARLENE LOIS, GOLDBERG, MARK JAY, LEMBERGER, LOUIS
Applicants: ELI LILLY AND COMPANY
Application year: 1990
Main classification: A61K31
IPC codes: A61P43, A61P25, A61P9, C07D457, A61K31