METHODS FOR INCREASING NEUROGENESIS

FIELD OF THE INVENTION

The present invention relates to methods and compounds useful for increasing neurogenesis. Methods and compounds for increasing dopaminergic differentiation are also provided. Methods for treating neurological disorders by increasing neurogenesis are also provided.

BACKGROUND OF THE INVENTION

Neurogenesis is the process by which neurons are created. Until recently, neurogenesis was thought to occur only in developing organisms, but evidence now supports the occurance of adult neurogenesis.

Neurogenesis is an important aspect of neuronal plasticity, enabling organisms to adapt to environmental changes and influencing learning and memory throughout life.

The process of adult neurogenesis includes neural stem cell division (i.e., cell proliferation), migration, maturation, and differentiation. Within the adult brain, the majority of new neurons arise from the subventricular zone (SVZ) of the lateral ventricles and the subgranular zone (SGZ) of the hippocampus, and are incorporated into the functional networks of both the olfactory bulb and the hippocampal dentate gyrus, suggesting significant roles for adult neurogenesis in brain function. Regulation of neurogenesis occurs at all levels including the proliferation, differentiation, and fate determination of adult neural progenitor cells (NPCs), and the survival, maturation, and integration of newborn neurons. Furthermore, these cells may be required for certain forms of learning and memory.

The ability to expand neural progenitor cells and manipulate their cell fate has important implications for numerous neurological disorders where specific cell types have been lost. Providing a supply of new and functional neural cells, including new neurons, has been proposed as a means to provide neurogenesis in order to ameliorate neural degeneration. A loss of neurogenesis and decrease in hippocampal volume has been associated with chronic stress and depression, and hippocampal size is decreased in patients with major depressive disorder, post-traumatic stress disorder (PTSD) and Cushing's disease. (See, e.g., Bremner et al. (1995) Am J Psychiatry 152:973-981; Sheline et al. (1996) Proc Natl Acad Sci USA 93:3908-3913.) Neurogenesis is required for the actions of antidepressants, and adult neurogenesis may counter the pathological effects of stress and chronic depression. (Santarelli et al. (2003) Science 301: 805-809.) Studies have also suggested that adult hippocampal neurogenesis may relieve symptoms in psychiatric and neurologic disorders such as addiction, epilepsy, and schizophrenia. (See, e.g., Eisch et al. (2008) J Neurosci 28:11785-11791; Warner-Schmidt and Duman (2006) Hippocampus 16:239-249.) Various strategies directed at providing neurogenesis have been proposed. For example, one strategy would be to transplant neural progenitor cells into the brain or spinal cord, such that the neural progenitor cells develop into specialized neural cells. Another strategy would be to increase or enhance endogenous neurogenesis by delivering biologically active molecules to the brain or spinal cord in order to stimulate the proliferation, migration, and differentiation of endogenous neural progenitor cells.

The availability of effective therapies for increasing or enhancing neurogenesis, however, is limited, and there exists a need for identifying compounds and methods for modulating neurogenesis. The present invention meets this need by providing methods and compounds effective at increasing neurogenesis and increasing neural differentiation and, in particular, dopaminergic differentiation.

SUMMARY OF THE INVENTION

The present invention provides methods and compounds useful for increasing neurogenesis. In some embodiments, the present invention provides a method for increasing neurogenesis, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing neurogenesis. The present invention also provides compounds for use in manufacturing a medicament for increasing neurogenesis, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme. In certain aspects, increasing neurogenesis is associated with increasing neural cell differentiation, increasing dopaminergic differentiation of neural cells, and increasing neural cell proliferation. In other aspects, the neural cell in these methods is a neural stem cell, a neural progenitor cell, or a neural precursor cell.

Subjects in need of such treatment include subjects having cognitive impairement, due to injury, to disease, or to other condition, or subjects having altered dopaminergic responses. Additional subjects suitable for treatment with increased neurogenesis include subjects having psychiatric disorders, such as addiction or schizophrenia, or subjects having a neurogenerative disorder, including Parkinson's disease, dystonia, essential tremor, or a chronic neurological disorder such as epilepsy. It is also contemplated that the present methods can be applied to treatment of subjects experiencing depression or stress. In one embodiment, the present invention provides methods for treating a subject with altered dopaminergic response, the method comprising administering to the subject a compound that inhibits HIF prolyl hydroxylase activity. The invention also provides for use of a compound that inhibits HIF prolyl hydroxlase activity in manufacturing a medicament for treatment of a subject having altered dopaminergic response.

The present invention provides methods for increasing neurogenesis in a subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HEF prolyl hydroxylase activity. In particular embodiments, the present invention provides methods for increasing dopaminergic neurogenesis in a subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HIF prolyl hydroxylase activity. The present invention also encompasses the use of a compound that inhibits HIF prolyl hydroxylase activity in manufacturing a medicament for increasing dopaminergic neurogenesis.

