KR100491062B1 - Calcium receptor active arylalkylamine - Google Patents

Abstract

The present invention relates to molecules having the following general formulas (I) and (II) which induce or block inorganic ion receptor activity. Preferably, this molecule can mimic or block the effect of extracellular Ca ²⁺ on calcium receptors. A preferred use of this molecule is to treat a disease or condition by altering inorganic ion receptor activity, preferably calcium receptor activity.

Description

Calcium Receptor Active Arylalkyl Amine

Field of invention

The present invention relates to the design, development, composition and use of new molecules capable of modulating the activity of inorganic ion receptors.

Background of the Invention

Certain cells in the body respond not only to chemical signals but also to ions such as extracellular calcium ions (Ca ²⁺ ). Changing the concentration of extracellular calcium Ca ²⁺ (herein referred to as “[Ca ²⁺ ]”) alters the functional response of these cells. One such differentiated cell is an epithelial cell that secretes epithelial hormone (PTH). PTH is a major endocrine factor that controls Ca ²⁺ homeostasis in blood and extracellular fluids.

PTH acts on bone and kidney cells to increase the level of Ca ²⁺ in the blood. This increase in [Ca ²⁺ ] then acts as a negative feedback signal, inhibiting PTH secretion. The interrelationship between [Ca ²⁺ ] and PTH secretion forms an essential mechanism for maintaining Ca ²⁺ homeostasis in the body.

Extracellular Ca ²⁺ acts directly on epithelial cells to regulate PTH secretion. The presence of an epithelial cell surface protein that detects a change in [Ca ²⁺ ] has been identified (see Brown et al. 366 Nature 574, 1993). In epithelial cells, this protein acts as a receptor for Ca ²⁺ in the cell (“calcium receptor”) and senses a change in [Ca ²⁺ ] to initiate PTH secretion, a functional cellular response.

Extracellular Ca ²⁺ may affect other cell functions, as reviewed in Nemeth et al. (11 Cell Calcium 319, 1990). The role of extracellular Ca ²⁺ in paracytic cells (C-cells) and epithelial cells is discussed in Nemed et al. (11 Cell Calcium 323, 1990). These cells have been shown to express similar Ca ²⁺ receptors [Brown et al. 366 Nature 574, 1993; 9 Suppl by Mital et al. 1 J. Bone and Mineral Res. s282, 1994; 9 Suppl by Rogers et al. 1. J. Bone and Mineral Res. s409, 1994; 9 Suppl, Garrett et al. 1. J. Bone and Mineral Res. s409, 1994]. The role of extracellular Ca ²⁺ on osteoclasts is discussed in Zaidi's [10 Bioscience Reports 493, 1993]. Keratinocytes, glomeruli cells, tropoblasts, pancreatic beta cells and adipocytes all respond to an increase in extracellular calcium which likewise affects the activation of calcium receptors of these cells.

The ability of various compounds to mimic extracellular Ca ²⁺ in vitro is described in Nemeth et al., “Calcium-Binding Proteins in Health and Disease” (spermine and spermidine), 1987, Academic Press, Inc. , pp. 33-35; Brown et al. (Eg neomycin) 128, Endocrinology 3047, 1991; Chen et al., (Diltiazem and its analogs, TA-3090). 5 J. Bone and Mineral Res. 581, 1990; And (dyrapamine) 167 Biochem. Biophys. Res. 807, 1990. PCT / US93 / 01642 of Nemed et al., International Publication No. WO 94/18959 and PCT / US92 / 07175 of Nemed et al., WO 93/04373, disclose for cells with inorganic ion receptors. Disclosed are various compounds that can modulate the effects of inorganic ions, preferably the effects of calcium on calcium receptors.

The references provided in the background of the invention are not recognized as prior art.

Summary of the Invention

It is a feature of the present invention to molecules that can modulate the activity of one or more inorganic ion receptors. Preferably, the molecule can mimic or block the effects of extracellular Ca ²⁺ on calcium receptors. Preferred use of such molecules is to treat a disease or condition by altering inorganic ion receptor activity, preferably calcium receptor activity.

Extracellular Ca ²⁺ is under tight homeostatic control and regulates a variety of processes, such as retention, nerve and muscle excitability, and proper bone formation. Calcium receptor proteins allow certain differentiated cells to respond to changes in extracellular Ca ²⁺ concentrations. For example, extracellular Ca ²⁺ inhibits the secretion of epithelial hormones from epithelial cells, inhibits bone uptake by osteoclasts, and stimulates the release of calcitonin from C-cells.

Compounds that modulate inorganic ion receptor activity can be used to treat a disease or condition by affecting one or more of the activities of the inorganic ion receptor, thus providing a beneficial effect to the patient. For example, osteoporosis is an age-related disease characterized by a loss of bone mass and an increased risk of fracture. Directly (eg osteoclast ion mimic compounds) or indirectly (eg C-cell ion mimics) by increasing endogenous calcitonin levels, and / or by reducing epithelial hormone levels (eg, Epithelial cell ion mimics) Osteoclasts Compounds that block bone resorption delay bone loss and thus provide a beneficial effect in patients with osteoporosis.

Intermittently low doses of PTH are known to cause anabolic effects on bone mass and proper bone remodeling. Thus, compounds that cause a transient increase in epithelial hormones and dosing control (eg, intermittent administration of epithelial cell ionic degradation) can increase bone mass in patients with osteoporosis.

In addition, diseases or disorders characterized by a deficiency of one or more inorganic ion receptor activities can be treated by the present invention. For example, certain forms of primary hyperthyroidism are characterized by abnormally high levels of epithelial hormones and reduced epithelial gland sensitivity to circulating calcium. Calcium receptor modulators can be used to modulate the sensitivity of epithelial cells to calcium.

Preferably, the compound is used to treat a disease or condition that may be affected by modulating calcium receptor activity and modulating one or more activities of the calcium receptor. Preferably, the disease or condition is directed to abnormal bone and mineral homeostasis, more preferably calcium homeostasis.

Abnormal calcium homeostasis is characterized by one or more of the following activities: (1) abnormal increase or decrease in serum calcium, (2) abnormal increase or decrease in urine secretion of calcium, (3) eg bone Abnormal increase or decrease in bone calcium levels as assessed by mineral density measurements, (4) abnormal absorption of food calcium and (5) production of circulating carriers or hormones that affect calcium homeostasis such as epithelial hormone and calcitonin, and (Or) abnormal increase or decrease in release. Abnormal increases or decreases in these different aspects of calcium homeostasis are those that occur in the general population and are generally associated with a disease or condition.

More generally, molecules that modulate the activity of inorganic ion receptors are useful in the treatment of diseases characterized by abnormal inorganic ion homeostasis. Preferably, such molecules modulate one or more actions of the inorganic ion receptor. Inorganic ion receptor modulators include ion mimics, ion degrading, calcium mimicking and calcium degrading.

Ion mimics are molecules that mimic the effect of increasing ion concentration in inorganic ion receptors. Preferably, this molecule affects one or more calcium receptor activities. Calcium mimetics are ion mimics that affect one or more calcium receptor activities and preferably bind calcium receptors.