The present invention provides methods of treating a neurological disorder by increasing neurogenesis in a subject in need, the method comprising administering a compound that inhibits HIF prolyl hydroxylase activity to the subject, thereby increasing neurogenesis and treating the disorder. In some embodiments, the methods increase dopaminergic neurogenesis, thereby treating the disorder. In one embodiment, the invention provides methods of treating a neurological disorder by increasing neurogenesis in a subject in need, the method comprising administering a compound that inhibits HIF prolyl hydroxylase activity to the subject, wherein the neurological disorder is a neurodegenerative disorder. In particular embodiments, the invention provides methods of treating a neurological disorder by increasing neurogenesis in a subject in need, the method comprising administering a compound that inhibits HIF prolyl hydroxylase activity to the subject, wherein the neurological disorder is Parkinson's disease. In another embodiment, the invention provides methods of treating a neurological disorder by increasing neurogenesis in a subject in need, the method comprising administering a compound that inhibits HIF prolyl hydroxylase activity to the subject, wherein the neurological disorder is a chronic stress disorder. In another embodiment, the invention provides methods of treating a neurological disorder by increasing neurogenesis in a subject in need, the method comprising administering a compound that inhibits HIF prolyl hydroxylase activity to the subject, wherein the neurological disorder is a cognitive impairment.

The present invention further provides a method for increasing neural cell differentiation, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing neural cell differentiation. In particular embodiments, the present invention provides methods for increasing neural cell differentiation in a subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HIF prolyl hydroxylase activity. The present invention also provides compounds for use in manufacturing a medicament for increasing neural cell differentiation, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme. In certain embodiments, the neural cell in these methods is a neural progenitor cell such as, for example, a mesencephalic neural progenitor cell or a cortical (i.e., frontal) neural progenitor cell. In other embodiments, the neural cell in these methods is a neural stem cell, a neural progenitor cell, or a neural precursor cell.

In another embodiment, the present invention provides methods for increasing dopaminergic differentiation of neural cells, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing dopaminergic differentiation of the neural cell. In some embodiments, the present invention provides methods for increasing dopaminergic differentiation of neural cells in a subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HIF prolyl hydroxylase activity. In other embodiments, the present invention provides methods for increasing dopaminergic neurons (i.e., increasing the number of dopaminergic neurons), the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing dopaminergic neurons. The present invention also provides compounds for use in manufacturing a medicament for increasing neural dopaminergic differentiation, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme.

In another embodiment, the present invention provides a method for increasing neural cell proliferation, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing neural cell proliferation. The present invention also provides compounds for use in manufacturing a medicament for increasing neural cell proliferation, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme. In certain embodiments, the neural cell in these methods is a neural progenitor cell such as, for example, a mesencephalic neural progenitor cell or a cortical (i.e., frontal) neural progenitor cell. In yet other embodiments, the neural cell in these methods is a neural stem cell, a neural progenitor cell, or a neural precursor cell.

The compounds, methods, and medicaments of the present invention can be used to treat subjects wherein increased neurogenesis would be beneficial, in particular wherein increased hippocampal neurogenesis would be beneficial. Such subjects may include patients with chronic stress disorders or depression. In one embodiment, the compounds and methods can be used to treat subjects having or at risk for having a major depressive disorder, post-traumatic stress disorder (PTSD), or Cushing's disease. In another embodiment, the compounds and methods can be used to relieve symptoms in a subject having or at risk for having a psychiatric or neurologic disorder such as addiction, epilepsy, or schizophrenia. In another embodiment, the compounds and methods may be used to treat a subject with mild cognitive impairment (MCI) or dementia, e.g., age-related dementia. In one embodiment, the present invention provides methods for increasing neurogenesis in a subject having cognitive impairment, e.g., mild cognitive impairment, the method comprising administering to the subject a compound that inhibits HIF prolyl hydroxylase activity. In another embodiment, the invention provides for use of a compound that inhibits HIF prolyl hydroxylase activity in manufacturing a medicament for treatment of mild cognitive impairment. Increased neurogenesis, particularly increased dopaminergic neurogenesis, may also be beneficial in treatment and functional recovery in a subject having progressive neurological disorders including various neurodegenerative disorders, such as Parkinson's disease and essential tremor. In one embodiment, the present invention provides methods for increasing dopaminergic neurogenesis in a subject having a neurodegenerative disease, the method comprising administering to the subject a compound that inhibits

HEF prolyl hydroxylase activity. In one embodiment, the present invention provides methods for increasing dopaminergic neurogenesis in a subject having Parkinson's disease, the method comprising administering to the subject a compound that inhibits HEF prolyl hydroxylase activity. In one embodiment, the present invention provides methods for increasing dopaminergic neurogenesis in a subject having essential tremor, the method comprising administering to the subject a compound that inhibits HEF prolyl hydroxylase activity. In another embodiment, the invention provides for use of a compound that inhibits HEF prolyl hydroxylase activity in manufacturing a medicament for treatment of neurodegnerative disease such as Parkinson's disease, essential tremor, and various types of kinetic tremors.

In various embodiments, a compound used in the present methods is a structural mimetic of 2- oxoglutarate. In certain embodiments, the compound is a structural mimetic of 2-oxoglutarate that inhibits HEF prolyl hydroxylase activity competitively with respect to 2-oxoglutarate. In particular embodiments, compounds used in the present methods and medicaments provided herein are structural mimetics of 2-oxoglutarate, wherein the compound inhibits the target HEF prolyl hydroxylase enzyme competitively with respect to 2-oxoglutarate and noncompetitively with respect to iron.

In particular embodiments, compounds for use in the present invention include cyclic carboxamides, wherein the cyclic group is a carbocycle or a heterocycle. Therefore, in certain embodiments, the compounds used are carbocyclic carboxamides or heterocyclic carboxamides. In other embodiments, carbocyclic carboxamides for use in the present invention are naphthalene carboxamides. En yet other embodiments, heterocyclic carboxamides for use in the present invention are isoquinoline carboxamides and pyrrolopyridazine carboxamides.

Heterocyclic carboxamides for use in the present invention include hydroxy isoquinoline carboxamides, oxo pyrrolopyridazine carboxamides, hydroxy pyrrolopyridazine carboxamides, and hydroxy oxo pyrrolopyridazine carboxamides.

In other embodiments, compounds for use in the present invention include variously substituted 4- hydroxy-isoquinoline-3-carbonyl glycines, 2-oxo-pyrrolopyridazine-3-carbonyl glycines, 4-hydroxy- pyrrolopyridazine-3-carbonyl glycines, and 4-hydroxy-2-oxo-pyrrolopyridazine-3-carbonyl glycines. In particular embodiments, the compound used in the present invention is selected from the group consisting of [(4-Hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino] -acetic acid (Compound A), [(4-Hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (Compound B), {[l-Cyano-6-(2,6- dimethyl-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino} -acetic acid (Compound C), or {[4- Hydroxy-2 -oxo-1 -(4-trifluoromethyl-benzyl)-l,2-dihydro-pyrrolo[l,2-b]pyrida zine-3-carbonyl]-amino}- acetic acid (Compound D).

It is further contemplated that a compound for use in the present invention is a compound encompassed by one of Formulae I, Ia, Ib, Ic, and Id; Formula II; Formulae HI and IHa; Formulae IVA, IVB, IVC, and IVD; Formulae V, VA, VB, VC, and VD; Formula VI; Formula VII; Formula VIII; Formula DC; Formula

X; and Formula XI. Each of these formulae are detailed, infra.

These and other embodiments of the present invention will readily occur to those of skill in the art in light of the disclosure herein, and all such embodiments are specifically contemplated.

Each of the limitations of the invention can encompass various embodiments of the invention. It is, therefore, anticipated that each of the limitations of the invention involving any one element or combinations of elements can be included in each aspect of the invention. This invention is not limited in its application to the details of construction and the arrangement of components set forth in the following description or illustrated in the drawings. The invention is capable of other embodiments and of being practiced or of being carried out in various ways. Also, the phraseology and terminology used herein is for the purpose of description and should not be regarded as limiting. The use of "including," "comprising," or "having," "containing", "involving", and variations thereof herein, is meant to encompass the items listed thereafter and equivalents thereof as well as additional items. It must be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless context clearly dictates otherwise.

BRIEF DESCRIPTION OF THE DRAWINGS

Figures IA, IB, 1C, ID, and IE set forth data showing the effect of compound of the present invention, DFO, CPX, CoCl ₂ , and DMOG, respectively, on human neural progenitor cell viability.

Figure 2 sets for data showing compound of the present invention increased the number of human neural progenitor cells.

Figure 3 sets forth data showing compound of the present invention increased S-phase of cell cycle in human neural progenitor cells. Figures 4A, 4B, and 4C set forth data showing compound of the present invention increased neural cell proliferation and neurogenesis in human neural progenitor cells.

Figure 5 sets forth data showing compound of the present invention increased neural cell proliferation and neurogenesis in human neural progenitor cells.

Figure 6 sets forth data showing compound of the present invention increased dopaminergic differentiation in human neural progenitor cells.

Figure 7 sets forth data showing compound of the present invention increased dopaminergic differentiation in human neural progenitor cells.

Figure 8 set forth data showing compound of the present invention increased neuron specific enolase and tyrosine hydroxylase protein levels in human neural progenitor cells.

Figures 9A and 9B set forth data showing compound of the present invention increased percent neurospheres with neurites in neural progenitor cells.

Figures 1OA and IB set forth data showing compound of the present invention increased the number of neurites in neural progenitor cells.

Figure 11 sets forth data showing compound of the present invention increased VEGF mRNA expression in neural progenitor cells.

Figures 12A and 12B set forth data showing compound of the present invention increased the number of new mature neurons and improved contextual fear conditioning in an animal model.

DESCRIPTION OF THE INVENTION Before the present compositions and methods are described, it is to be understood that the invention is not limited to the particular methodologies, protocols, cell lines, assays, and reagents described, as these may vary. It is also to be understood that the terminology used herein is intended to describe particular embodiments of the present invention, and is in no way intended to limit the scope of the present invention as set forth in the appended claims. It must be noted that as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless context clearly dictates otherwise. Thus, for example, a reference to "a HIF-specifϊc 2-oxoglutarate dioxygenase enzyme" may include a plurality of such enzymes; a reference to a "compound that inhibits the activity of a hypoxia-inducible factor prolyl hydroxylase enzyme" may be a reference to one or more compounds that inhibits the activity of a hypoxia-inducible factor prolyl hydroxylase enzyme, and so forth.

Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices, and materials are now described. All publications cited herein are incorporated herein by reference in their entirety for the purpose of describing and disclosing the methodologies, reagents, and tools reported in the publications that might be used in connection with the invention. Nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.