Ionolytic forms are molecules that reduce or block one or more activities caused by inorganic ions on an inorganic ion receptor. Preferably, this molecule inhibits one or more calcium receptor activities. The calcium degrading type is an ionolytic type that inhibits one or more calcium receptor activities induced by extracellular calcium and preferably binds to the calcium receptor.

Inorganic ion receptor modulators may be formulated into pharmacological agents or compositions to facilitate administration to a patient. A pharmacological agent or composition is an agent or composition suitable for administration to a mammal, preferably a human. Considerations for suitable forms of administration are known in the art and include toxic effects, solubility, mode of administration and maintenance activity.

Thus, in a first aspect, a feature of the present invention is an inorganic ion receptor modulator consisting of molecules that induce or block one or more inorganic ion receptor activities produced by extracellular inorganic ions. The molecule has the following general formula:

here,

X is independently selected from the group consisting of isopropyl, CH ₃ O, CH ₃ S, CF ₃ O, an aliphatic ring and an attached or fused aromatic ring,

m is 0-5 each independently.

Preferably, the aromatic and aliphatic rings have 5 to 7 elements. More preferably, aromatic and aliphatic rings have only carbon atoms (ie this ring is not a heterocyclic ring).

Preferably, the molecule induces or blocks one or more calcium receptor activities caused by extracellular calcium.

Another feature of the present invention lies in an inorganic ion receptor modulator having the general formula:

here,

X is independently H, CH ₃ , CH ₃ O, CH ₃ CH ₂ O, methylenedioxy, Br, Cl, F, CF ₃ , CHF ₂ , CH ₂ F, CF ₃ O, CH ₃ S, OH, CH ₂ OH, CONH ₂ , CN, NO ₂ , CH ₃ CH ₂ , propyl, isopropyl, butyl, isobutyl, t-butyl, acetoxy, an aliphatic ring and an attached or fused aromatic ring,

Each R is independently hydrogen, methyl, ethyl, propyl, isopropyl, butyl, allyl, isobutyl, t-butyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, indenyl, indanyl, dihydroindolyl, Thiodihydroindolyl, and 2-, 3- or 4-piperidyl (dinyl), and

m is 0-5 each independently.

The molecule induces or blocks one or more inorganic ion receptor activities produced by extracellular inorganic ions. Preferably, the molecule induces or blocks one or more calcium receptor activities caused by extracellular calcium.

In a preferred embodiment, R is H, CH ₃ , ethyl or isopropyl, each X is independently a group consisting of isopropyl, CH ₃ O, CH ₃ S, CF ₃ O, an aliphatic ring and an attached or fused aromatic ring Is selected from. Preferably, the aliphatic ring and the attached or fused aromatic ring have 5 to 7 elements. More preferably, the aromatic ring and aliphatic ring contain only carbon atoms.

Another feature of the present invention is an inorganic ion receptor comprising a molecule selected from the group consisting of Compound 4L, Compound 8J, Compound 8U, Compound 9R, Compound 11X, Compound 12U, Compound 12V, Compound 12Z, Compound 14U, Compound 16M and Compound 16P. It is in the regulator.

Another feature of the present invention is a method of using the formulations disclosed herein to treat a disease or condition by modulating inorganic ion receptor activity. Patients in need of such treatment may use inorganic medical ions that affect inorganic ion homeostasis by detecting a deficiency of proteins whose production and secretion are affected by standard medical techniques, such as general blood analysis, for example, by changing inorganic ion concentrations. Or by detecting abnormal levels of hormones.

Therapy involves the administration of a therapeutically effective amount of an inorganic ion receptor modulator to a patient. In a preferred embodiment, the method is used to treat a disease or condition characterized by abnormal inorganic ion homeostasis, more preferably a disease or condition characterized by abnormal calcium homeostasis. Diseases or disorders characterized by abnormal calcium homeostasis include epithelial hyperplasia, osteoporosis, and other bone and mineral related disorders (see, for example, standard medical references such as "Harrison's Principles of Internal Medicine"). Included. These diseases and disorders are treated with calcium receptor modulators that mimic or block one or more effects of Ca ²⁺ , thereby directly or indirectly affecting the levels of proteins or other molecules in the body of the patient.

A "therapeutically effective amount" means the partial or complete recovery of one or more physiological or biochemical factors that relate to or cause the disease or condition to some extent or alleviate one or more symptoms of the patient's disease or condition. It means the amount of the formulation to make.

In a preferred embodiment, the patient has a disease or condition characterized by abnormal levels of one or more of the components regulated by calcium receptors, the molecule comprising epithelial cells, osteoclasts, glomerular kidney cells, myocardial renal cells , Distal tubular kidney cells, central nervous system cells, peripheral nervous system cells, thick ascending cells of the Henle's loop and / or collecting ducts, keratinocytes of the epidermis, paracytic cells of the thyroid (C-cells), intestinal cells, Placental tropoblast, platelets, vascular smooth muscle cells, ventricular cells, gastrin secreting cells, glucagon secreting cells, renal vessel membrane cells, breast cells, beta cells, adipocytes, immune cells and GI vascular cells It is active against calcium receptors.

More preferably, the cell is an epithelial cell, the molecule lowers the level of epithelial hormone in the serum of the patient, even more preferably this level is lowered enough to reduce plasma Ca ²⁺ , Most preferably, epithelial hormone levels are lowered to levels present in normal individuals.

Accordingly, a feature of the present invention is in agents and methods useful for treating such diseases or disorders by modulating inorganic ion receptor activity. For example, the molecules of the invention can be used to target calcium receptors for different cell types that sense and respond to changes in external calcium. For example, molecules that mimic external calcium can be used to selectively inhibit the secretion of epithelial hormones from epithelial cells or to inhibit bone uptake by osteoclasts, or to stimulate the release of calcitonin from C-cells. . This molecule can be used to treat diseases or disorders characterized by abnormal calcium homeostasis, such as epithelial hyperplasia and osteoporosis.

Other features and advantages of the invention will be apparent from the following detailed description of preferred embodiments of the invention and from the claims.

Brief description of the drawings

1A-D, 2A-D, 3A-E, 4A-E, 5A-D, 6A-E, 7A-E, 8A-E, 9A -F, 10A-E, 11A-E, 12A-D, 13A-D, 14A-D, 15A-D, 16A-D, 17A-D FIGS. 18A-E, 19A-D and 20A-D are derived from diphenylpropyl-α-phenethylamine illustrating a group of molecules prepared and screened to find useful molecules of the invention. Chemical structure of the molecule.

Detailed Description of the Preferred Embodiments

The present invention describes inorganic ion receptor modulators that can mimic or block the effects of inorganic ions on inorganic ion receptors. A preferred use of inorganic ion receptor modulators is to treat a disease or condition by modulating inorganic ion receptor activity. Preferably these molecules are used to treat a disease or condition characterized by abnormal ion homeostasis, more preferably abnormal calcium homeostasis. Other uses of inorganic ion receptor modulators such as diagnostics are known in the art (see PCT / US93 / 01642 to Nemed et al. And WO 94/18959).