The practice of the present invention will employ, unless otherwise indicated, conventional methods of chemistry, biochemistry, molecular biology, cell biology, genetics, immunology and pharmacology, within the skill of the art. Such techniques are explained fully in the literature. See, e.g., Gennaro, A.R., ed. (1990) Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing Co.; Hardman, J.G.,

Limbird, L.E., and Gilman, A.G., eds. (2001) The Pharmacological Basis of Therapeutics, 10th ed., McGraw-Hill Co.; Colowick, S. et al., eds., Methods In Enzymology, Academic Press, Inc.; Weir, D.M., and Blackwell, CC, eds. (1986) Handbook of Experimental Immunology, VoIs. I-IV, Blackwell Scientific Publications; Maniatis, T. et al., eds. (1989) Molecular Cloning: A Laboratory Manual, 2nd edition, VoIs. I-III, Cold Spring Harbor Laboratory Press; Ausubel, F.M. et al., eds. (1999) Short

Protocols in Molecular Biology, 4th edition, John Wiley & Sons; Ream et al., eds. (1998) Molecular Biology Techniques: An Intensive Laboratory Course, Academic Press; Newton, C.R., and Graham, A., eds. (1997) PCR (Introduction to Biotechniques Series), 2nd ed., Springer Verlag.

The section headings are used herein for organizational purposes only, and are not to be construed as in any way limiting the subject matter described herein.

Methods

The present invention relates to the discovery that inhibiting hypoxia-inducible factor (HIF) hydroxylase is effective at increasing neurogenesis. In certain aspects, the present invention provides methods and compounds useful for increasing neurogenesis. In one embodiment, the present invention provides a method for increasing neurogenesis, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing neurogenesis. In particular embodiments, the present invention provides methods for increasing neurogenesis in a subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HIF prolyl hydroxylase activity. The present invention also provides compounds for use in manufacturing a medicament for increasing neurogenesis, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme. In certain aspects, increasing neurogenesis is associated with increasing neural cell differentiation, increasing dopaminergic differentiation of neural cells, and increasing neural cell proliferation. In other aspects, the neural cell in these methods is a neural stem cell, a neural progenitor cell, or a neural precursor cell. In various embodiments, a compound that inhibits HIF prolyl hydroxylase activity is a cyclic carboxamide. In particular embodiments, the compound for use in the present invention is selected from a carbocyclic carboxamide or a heterocyclic carboxamide. In certain embodiments, a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme is a heterocyclic carbonyl glycine of Formula VI.

In one aspect, the present invention provides a method for increasing dopaminergic neurogenesis in subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HIF prolyl hydroxylase activity. The present invention also provides for use of a compound that inhibits HIF prolyl hydroxylase activity in manufacturing a medicament for increasing dopaminergic neurogenesis.

Increased neurogenesis, particularly increased dopaminergic neurogenesis, may also be beneficial in treatment and functional recovery in a subject having progressive neurological disorders including various neurodegenerative disorders, such as Parkinson's disease and essential tremor. In one embodiment, the present invention provides methods for increasing dopaminergic neurogenesis in a subject having a neurodegenerative disease, the method comprising administering to the subject a compound that inhibits

HIF prolyl hydroxylase activity. In one embodiment, the present invention provides methods for increasing dopaminergic neurogenesis in a subject having Parkinson's disease, the method comprising administering to the subject a compound that inhibits HEF prolyl hydroxylase activity. In one embodiment, the present invention provides methods for increasing dopaminergic neurogenesis in a subject having essential tremor, the method comprising administering to the subject a compound that inhibits HEF prolyl hydroxylase activity. In another embodiment, the invention provides for use of a compound that inhibits HEF prolyl hydroxylase activity in manufacturing a medicament for treatment of neurodegnerative disease such as Parkinson's disease, essential tremor, and various types of kinetic tremors. The present methods and compounds are also useful for treating various conditions wherein increased neurogenesis would be beneficial, such as chronic stress disorders, e.g., post-traumatic stress disorder (PTSD); depression, Cushing's disease, addiction, epilepsy, and schizophrenia. The present methods and compounds are also useful for treating mild cognitive impairment (MCI) or dementia, e.g., age-related dementia. In one embodiment, the present invention provides methods for increasing neurogenesis in a subject having cognitive impairment, e.g., mild cognitive impairment, the method comprising administering to the subject a compound that inhibits HIF prolyl hydroxylase activity. In another embodiment, the invention provides for use of a compound that inhibits HIF prolyl hydroxylase activity in manufacturing a medicament for treatment of mild cognitive impairment.

Compounds and methods of the present invention are effective at increasing neurogenesis, as demonstrated, in part, by increased numbers of proliferating neural cells following administration of compound of the present invention. Proliferating neural cells can be identified by their expression of neuron-specific class III β-tubulin (TUJl) and doublecortin (DCX). TUJl is expressed in neurons of the peripheral and central nervous systems and contributes to microtubule stability in neuronal cell bodies and axons. DCX is a microtubule-associated protein expressed almost exclusively in immature (e.g., developing) neurons, and thus is a marker for neurogenesis. Neural precursors begin to express DCX shortly after exiting the cell cycle, and continue to express DCX as the cells mature into neurons. (Couillard-Despres et al. (2005) Eur J Neurosci 21:1-14.)

In one embodiment, the present invention provides a method for increasing expression of TUJl in a neural cell, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing expression of TUJl in the neural cell. In another embodiment, the present invention provides a method for increasing expression of DCX in a neural cell, the method comprising administering to or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing expression of DCX in the neural cell.

Compounds and methods of the present invention are effective at increasing neurogenesis, as demonstrated, in part, by increased expression of various neural cell markers following administration of compound of the present invention. Neural cell markers associated with neurogenesis include increased expression of neuron-specific enolase (NSE) or increased expression of microtubule-associated protein 2 (MAP2). MAP2 is a microtubule associated protein found exclusively in nerve cells. In one embodiment, the present invention provides a method for increasing expression of MAP2 in a neural cell, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing expression of MAP2 in the neural cell. In another embodiment, the present invention also provides a method for increasing expression of neuron specific enolase (NSE) in a neural cell, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing expression of neuron specific enolase in the neural cell.

The present invention further provides a method for increasing neural cell differentiation, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing neural cell differentiation. In particular embodiments, the present invention provides methods for increasing neural cell differentiation in a subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HIF prolyl hydroxylase activity. The present invention also provides compounds for use in manufacturing a medicament for increasing neural cell differentiation, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme. In certain embodiments, the neural cell in these methods is a neural progenitor cell such as, for example, a mesencephalic neural progenitor cell or a cortical (i.e., frontal) neural progenitor cell. In other embodiments, the neural cell in these methods is a neural stem cell, a neural progenitor cell, or a neural precursor cell.

hi another embodiment, the present invention provides a method for increasing dopaminergic differentiation of neural cells, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing dopaminergic differentiation of the neural cell. In some embodiments, the present invention provides methods for increasing dopaminergic differentiation of neural cells in a subject in need, the method comprising administering to the subject an effective amount of a compound that inhibits HIF prolyl hydroxylase activity. In yet another embodiment, the present invention provides a method for increasing dopaminergic neurons (i.e., increasing the number of dopaminergic neurons), the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing dopaminergic neurons.

Dopaminergic neurons are neurons whose primary neurotransmitter is dopamine. Dopaminergic neurons are present chiefly in the ventral tegmental area of the midbrain, substantia nigra pars compacta, and arcuate nucleus of the hypothalamus. Dopaminergic neurons are associated with numerous functions in the brain, including important roles in behavior and cognition, motor activity, motivation and reward, inhibition of prolactin production (involved in lactation), sleep, mood, attention, and learning.

Dopaminergic differentiation and identification of dopaminergic neurons can be determined by any measure known to those skilled in the art. For example, dopaminergic differentiation and dopaminergic neurons are identified by the presence of characteristic and identifying dopaminergic markers, such as, for example, tyrosine hydroxylase (TH) and microtubule-associated protein 2 (MAP2). Tyrosine hydroxylase is the enzyme responsible for catalyzing the conversion of the L-tyrosine to dihydrophenylalanine, a precursor for dopamine. (Masserano and Weiner (1983) MoI Cell Biochem 53- 54:129-152). MAP2 is a microtubule associated protein found exclusively in nerve cells.

The present invention also provides a method for increasing expression of tyrosine hydroxylase in a neural cell, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing expression of tyrosine hydroxylase in the neural cell.

In another embodiment, the present invention provides a method for increasing neural cell proliferation, the method comprising administering to a subject or contacting a neural cell with an effective amount of a compound that inhibits the activity of a HIF prolyl hydroxylase enzyme, thereby increasing neural cell proliferation. The present invention also provides compounds for use in manufacturing a medicament for increasing neural cell proliferation, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme. In certain embodiments, the neural cell in these methods is a neural progenitor cell such as, for example, a mesencephalic neural progenitor cell or a cortical (i.e., frontal) neural progenitor cell. In yet other embodiments, the neural cell in these methods is a neural stem cell, a neural progenitor cell, or a neural precursor cell.

Neural cell proliferation can be identified by any measure known to one skilled in the art. For example, proliferating neural cells are identified by the presence of the neural cell marker neuron-specific class IQ β-tubulin (TUJl) and the cell proliferating marker Ki67. TUJl is expressed in neurons of the peripheral and central nervous systems and contributes to microtubule stability in neuronal cell bodies and axons. Ki67 is a cellular marker for proliferation, is present in all active phases of the cell cycle (Gl, S, G2, and mitosis), but is absent in resting cells (GO). Ki67 is used as a marker to determine the growth fraction of a given cell population. (See, e.g., Gerdes et al. (1983) Int J Cancer 31 : 13-20 and Scholzen and Gerdes (2000) J Cell Physiol 182:311-322.)

Methods and compounds of the present invention were also effective at increasing neurites in neural progenitor cells and effective at increasing VEGF expression in neural progenitor cells. Subjects

The present invention relates to methods for increasing neurogenesis. The invention is applicable to a variety of different organisms, including, for example, vertebrates, large animals, and primates. In a preferred embodiment, the subject is a mammalian subject, and in a most preferred embodiment, the subject is a human subject. However, although medical applications with humans are clearly foreseen, veterinary applications are also envisaged herein.