I. Calcium Receptors

Calcium receptors and nucleic acids encoding them are disclosed in PCT / US93 / 01642 to Nemed et al. And WO94 / 18959. Calcium receptors are epithelial cells, osteoclasts, glomerular kidney cells, mesothelial tubular kidney cells, distal tubular kidney cells, central nervous system cells, peripheral nervous system cells, narcotic and / or thick ganglion cells of the collecting duct, keratinocytes of the epidermis , Thyroid cells (C-cells), intestinal cells, placental tropoblasts, platelets, vascular smooth muscle cells, ventricular cells, gastrin secreting cells, glucagon secreting cells, renal vessel membrane cells, breast cells. It is present in different cell species such as beta cells, adipocytes, immune cells and GI duct cells. The calcium receptors present in these cell types may differ from one another. In addition, the cells may have one or more calcium receptors.

Comparing the calcium receptor activity and amino acid sequence of different cells, it can be seen that different calcium receptor species exist. For example, calcium receptors can respond to a variety of bi- and trivalent cations. Epithelial calcium receptors respond to calcium and Gd ³⁺ , while osteoclasts respond to divalent cations such as calcium but not to Gd ³⁺ . Thus, epithelial calcium receptors are pharmacologically different from calcium receptors on osteoclasts.

On the other hand, nucleic acid sequences encoding calcium receptors present in epithelial cells and C-cells indicate that these receptors have very similar amino acid structures. Nevertheless, calcium mimetic compounds exhibit differential pharmacology and modulate different activities in epithelial cells and C-cells. Thus, the pharmacological properties of calcium receptors may vary significantly depending on the cell species or organs on which these receptors are expressed even if they have a similar structure.

Calcium receptors generally have low affinity for extracellular Ca ²⁺ (apparent K _d is generally at least about 0.5 mM). Calcium receptors are described in Cooper, Bloom and Roth, "The Biochemical Basis of Neuropharmacology," Ch. And free or bound effector mechanisms as defined in 4], and are therefore distinguished from intracellular calcium receptors such as calmodulin and troponin.

Calcium receptors respond to changes in extracellular calcium levels. Its exact change depends on the specific receptor and the cell line containing the receptor. For example, the in vitro actions of calcium on calcium receptors in epithelial cells include:

1. An increase in internal calcium. This increase is due to the influx of external calcium and / or the mobilization of internal calcium. Features of increased internal calcium include the following:

(a) Rapid [Ca ²⁺ ] ₁ disappearance by pretreatment with ionomycin (if no extracellular Ca ²⁺ is present) without responding to inhibition of 1 μM La ³⁺ or 1 μM Gd ³⁺ . And a temporary increase (peaking time: less than 5 seconds).

(b) this increase is not inhibited by dihydropyridine.

(c) Temporary increase disappeared by pretreatment with 10 mM sodium borate for 10 minutes.

(d) Temporary increase is reduced by pretreatment with protein kinase C (PKC) activator such as phorbol myristate acetate (PMA), mazelein or (-)-indoracam V. The overall effect of protein kinase C activators is to shift the concentration-response curve for calcium to the right without affecting maximal response.

(e) Treatment with Pertussis toxin (greater than 4 hours at 100 ng / ml) did not affect this increase.

2. Rapid (less than 30 seconds) increase in the formation of inositol-1,4,5-triphosphate or diacylglycerol. Treatment with Pertussis toxin (greater than 4 hours at 100 ng / ml) does not affect this increase.

3. Inhibition of cyclic AMP formation stimulated with dopamine- and isoproterenol. This effect is blocked by pretreatment with Pertussis toxin (greater than 4 hours at 100 ng / ml).

4. Inhibition of PTH secretion. Treatment with Pertussis toxin (greater than 4 hours at 100 ng / ml) does not affect the inhibition of PTH secretion.

Techniques known in the art can be used to readily determine the effect of calcium on different calcium receptors of different cells. This effect may be similar in the increase in internal calcium observed in epithelial cells. However, this effect is expected to be different in other respects, for example, causing or inhibiting the release of hormones other than the epithelial hormone.

II. Inorganic ion receptor modulators

Inorganic ion receptor modulators cause or block one or more inorganic ion receptor activity caused by extracellular inorganic ions. Calcium receptor modulators can mimic or block the effects of extracellular Ca ^{2+ on} calcium receptors. Preferred calcium receptor modulators are calcium mimetics and calcium degradation. The intrinsic and specific structures of the inorganic ion receptor modulators are provided in the Summary and FIG.

Inorganic ion receptor modulators can be distinguished by selecting molecules that have been shown to have unique activity, ie, molecules that are modeled after the lead molecule (see, eg, PCT / US93 / 01642 to Nemed et al. And WO 94/18959). .

Preferred inorganic ion receptor modulators disclosed by the present invention are compounds 8J, 8U, 9R, 11X, 12U, 12V, 12Z, 14U, 16M and 16P. These compounds all have EC ₅₀ values of less than 5 μM.

The EC ₅₀ value is the concentration of the molecule causing half of the maximum effect. The IC ₅₀ value is the concentration of the molecule causing half of the maximum blocking effect. EC ₅₀ values or IC ₅₀ values can be determined by analyzing one or more of the activities of the inorganic ions at the inorganic ion receptor. Preferably, this assay is specific for specific calcium receptors. For example, assays that measure hormones whose production and secretion are regulated by specific inorganic ion receptor modulators are preferred.

Increases in [Ca ²⁺ ] ₁ use standard techniques such as using fluorometer indicators or measuring the increase in Cl ⁻ flow in Xenopus oocytes injected with nucleic acids encoding calcium receptors. Can be detected (see PCT / US93 / 01642 to Nemed et al. And WO94 / 18959). For example, poly (A) ⁺ mRNA may be used for cells expressing calcium receptors, such as epithelial cells, osteoclasts, glomerular kidney cells, myocardium renal cells, distal tubular kidney cells, decontamination and / or collection ducts. Thick ascending cells, epidermal keratinocytes, thyroid cells (C-cells), intestinal cells, central nervous system cells, peripheral nervous system cells, placental tropoblast, platelets, vascular smooth muscle cells, ventricular cells, gastrin secreting cells, Glucagon secreting cells, renal vessel membrane cells, breast cells, beta cells, adipocytes, immune cells and GI vascular cells. Preferably, the nucleic acid is obtained from epithelial cells, C-cells or osteoclasts. More preferably, the nucleic acid encodes a calcium receptor and is present on the plasma or vector.

Preferably, the molecule is calcium mimic and calcium degradable with an EC ₅₀ value or IC ₅₀ value of 5 μM or less, even more preferably 1 μM, 100 nmol, 10 nmol or 1 nmol or less. Such low EC ₅₀ values or IC ₅₀ values are advantageous because lower concentrations of molecules can be used for treatment or diagnosis in vivo or in vitro. The discovery of molecules with such low EC ₅₀ values or IC ₅₀ values allows the design and synthesis of another molecule with similar efficacy and effects.