In certain embodiments, the present methods of treatment involve administration of an effective amount of a compound to a subject in need, wherein the compound inhibits the activity of a HIF prolyl hydroxylase enzyme, and wherein the subject would benefit from increased neurogenesis, neural cell differentiation, dopaminergic differentiation, or neural cell proliferation. In some embodiments, the subject has a neurodegenerative disease wherein increased neurogenesis would be beneficial. In particular embodiments, the subject has Parkinson's disease, essential tremor (ET), or any type of kinetic tremor. In an additional embodiment, the subject is a subject with an altered dopaminergic response. In further emobidments, the subject has a chronic neurological disorder such as epilepsy. In other embodiments, the subject has a phycological or neurological impairment, such as chronic stress disorders, e.g., post-traumatic stress disorder (PTSD); depression, Cushing's disease, addiction, epilepsy, or schizophrenia. In particular embodiments, the subject has mild cognitive impairment (MCI) or dementia, e.g., age-related dementia.

Compounds

Compounds for use in the methods or medicaments provided herein stabilize hypoxia-inducible factor alpha (HIFα) and are inhibitors of hypoxia-inducible factor (HIF) prolyl hydroxylase enzymes. A compound that inhibits the activity of HIF prolyl hydroxylase enzyme refers to any compound that reduces or otherwise moldulates the activity of at least one HIF prolyl hydroxylase enzyme. The term

"HIF prolyl hydroxylase," as used herein, refers to any enzyme that is capable of hydroxylating a proline residue within an alpha subunit of HIF. Such HIF prolyl hydroxylases include protein members of the EGL-9 (EGLN) 2-oxoglutarate- and iron-dependent dioxygenase family described by Taylor (2001) Gene 275: 125-132; and characterized by Aravind and Koonin (2001) Genome Biol 2:RESEARCH0007; Epstein et al. (2001) Cell 107:43-54; and Bruick and McKnight (2001) Science 294: 1337-1340.

Functionally, prolyl hydroxylase inhibitors for use in the methods of the present invention are defined by their ability to inhibit an activity of a 2-oxoglutarate dioxygenase enzyme, wherein the enzyme has specific activity toward hypoxia inducible factor. Such compounds are defined herein as prolyl hydroxylase inhibitors (PHIs). Preferably, the PHIs for use in the invention are small molecule compounds. A compound that inhibits the activity of a HIF prolyl hydroxylase enzyme refers to any compound that reduces or otherwise modulates the activity of at least one HIF prolyl hydroxylase enzyme. A compound may additionally show inhibitory activity toward one or more other 2- oxoglutarate- and iron-dependent dioxygenase enzymes, e.g. factor inhibiting HIF (FIH; GenBank Accession No. AAL27308), procollagen prolyl 4-hydroxylase (CP4H), etc.

It is contemplated herein that a "compound that inhibits HIF prolyl hydroxylase" suitable for use in the claimed methods can be any compound that inhibits HIF prolyl hydroxylase activity. As noted herein, the compound that inhibits HIF prolyl hydroxylase activity can be a structural mimetic of 2-oxoglutarate. In some embodiments, the compound is a structural mimetic of 2-oxoglutarate that inhibits HIF prolyl hydroxylase activity competitively with respect to 2-oxoglutarate. In particular embodiments, compounds used in the present methods and medicaments provided herein are structural mimetics of 2-oxoglutarate, wherein the compound inhibits the target HIF prolyl hydroxylase enzyme competitively with respect to 2- oxoglutarate and noncompetitively with respect to iron.

In some structural embodiments, a compound suitable for use in the present invention is a cyclic carboxamide. The cyclic group is variously a carbocycle or a heterocycle. It is specifically contemplated that the cyclic group may contain additional substitutions at ring positions not occupied by the carboxamide moiety; for example, substitution of one or more atoms within the ring with a hydroxyl (- OH) or oxo (=0) group.

Accordingly, in certain embodiments, the compound is a carbocyclic carboxamide. The carbocyclic group can be a single ring group, e.g., a benzene, or can contain multiple condensed rings, e.g., a napthalene.

In particular embodiments, a compound suitable for use in the present invention is a heterocyclic carboxamide. In selected embodiments, the heterocycle can be a single ring, for example, a pyridine, a pyrimidine, or a pyridazine. In other embodiments, the specified heterocyclic structure is a multiple condensed ring, for example, a quinoline, a cinnoline, an isoquinoline, a pyrrolopyridine, a napthyridine, a β-carboline, a chromene (coumarin), or a thiochromene (thiocoumarin).

Carboxamide compounds particularly suitable for use in the present invention include carboxamides substituted at the amide to form a carbonyl glycine. Therefore, in certain embodiments, a compound for use in the present invention is a cyclic carbonyl glycine, and in particular, a carbocyclic carbonyl glycine or a heterocyclic carbonyl glycine. Specifically encompassed by the term "carbonyl glycine" are structural and functional analogs thereof, including, in particular, carbonyl glycineamides (wherein the carboxyl moiety on the glycine is replaced with carboxamide). Also encompassed are prodrugs thereof, such as carbonyl glycine esters (wherein the carboxyl moiety is esterified with a substituent such as an alkyl, e.g., methyl). In certain embodiments of the present invention, specific substitution at the α carbon of the glycine of a suitable heterocyclic carbonyl glycine compound results in replacement of the glycine with a comparable amino acid selected from the group consisting of alanine, valine, leucine, and isoleucine.