In a preferred embodiment, the calcium receptor modulator inhibits the secretion of epithelial hormones from epithelial cells in vitro and reduces PTH secretion in vivo, stimulates the secretion of calcitonin from C-cells in vitro, and inhibits calcitonin in vivo. It is a calcium mimetic that raises levels or blocks osteoclast bone uptake in vitro and inhibits bone uptake in vivo.

In another preferred embodiment, the calcium receptor modulator is a calcium breakdown type that induces secretion of epithelial hormones from epithelial cells in vitro and elevates the levels of epithelial hormones in vivo.

Preferably, the agent selectively targets inorganic ion receptor activity, more preferably calcium receptor activity in certain cells. “Selectively” herein means that the molecule has a greater effect on inorganic ion receptor activity in one cell species than in another cell species for a given concentration of agent. Preferably, the differential effect is at least 10 times. Preferably the concentration relates to the plasma concentration of blood and the effect measured is the production of extracellular carriers such as plasma calcitonin, epithelial hormone or plasma calcium. For example, in a preferred embodiment, the agent selectively targets PTH secretion for calcitonin secretion.

In another preferred embodiment, epithelial cells, osteoclasts, glomerular kidney cells, mesothelial tubular kidney cells, distal tubular kidney cells, thickened ascending cells of Henle's and / or collecting ducts, central nervous system cells, peripheral nervous system cells, epidermis Keratinocytes, thyroid cells (C-cells), intestinal cells, placental tropoblast, platelets, vascular smooth muscle cells, ventricular cells, gastrin secretory cells, glucagon secreting cells, renal vessel membrane cells, breast cells, beta cells At least one molecule, but not all cells selected from the group consisting of adipocytes, immune cells and GI vascular cells, has an EC ₅₀ value or IC ₅₀ value of 5 μM or less.

Preferably, inorganic ion receptor modulators mimic or block all effects of extracellular ions in cells with inorganic ion receptors. For example, calcium receptor modulators preferably mimic or block all effects of extracellular ions in cells with calcium receptors. Calcium mimetics do not have to have all the biological activities of extracellular Ca ²⁺ , but rather at least one activity is mimicked. Similarly, the calcium degradation type does not need to reduce or inhibit all of the activity caused by extracellular calcium. In addition, different calcium mimetics and different calcium degradation forms do not need to bind to the same site on the calcium receptor as extracellular Ca ²⁺ binds to function.

A. Calcium Mimic

The ability of a molecule to mimic or block the activity of Ca ²⁺ at a calcium receptor can be measured using methods known in the art, including Nem et al. PCT / US93 / 01642 and WO / 18959. It is disclosed in the call. For example, calcium mimetics have one or more of the following activities, preferably all of the following when tested for epithelial cells in vitro:

1. The molecule causes a rapid (peaking time: less than 5 seconds) transient increase in [Ca ²⁺ ] ₁ that does not respond to inhibition of 1 μM La ³⁺ or 1 μM Gd ³⁺ . [Ca ^2+] increases when the other _one is a Ca ²⁺ is not present cell ionomycin (in the case other cell Ca ²⁺ is not present) is eliminated by pre-treatment.

2. The molecule enhances the increase in [Ca ²⁺ ] ₁ caused by sub-maximal concentrations of extracellular Ca ²⁺ .

3. The increase in [Ca ²⁺ ] ₁ caused by extracellular Ca ²⁺ is not inhibited by dihydropyridine.

4. The transient increase in [Ca ²⁺ ] ₁ induced by the molecule is eliminated by pretreatment with 10 mM sodium fluoride for 10 minutes.

5. Temporary increase in [Ca ²⁺ ] ₁ induced by the molecule was pretreated with an activator of protein kinase C (PKC) such as phorbol myristate acetate (PMA), mazelein or (-)-indolaccam V. By reducing. The overall effect of the protein kinase C activator is to shift the concentration-response curve of the molecule to the right without affecting maximal response.

6. The molecule causes a rapid (less than 30 seconds) increase in the formation of inositol-1,4,5-triphosphate and / or diacylglycerol.

7. The molecule inhibits cyclic AMP formation stimulated with dopamine- or isoproterenol.

8. The molecule inhibits PTH secretion.

9. Pretreatment with Pertussis toxin (greater than 4 hours at 100 ng / ml) blocks the inhibitory effect of the molecule on cyclic AMP formation, but increases [Ca ²⁺ ] _l , inositol-1, It does not affect the formation of 4,5-triphosphate or diacylglycerol and does not lower PTH secretion.

10. The molecule induces an increase in Cl ⁻ flow in xenopus orthosite injected with Fli (A) ⁺ rich mRNA taken from bovine or human epithelial cells, but infused with water or rat brain or liver mRNA It has no effect on xenopus orthosite.

11. Similarly, using cloned calcium receptors taken from epithelial cells, the molecule will react at xenofus orthosite injected with a purified cDNA or mRNA that encodes a tax.

Other calcium activities can be measured using the available techniques disclosed in PCT / US93 / 01642 to Nemed et al. And WO94 / 18959. Dynamic definitions of molecules that mimic Ca ²⁺ activity in other calcium reactive cells, preferably calcium receptors, are provided herein and in Ned et al., PCT / US93 / 01642 and WO94 / 18959. Obvious from the examples.

Preferably, the agent as determined by the bioassay disclosed herein or as determined by PCT / US93 / 01642 and Neh. WO94 / 18959, such as Nemed, is characterized by the following activities, ie less than 30 seconds. period causes a transient increase in internal calcium during and (preferably by mobilizing internal calcium), sikimyeo caused a rapid increase in [Ca ^2+] ₁ in 30 seconds or less, a ₁ [Ca ^2+] (for more than 30 seconds) Cause a sustained increase (preferably by introducing external calcium), preferably causing an increase in inositol-1,4,5-triphosphate or diacylglycerol levels in less than 60 seconds, and dopamine- or isoprotere It has at least one activity, more preferably all of these activities, which inhibits cyclic AMP formation stimulated with knol.

The temporary increase in [Ca ²⁺ ] ₁ is preferably eliminated by pretreatment of the cells for 10 minutes with 10 mM sodium fluoride or such temporary increase is achieved with phorbol myristate acetate (PMA), mazelein or (-)-indolakcam. Reduced by simple pretreatment (less than 10 minutes) of cells with an activator of protein kinase C (PKC) such as V.

B. Calcium Degradation Type

The ability of a molecule to block the activity of external calcium can be measured using standard techniques and is disclosed in PCT / US93 / 01642 to Nem et al. And WO 94/18959. For example, the molecule blocks the effect of external calcium when used on epithelial cells, and when tested against epithelial cells in vitro, it has one or more of the following activities, preferably all of the following:

1. The molecule contains an increased concentration of extracellular Ca ²⁺ .

(a) increasing [Ca ²⁺ ] ₁ ,

(b) mobilize intracellular Ca ²⁺ ,

(c) increases the formation of inositol-1,4,5-triphosphate,

(d) decreases cyclic AMP formation stimulated with dopamine- or isoproterenol,

(e) inhibits PTH secretion

Block the ability partially or completely

2. The molecule blocks an increase in Cl ⁻ flow in xenopus orthosite injected with poly (A) ⁺ mRNA taken from bovine or human epithelial cells caused by extracellular Ca ²⁺ or calcium mimetic compounds. It is not effective in xenopus orthosite injected with water, liver or liver mRNA.

3. Similarly, using cloned calcium receptors taken from epithelial cells, the molecule injects specific cDNAs, mRNAs or cRNAs encoding calcium receptors induced by extracellular Ca ²⁺ or calcium mimetic compounds. It will block the reaction in one xenopus orthosite.

Comparable definitions of molecules that block Ca ²⁺ activity in calcium reactive cells, preferably calcium receptors, are provided herein and in the examples provided in PCT / US93 / 01642 to Nemed et al. And WO94 / 18959. It is obvious from.

III. Treatment of a disease or disorder

Preferred uses of the compounds disclosed by the present invention are in the treatment or prevention of different diseases or disorders by modulating inorganic ion receptor activity. Inorganic ion receptor modulators of the present invention act on inorganic ion receptors to produce one or more cellular effects, ultimately providing therapeutic effects.

Different diseases or conditions can be treated by targeting the cells with inorganic ion receptors such as calcium receptors by the present invention. For example, primary hyperthyroidism (HPT) is characterized by elevated levels of hypercalcium blood and circulating PTH. A defect associated with major types of HPT is that the sensitivity of epithelial cells to negative feedback regulation by extracellular Ca ²⁺ is reduced. Thus, in primary HPT tissue taken from the patient, "reference points (set-point)" for Ca ²⁺ other cells are moved to the right so as to have to high Ca ²⁺ concentration than in the other normal cells in suppressing PTH secretion. Moreover, in primary HPT, much higher concentrations of extracellular Ca ²⁺ often only partially inhibit PTH secretion. In secondary (uremic) HPTs, a similar increase in baseline for extracellular Ca ²⁺ is observed, even though the extent to which Ca ²⁺ inhibits PTH secretion is normal. Changes in PTH secretion are reference points for the [Ca ^2+] ₁ increase in [Ca ^2+], along with the _first change, caused by the Ca ²⁺ extracellular is moved to the right, increasing the size is reduced.

Molecules that mimic the action of extracellular Ca ²⁺ are beneficial for long term treatment of both primary and secondary HPTs. The molecule helps to alleviate the signs of hypercalcemia by adding the stimuli necessary to suppress PTH secretion that cannot occur only by signs of hypercalcemia. Molecules with greater efficiency than extracellular Ca ²⁺ can overcome the distinct non-inhibitory components of PTH secretion that are particularly problematic for adenomas. Alternatively or in addition, the molecule may inhibit the synthesis of PTH as prolonged hypercalcemia has been shown to inhibit the levels of preprePTH mRNA in bovine and human adenoma epithelial tissues. Prolonged hypercalcemia also also inhibits proliferation of epithelial cells in vitro, just as calcium mimetics can be effective in limiting the aberrant proliferative properties of secondary HPT.

Cells other than epithelial cells can respond directly to physiological changes in extracellular Ca ²⁺ concentrations. For example, calcitonin secretion from thyroid cells (C-cells) is regulated by changes in the concentration of extracellular Ca ²⁺ .

The isolated osteoclasts respond to increases in extracellular Ca ²⁺ concentration corresponding to the increase in [Ca ^2+] _l caused in part by the mobilization of intracellular Ca ²⁺ cells. An increase in [Ca ²⁺ ] ₁ in osteoclasts is associated with inhibition of bone resorption. The release of alkaline phosphatase from osteoblastic osteoclasts is directly stimulated by calcium.

Like PTH secretion, renin secretion from renal glomeruli cells is inhibited by increasing concentrations of extracellular Ca ²⁺ . Extracellular Ca ²⁺ causes the mobilization of intracellular Ca ²⁺ in these cells cell. In other kidney cells, elevated Ca ²⁺ inhibits the formation of 1,25 (OH) ₂ -vitamin D by mesial tubule cells and stimulates the production of calcium binding proteins in distal tubule cells. Inhibits vasopressin's action on tubal uptake of Ca ²⁺ and Mg ²⁺ and Henley's thick bone marrow ascending angle (MTAL), decreases vasopressin's action on cortical coronary cells, Responds to calcium as it affects vascular smooth muscle cells.

Calcium also promotes differentiation of intestinal earbled cells, breast cells and skin cells, inhibits ventricular natriuretic peptide secretion from the ventricles, lowers cAMP accumulation in platelets, alters gastrin and glucagon secretion, and vascular activity It acts on vascular smooth muscle cells to modify cell secretion of factors, and affects cells of the central and peripheral nervous systems.

Thus, in addition to the role of Ca ²⁺ as an intracellular signal everywhere, there are sufficient indications suggesting that Ca ²⁺ also acts as a cellular signal for regulating the response of certain differentiated cells. Molecules of the invention can be used in the treatment of diseases or disorders associated with disrupted Ca ²⁺ responses in such cells.

Certain diseases and conditions that can be treated or prevented based on the affected cells include central nervous system diseases such as sudden seizures, seizures, head injury, spinal cord injury, hypoxia-induced neurons such as cardiac arrest or neonatal distress. Neurodegenerative diseases such as injury, epilepsy, Alzheimer's disease, Huntington's disease and Parkinson's disease, dementia, muscle tension, depression, anxiety, fear, obsessive-compulsive disorder, post-traumatic stress disorder, schizophrenia, neuroleptic malignant syndrome, and Tourette syndrome ; Diseases associated with excessive water resorption by the kidney, such as inappropriate ADH secretion symptoms (SIAH), cirrhosis, heart attacks, and nephropathy; High blood pressure; Preventing and / or reducing renal toxicity from cationic antibiotics (eg aminoglycoside antibiotics); Digestive tract active diseases such as diarrhea and convulsive colon; GI ulcer disease; GI uptake disorders such as pyromatosis; And autoimmune diseases and organ transplant rejection.

Inorganic ion receptor modulators of the present invention are commonly used in the treatment of humans, but they are used in other primates, pigs, cattle, and poultry chickens, live animals such as sports animals, and other warm blood such as pets such as horses, dogs, and cats. It can also be used to treat the same or similar disease or condition in an animal.

IV. administration

Molecules of the invention can be formulated in a variety of dosage forms for treating patients by modulating inorganic ion receptor activity. Techniques and formulation methods for the administration of the compounds can generally be found in Remington's Pharmaceutical Science, Mack Publishing Co., Easton, PA. Ion mimics and ion dissociation are discussed in PCT / US93 / 01642 to Nemed et al. And WO 94/18959.

Suitable forms depend in part on the use or route of infusion, for example oral, transdermal or injectable. This form must allow the agent to reach the target cell regardless of whether the target cell is present in the multicellular host or culture. For example, the pharmacological agent or composition to be injected into the blood stream must be soluble at the concentration used. Other factors are known in the art and include considerations such as toxicity and form that prevent the formulation or composition from exerting its action.