In certain embodiments of the present invention, a cyclic carboxamide for use in the present invention is a heterocyclic carboxamide and, more specifically, an isoquinoline carboxamide. The carboxamide can be positioned on the isoquinoline at any stereochemically appropriate point on the heterocycle. Isoquinoline carboxamides particularly suited for use in the present invention include isoquinoline-3-carboxamides. Examples of such isoquinoline carboxamides include [(4-hydroxy-7-phenylsulfanyl-isoquinoline-3- carbonyl)-amino] -acetic acid (Compound A); [(4-hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]- acetic acid (Compound B); {[l-cyano-6-(2,6-dimethyl-phenoxy)-4-hydroxy-isoquinoline-3- carbonyl]- amino} -acetic acid (Compound C), and other compounds easily identified by those skilled in the art, including those described and claimed in, e.g., United States Patent No. 6,093,730 and International Publication Nos. WO 2004/108681 and WO 2007/090068.

In some embodiments, an isoquinoline carboxamide according to the present invention is additionally substituted with a hydroxyl group; specifically a hydroxy isoquinoline carboxamide. The hydroxyl can be positioned at any stereochemically appropriate point on the heterocycle. Accordingly, isoquinoline carboxamides suitable for use in the present invention include 4-hydroxy-isoquinoline-3 -carboxamides. Examples of such compounds include Compound A, Compound B, Compound C, and other compounds easily identified by those skilled in the art, including those described and claimed in, e.g., United States Patent No. 6,093,730 and International Publication Nos. WO 2004/108681 and WO 2007/090068.

In certain embodiments in which a compound of the invention is an isoquinoline carboxamide, the amide on the carboxamide moiety of the isoquinoline carboxamide is substituted to form a glycine, and the compound for use in the present invention is an isoquinoline carbonyl glycine. As the present invention particularly encompasses use of isoquinoline-3-carboxamides, isoquinoline-3 -carbonyl glycines are specifically contemplated herein, as are more substituted examples thereof, including 4-hydroxy- isoquinoline-3-carbonyl glycines, etc. Examples of such compounds include Compound A, Compound B, Compound C, and other compounds easily identified by those skilled in the art, including those described and claimed in, e.g., United States Patent No. 6,093,730 and in International Publication Nos. WO 2004/108681 and WO 2007/090068.

In other embodiments of the present invention, a cyclic carboxamide according to the present invention is a heterocyclic carboxamide and, more specifically, a pyrrolopyridazine carboxamide. The carboxamide can be positioned at any stereochemically appropriate point on the heterocycle. Pyrrolopyridazine carboxamides suitable for use in the present invention include pyrrolopyridazine-3-carboxamides. Examples of such pyrrolopyridazine carboxamides include {[4-hydroxy-2-oxo-l-(4-trifluoromethyl- benzyO-l^-dihydro-pyrrolofl^-blpyridazine-S-carbonylJ-aminoJ -acetic acid (Compound D) and other compounds readily identified by those skilled in the art, including those described and claimed in, e.g., International Application No. PCT/US09/54473.

In some embodiments, a pyrrolopyridazine carboxamide according to the present invention is additionally substituted with an oxo group; specifically, an oxo-pyrrolopyridazine carboxamide (pyrrolopyridazinone carboxamide). The oxo can be positioned at any stereochemically appropriate point on the heterocycle. Pyrrolopyridazine carboxamides particularly suited for use in the present invention include 2-oxo- pyrrolopyridazine-3 -carboxamides. Examples of such compounds include Compound D and other compounds easily identified by those skilled in the art, including those described and claimed in, e.g., International Application No. PCT/US09/54473.

In some embodiments, the pyrrolopyridazine carboxamide according to the present invention is additionally substituted with a hydroxyl group; specifically, a hydroxy pyrrolopyridazine carboxamide.

The hydroxyl can be positioned at any stereochemically appropriate point on the heterocycle. Pyrrolopyridazine carboxamides suitable for use in the present invention include 4-hydroxy- pyrrolopyridazine-3-carboxamides. Examples of such compounds include Compound D and other compounds easily identified by those skilled in the art, including those described and claimed in, e.g., International Application No. PCT/US09/54473.

In particular embodiments, a pyrrolopyridazine carboxamide according to the present invention is additionally substituted with a hydroxyl group and an oxo group; specifically, specifically, a hydroxy oxo-pyrrolopyridazine carboxamide (hydroxy pyrrolopyridazinone carboxamide). The oxo and hydroxyl can be independently positioned at any stereochemically appropriate point on the heterocycle.

Pyrrolopyridazine carboxamides suitable for use in the present invention include 4-hydroxy-2-oxo- pyrrolopyridazine-3 -carboxamide. Examples of such compounds include Compound D and other compounds easily identified by those skilled in the art, including those described and claimed in, e.g., International Application No. PCT/US09/54473.