The formulations may also be formulated with pharmaceutically acceptable salts such as acid addition salts and combinations thereof. Formulations of such salts may facilitate pharmacological use by changing the physical properties of the formulation without interfering with their physiological effects. Examples of useful alterations in physical properties include lowering the melting point to facilitate administration through the mucosa and increasing solubility to facilitate the administration of high concentrations of drugs.

For systemic administration, oral administration is preferred. Alternative injection methods that can be used are, for example, intramuscular, intravenous, intraperitoneal and subcutaneous administration. Injecting the molecules of the invention may be formulated with a physiologically compatible buffer such as a liquid solution, preferably Hank's solution or Ringer's solution. In addition, the molecules may be formulated in solid form and redissolved or suspended immediately prior to use. It may also be produced in lyophilized form.

Systemic administration may also be via mucosal or percutaneous means, and the molecule may be administered orally. For administration via the mucous membranes or by percutaneous means, the preparations may employ suitable penetrants for the barrier to be permeated. These penetrants are generally known in the art and include, for example, bile salts and fuzidic acid derivatives for administration through the mucosa. In addition, a cleaning agent may be used to facilitate permeation. Administration through the mucosa can be done, for example, via nasal spray or using suppositories. For oral administration, the molecules are formulated in conventional oral dosage forms such as capsules, tablets and tonics.

For topical administration, the molecules of the invention are formulated in ointments or slaves, gels or creams, as is generally known in the art.

In general, the therapeutically effective amount of the molecule is about 1 nmol to 3 μmol, preferably 0.1 nmol to 1 μmol, depending on its EC ₅₀ or IC ₅₀ and the age and body size of the patient and the patient's associated disease or condition. Generally the amount is about 0.1 to 50 mg, preferably 0.01 to 20 mg per kg animal to be treated.

V. Examples

The compounds disclosed herein can be synthesized using standard techniques as disclosed in PCT / US93 / 01642 to Nemed et al. And WO 94/18959. Examples describing the synthesis of compounds 4L, 8J, 8U, 9R, 11X, 12U, 12V, 12Z, 14U, 16M and 16P are provided below. Compounds 4L, 8J, 8U, 11X, and 16M were prepared by condensation of primary amines with aldehydes or ketones in the presence of titanium (IV) isopropoxide. The resulting intermediate imine was then reduced intact by the action of sodium cyanoborohydride, sodium borohydride or sodium triacetoxyborohydride. Intermediate enamines for the synthesis of compound 8U were catalytically reduced using palladium hydroxide.

Synthesis of compounds 9R, 14U and 16P was prepared by reductive amination of the primary amine with commercially available aldehydes or ketones in the presence of sodium cyanoborohydride or sodium triaceoxyborohydride. For the synthesis of these three compounds 9R, 14U and 16P, the use of sodium triacetoxyborohydride than the use of sodium cyanocyborohydride can yield the desired diastereomer with greater diastereoselectivity. Turned out to be. These isomer-rich mixtures were further purified into single diastereomers by conventional phase HPLC or by recrystallization from organic solvents.

Compounds 12U, 12V and 12Z were prepared by diisobutylaluminum hydride (DIBAL-H) mediated condensation of amines and nitriles. The resulting intermediate imine was reduced by reacting sodium cyanoborohydride or sodium borohydride intact. Intermediate alkenes (compounds 12U and 12V) were reduced by catalytic hydrogenation in EtOH using a palladium on carbon catalyst. The compound converted to the corresponding hydrochloride gave a white solid by treating the free base with etheric HCl.

The amines in this synthesis were purchased from (1) Aldrich Chemical Company, Milwaukee, WI or (2) Selgen Corporation, Warren, NJ, or (3) synthesized using standard techniques. All other reagents were purchased from Aldrich Chemicals.

Example 1 Synthesis of Compound 4L

N-3-phenyl-1-propyl-1- (1-naphthyl) ethylamine

135 mg (1 mmol) of 3-phenyl-1-propynamine, 170 mg (1 mmol) of 1'-acetonaphtone and 355 mg (1.3 mmol) of titanium (IV) isopropoxide were stirred at room temperature for 1 hour. The reaction was treated with 1 mL of 1 M ethanol sodium cyanoborohydride and stirred at rt for 16 h. The reaction was diluted with ether and treated with 0.1 mL of water. The reaction was centrifuged, the ether layer removed and concentrated to milky oil. 10 mg of this subfraction was purified by HPLC (phenomenex, 1.0 × 25 cm, 5 μΜ silica) using a gradient of dichloromethane to 10% methanol in dichloromethane containing 0.1% isopropylamine. This resulted in the product (free base) as a single component by GC / El-MS (R _t = 10.48 min) m / z (rel. Int.). 289 (M ⁺ , 11), 274 (63), 184 (5), 162 (5), 155 (100), 141 (18), 115 (8), 91 (45), 77 (5).

Example 2: Synthesis of Compound 8J

N- (3-phenylpropyl) -1- (3-thiomethylphenyl) ethylamine hydrochloride

2.7 g (20 mmol) of 3'-aminoacetophenone was dissolved in 4 mL concentrated HCl, 4 g ice and 8 mL water. The solution was cooled to 0 ° C. and 1.45 g (21 mmol) of sodium nitrite dissolved in 3 to 5 mL of water were added over 5 minutes while maintaining the temperature below 6 ° C. 1.75 g (25 mmol) of sodium thiomethoxide were dissolved in 5 mL of water and cooled to 0 ° C. To this solution was added diazonium salt over 10 minutes while maintaining the temperature below 10 ° C. The temperature was raised to ambient temperature with stirring for an additional hour. The reaction mixture was partitioned between ether and water. The ether layer was separated, washed with bicarbonate and sodium chloride and dried over sodium sulfate. The ether layer was evaporated to give 3'-thiomethylacetophenone in 74% yield. The crude material was purified by distillation under reduced pressure.

0.13 g (1 mmol) of 3-phenylpropylamine, 0.17 g (1 mmol) of 3'-thiomethylacetophenone and 0.36 g (1.25 mmol) of titanium (IV) isopropoxide were mixed together and allowed to stand for 4 hours. . To this was added 1 mL of ethanol and 0.063 g (1 mmol) of sodium cyanoborohydride and the reaction was stirred overnight. The reaction was treated by adding 4 mL of ether and 200 μL of water. The reaction was vortexed and then spun in a centrifuge to separate the solids. The ether layer was separated from the precipitate and the solvent was removed in vacuo. The oil was redissolved in dichloromethane and the compound was purified by preparative TLC on silica gel using 3% methanol / dichloromethanol eluent to afford the title compound as a pure oil: GC / EI-MS (R _t = 7.64 min) m / z (rel. int.) 285 (M ⁺ , 18), 270 (90), 180 (17), 151 (100), 136 (32), 104 (17), 91 (54), 77 (13) .

Example 3: Synthesis of Compound 8U

N-3- (2-methoxyphenyl) -1-propyl- (R) -3-methoxy-α-methylbenzylamine hydrochloride

3.02 g (20 mmol) of (R)-(+)-3-methoxy-α-methylbenzylamine, 3.24 g (20 mmol) of 2-methoxycinnaaldehyde and 8.53 g (30) of titanium (IV) isopropoxide mmol, 1.5 equiv) was stirred at rt for 2 h and treated with 20 mL of 1 M ethanol sodium cyanoborohydride. The reaction was stirred overnight (16 hours), diluted with diethyl ether and treated with 1.44 mL (80 mmol, 4 equiv) of water. After this was mixed for 1 hour, the reaction mixture was centrifuged, the ether layer was removed and concentrated to an oil. This material was dissolved in glacial acetic acid, shaken with palladium hydroxide and hydrogenated under 60 psi hydrogen for 2 hours at room temperature. The catalyst was filtered off and the resulting solution was concentrated to a thick oil. This material was dissolved in dichloromethane and neutralized with 1N NaOH. The dichloromethane solution was separated from the aqueous phase, dried over anhydrous calcium carbonate and concentrated to an oil. This material was dissolved in ether and treated with 1 M HCl in diethyl ether. The resulting precipitate (white solid) was collected, washed with diethyl ether and airflow dried. A single component emerged from GC / El-MS (R _t = 9.69 min) of this material (free base): m / z (rel. Int.) 299 (M ⁺ , 21), 284 (100), 164 (17 ), 150 (8), 135 (81), 121 (40), 102 (17), 91 (43), 77 (18).

Example 4: Synthesis of Compound 9R

(R) -N- (1- (2-naphthyl) ethyl)-(R) -1- (1-naphthyl) ethylamine hydrochloride

10.0 g (58 mmol) of (R)-(+)-1- (1-naphthyl) ethylamine, 9.4 g (56 mmol) of 2'-acetonaprone and 20.7 g (73.0) of titanium (IV) isopropoxide mmol) and ethanol (anhydrous) were heated to 60 ° C. for 3 h. Then, 3.67 g (58.4 mmol) of sodium cyanoborohydride (NaCNBH ₃ ) were added. To this reaction mixture was added 1 liter of ether and 10 mL of water, and the resulting precipitate was removed by centrifugation. The supernatant was evaporated under vacuum and the crude product was recrystallized four times from hot hexanes to give 1.5 g of diastereomer of pure (98 +%) purity. The free base was dissolved in hexane, filtered and then etheric HCl was added to precipitate the product as a white solid (1.1 g, 6% yield).

Melting point: softened at 200-240 ° C. (decomposition).

Example 5: Synthesis of Compound 11X

N- (4-isopropylbenzyl)-(R) -1- (1-naphthyl) ethylamine hydrochloride

1.06 g (6.2 mmol) of (R)-(+)-1- (1-naphthyl) ethylamine, 0.92 g (6.2 mmol) of 4-isopropylbenzaldehyde and 2.2 g (7.7 mmol) of titanium (IV) isopropoxide ) Was heated to 100 ° C. for 5 minutes and then stirred at room temperature for 4 hours. Then 0.39 g (6.2 mmol) of sodium cyanoborohydride (NaCNBH ₃ ) were added followed by 1 mL of ethanol. The reaction mixture was stirred at rt for 18 h. 100 mL of ether and 1 mL of water were added to the reaction mixture, and the resulting precipitate was removed by centrifugation. The supernatant was evaporated under vacuum and the crude product was chromatographed (eluted with 1% MeOH / CHCl ₃ ) on silica gel (50 mm × 30 cm column). The chromatographed material was then dissolved in hexane and ethereal HCl was added to precipitate the product as a white solid (0.67 g, 35% yield). Melting Point: 257-259 ℃

Example 6: Synthesis of Compound 12U

N-3- (2-methylphenyl) -1-propyl- (R) -3-methoxy-α-methylbenzylamine hydrochloride

A solution of 1.43 g (10 mmol) of 2-methylcinnamonitrile in 10 mL of dichloromethane was cooled to 0 ° C. and treated dropwise with 10 mL of 1 M diisobutylaluminum hydride (dichloromethane) for 15 minutes. The reaction was stirred at 0 ° C. for 15 minutes and treated dropwise for 15 minutes with a solution of 1.51 g (10 mmol) of 1 M (R)-(+)-1-3-methoxy-α-methylbenzylamine. The reaction was stirred at 0 ° C. for 1 hour and poured into 100 mL of an ethanol solution containing 1 g (16 mmol) of sodium cyanoborohydride. The reaction mixture was stirred at rt for 48 h. The reaction mixture was diluted with ether and neutralized with 1N NaOH. The ether layer was removed, dried over anhydrous potassium carbonate and concentrated to an oil. This material was chromatographed over silica using a gradient of dichloromethane to 5% methanol in dichloromethane to GC / El-MS (R _t = 10.06 min) m / z (rel. Int.) As a single component. Unsaturated intermediates were obtained. 281 (M ⁺ , 17), 266 (59), 176 (19), 146 (65), 135 (73), 131 (100), 91 (21), 77 (13).

The unsaturated intermediate in ethanol was hydrogenated in the presence of a palladium on carbon catalyst for 16 hours at room temperature (1 atm H ₂ ). The product from this reaction was converted to hydrochloride by treatment with 1 M HCl in diethyl ether. GC / El-MS (R _t = 9.31 min) of this material (free base) showed a single component: m / z (rel. Int.) 283 (M ⁺ , 21), 268 (100), 164 (12 ), 148 (8), 135 (85), 121 (12), 105 (49). 91 (23), 77 (21).

Example 7: Synthesis of Compound 12V

N-3- (3-methylphenyl) -1-propyl- (R) -3-methoxy-α-methylbenzylamine hydrochloride

The compound was prepared following the procedure of Example 6 except for using 2-methylcinnamonitrile. The unsaturated intermediate was a single component as a result of GC / El-MS (R _t = 10.21 min) m / z (rel. Int.). 281 (M ⁺ , 57), 266 (86), 146 (98), 135 (88), 131 (100), 115 (43), 102 (26), 91 (43), 77 (18). This material was reduced and hydrochloride was formed using the procedure described in Example 6 to yield the product. GC / El-MS (R _t = 9.18 min) of this material (free base) showed a single component: m / z (rel. Int.) 283 (M ⁺ , 19), 268 (100), 164 ( 11), 148 (8), 135 (76), 121 (16), 105 (45), 91 (23), 77 (21).

Example 8: Synthesis of Compound 12Z

N-3- (2-chlorophenyl) -1-propyl- (R) -1- (1-naphthyl) ethylamine hydrochloride

The compound was prepared following the procedure of Example 6 except that 2-chlorohydrocinnamonitrile and (R)-(+)-1- (1-naphthyl) ethylamine were used in an amount of 10 mmol. This material was chromatographed over silica using a gradient of dichloromethane to 5% methanol in dichloromethane and the product was obtained as a single component by TLC analysis (5% methanol in dichloromethane). This was treated with 1 M HCl in diethyl ether to prepare hydrochloride.

Example 9: Synthesis of Compound 14U

(R) -N- (1- (1-methoxyphenyl) ethyl)-(R) -1- (1-naphthyl) ethylamine hydrochloride

1.1 g (6.2 mmol) of (R)-(+)-1- (1-naphthyl) ethylamine, 0.93 g (6.2 mmol) of 4'-methoxyacetophenone and 2.2 g of titanium (IV) isopropoxide 7.7 mmol) and 1 mL of ethanol (anhydrous) were heated to 60 ° C. for 3 hours. Then 0.39 g (6.2 mmol) of sodium cyanoborohydride (NaCNBH ₃ ) were added and the reaction mixture was stirred for 18 hours at room temperature. 200 mL of ether and 2 mL of water were added to the reaction mixture, and the resulting precipitate was removed by centrifugation. The supernatant was evaporated under vacuum and the crude product was chromatographed (eluted with 1% MeOH / CHCl ₃ ) on silica gel (25 mm × 25 cm column). This material subfraction was subjected to HPLC chromatography [SELECTOSIL, 5 μΜ silica gel; 25 cm x 10.0 mm (Phenomenex, Torrance, CA), 4 mL / min; UV det. 275 nM; 12% ethyl acetate-88% hexanes (elution time 12.0 min)]. The diastereomer purified by HPLC was then dissolved in hexanes and etheric HCl was added to precipitate the product as a white solid (20 mg). Melting point: 209-210 ° C. (decomposition).

Example 10 Synthesis of Compound 16M

N- (3-chloro-4-methoxybenzyl)-(R) -1- (1-naphthyl) ethylamine hydrochloride

6.6 g (39 mmol) of (R)-(+)-1- (1-naphthyl) ethylamine, 6.6 g (39 mmol) of 3'-chloro-4'-methoxybenzaldehyde and isopropoxide of titanium (IV) Seed 13.8 g (48.8 mmol) and 30 mL of ethanol (anhydrous) were heated to 80 ° C. for 30 minutes and then stirred at room temperature for 3 hours. Then, 2.45 g (39 mmol) of sodium cyanoborohydride (NaCNBH ₃ ) were added. The reaction mixture was stirred at rt for 18 h. 100 mL of ether and 2 mL of water were added to the reaction mixture, and the resulting precipitate was removed by centrifugation. The supernatant was evaporated under vacuum and the crude product was chromatographed (eluted with CH ₂ Cl ₂ ) on silica gel (50 mm × 30 cm column). The chromatographed material was then dissolved in 500 mL of hexane and 0.2 μM Norit Filter off with bleaching and then add etheric HCl to precipitate the product as a white solid (10.2 g, 56% yield). Melting point: 241-242 ° C. (decomposition).

Example 11: Synthesis of Compound 16P

4-methoxy-3-methylacetophenone [16P precursor]

5.0 g (33.3 mmol) of 4'-hydroxy-3'-methylacetophenone, 5.7 g (40.0 mmol) of iodomethane, 23.0 g (167 mmol) of K ₂ CO ₃ (granular, anhydrous), and 250 mL of acetone It was refluxed for 3 hours. The reaction mixture was then cooled to room temperature, filtered to remove inorganic salts and evaporated in vacuo. The crude product was dissolved in 100 mL of ether and washed with water (2 × 20 mL). The organic layer was dried over Na ₂ SO ₄ and evaporated to give 4.5 g of product, yield 82.4%. The ketone was used without further purification in the next reaction.

(R) -N- (1- (4-methoxy-3-methylphenyl) ethyl)-(R) -1- (1-naphthyl) ethylamine hydrochloride [Compound 16P]

4.24 g (24.8 mmol) of (R)-(+)-1- (1-naphthyl) ethylamine, 4.06 g (24.8 mmol) of 4'-methoxy-3'-methylacetophenone and titanium (IV) isopro 8.8 g (30.9 mmol) of oxide and 1 mL of ethanol (anhydrous) were heated to 100 ° C. for 2 hours. 45 mL of isopropane was added thereto, and the reaction was cooled to 10 ° C. in an ice bath. Then 10.5 g (49.5 mmol) of sodium triacetoxyborohydride (NaHB (O ₂ CCH ₃ ) ₃ ) was added in several portions over 15 minutes. The reaction mixture was heated to 70 ° C. for 18 hours. The mixture was cooled to room temperature and poured into 400 mL of ether. The suspension was centrifuged, the supernatant collected and the pellet washed with 400 mL of ether. The combined organic washes were evaporated under vacuum. The residue was dissolved in 400 mL of ether and washed with 1 N NaOH (4 × 50 mL) and water (2 × 50 mL). The organic layer was dried over Na ₂ S0 ₄ , filtered and evaporated in vacuo. Anhydrous ethanol was added to the moist residue, followed by thorough evaporation using a rotary evaporator to yield an oil. This mixture was then chromatographed on silica gel (50 mm × 30 cm) (eluted with 1% MeOH: 1% IPA: CHCl ₃ ) to give 4.8 g of oil.

The desired diastereomer was HPLC chromatography [SUPELCOSIL ^™ PLC-Si, 18 μΜ silica gel: 25 cm × 21.2 mm (Supelco, Bellefonte, PA), 7 mL / min; UV det. 275 nM; 20% ethyl acetate-80% hexanes (elution time 9.5-11.0 min)]. This mixture (100 mg / mL solution in eluent) was sprayed (800 μL droplets) to give 65 mg of the desired isomer. 1.0 g of pure material was obtained from multiple HPLC injections. The HPLC chromatographed material was dissolved in 50 mL of hexane and the hydrochloride was precipitated with etheric HCl. This salt was collected on fritted glass and washed with hexane to give a white solid (1.0 g). Melting point: 204-205 ° C. (decomposition).

Other aspects are included in the appended claims.

Claims (7)

Compounds of the formula or pharmaceutically acceptable salts thereof. In the above formula, Each X is independently selected from the group consisting of isopropyl, CH ₃ O, CH ₃ S and fused aromatic rings, m is 0-5. Compounds of the formula or pharmaceutically acceptable salts thereof. In the above formula, X is each independently CH ₃ , CH ₃ O, CH ₃ CH ₂ O, methylene dioxy, Br, Cl, F, CF ₃ , CH ₃ S, OH, CH ₃ CH ₂ , propyl, isopropyl, butyl, iso Butyl, t-butyl and fused aromatic rings; R is selected from the group consisting of hydrogen and methyl, m is 0-5. The compound of claim 2, or a pharmaceutically acceptable salt thereof, wherein R is H. 4. The compound or pharmaceutically acceptable salt thereof according to claim 3, wherein each X is independently selected from the group consisting of isopropyl, CH ₃ O, CH ₃ S and fused aromatic rings. Compounds of the formula or pharmaceutically acceptable salts thereof. A compound of the formula or pharmaceutically acceptable salt thereof, A disease selected from the group consisting of primary or secondary epithelial hyperplasia, Paget's disease, hypercalcemia, osteoporosis and hypertension, comprising the compound of any one of claims 1 to 6 and a pharmaceutically acceptable carrier or Pharmaceutical compositions for the treatment of diseases.