In certain embodiments in which a compound of the invention is a pyrrolopyridazine carboxamide, the amide on the carboxamide moiety of the pyrrolopyridazine carboxamide is substituted to form a glycine, and the compound for use in the present invention is a pyrrolopyridazine carbonyl glycine. As the present invention particularly encompasses use of pyrrolopyridazine-3 -carboxamides, pyrrolopyridazine -3- carbonyl glycines are specifically contemplated herein, as are more substituted examples thereof, including 4-hydroxy-pyrrolopyridazine-3-carbonyl glycines, 2-oxo-pyrrolopyridazine-3-carbonyl glycines, and 4-hydroxy-2-oxo-pyrrolopyridazine-3 -carboxamides. Examples of such include Compound D and other compounds easily identified by those skilled in the art, including those described and claimed in, e.g., International Application No. PCT/US09/54473.

Additional examples of compounds suitable for use according to the present invention are presented below.

In other particular embodiments, a compound of the invention is a heterocyclic carbonyl glycine of formula VI.

wherein R is an optionally substituted heterocyclic moiety.

In specific embodiments, such compounds include, but are not limited to, substituted 3-hydroxy-pyridine- 2-carbonyl-glycines, 4-hydroxy-pyridazine-3-carbonyl-glycines, 3-hydroxy-quinoline-2-carbonyl- glycines, 4-hydroxy-2-oxo-l,2-dihydro-quinoline-3 -carbonyl -glycines, 4-hydroxy-2-oxo-l,2-dihydro- naphthyridine-3-carbonyl-glycines, 8-hydroxy-6-oxo-4,6-dihydro-pyridopyrazine-7-carbonyl-glycin es, 4- hydroxy-isoquinoline-3 -carbonyl -glycines, 4-hydroxy-cinnoline-3 -carbonyl-glycines, 7-hydroxy- thienopyridine-6-carbonyl-glycines, 4-hydroxy-thienopyridine-5-carbonyl-glycines, 7-hydroxy- thiazolopyridine-6-carbonyl-glycines, 4-hydroxy-thiazolopyridine-5 -carbonyl-glycines, 7-hydroxy- pyrrolopyridine-6-carbonyl-glycines, and 4-hydroxy-pyrrolopyridine-5-carbonyl-glycines. Compounds can be identified for use in the present embodiments by measuring inhibitory activity of the compound on a HIF prolyl hydroxylase enzyme, e.g., using an enzyme assay as described herein. More generally, compounds can be identified for use in the present embodiments by measuring HIF stabilization induced by the compound, e.g., using a cell-based assay as described herein.

In certain embodiments, the heterocyclic carbonyl glycine is a quinoline carboxamide, an isoquinoline carboxamide, a pyridine carboxamide, a cinnoline carboxamide, or a beta-carboline carboxamide. Heterocyclic carbonyl glycines effectively stabilize HIFα. In preferred aspects, a compound of the invention is a compound that inhibits prolyl hydroxylase activity (e.g., a prolyl hydroxylase inhibitor). In more preferred aspects, a compound of the invention is a compound that inhibits HIF prolyl hydroxylase activity. Prolyl hydroxylase inhibitors (PHIs) specifically contemplated for use in the present methods are described, e.g., in Majamaa et al., supra; Kivirikko and Myllyharju (1998) Matrix Biol 16:357-368; Bickel et al. (1998) Hepatology 28:404-411; Friedman et al. (2000) Proc Natl Acad Sci USA 97:4736- 4741; Franklin (1991) Biochem Soc Trans 19):812 815; Franklin et al. (2001) Biochem J 353:333-338. Examples of compounds that may be used in the methods and medicaments provided herein can be found, e.g., in Majamaa et al. (1984) Eur. J. Biochem. 138:239-245; Majamaa et al. (1985) Biochem. J. 229:127- 133; Kivirikko, and Myllyharju (1998) Matrix Biol. 16:357-368; Bickel et al. (1998) Hepatology 28:404- 411; Friedman et al. (2000) Proc. Natl. Acad. Sci. USA 91:4136 Al 41; Franklin (1991) Biochem. Soc. Trans. 19):812-815; and Franklin et al. (2001) Biochem. J. 353:333-338. Additionally, compounds that inhibit HIF prolyl hydroxylase enzyme activity or that stabilize HIFαhave been described in, e.g.,

International Publication Nos. WO 2003/049686, WO 2002/074981, WO 03/053977, WO 2003/080566, WO 2004/108121, WO 2004/108681, WO 2006/094292, WO 2007/038571, WO 2007/090068, WO 2007/070359, WO 2007/103905, and WO 2007/115315. All compounds listed in the above-patent and patent applications are hereby incorporated into the present application by reference herein in their entirety.

In various embodiments, a compound for use in the present methods is a heterocyclic carbonyl glycine, in particular, a heterocyclic carbonyl glycine of Formula VI. In certain embodiments, the compound used in the present methods is a compound selected from the group consisting of the compounds of Formula I, Formula π, Formula HI, Formula IV, Formula V, and Formula VI. Formula I includes, but is not limited to, compounds of

Formulae Ia, Ib, Ic, and Id. Formula HI includes, but is not limited to, the compounds of Formula HIa. Formula IV includes, but is not limited to, compounds of Formulae IVA, IVB, IVC, and IVD. Formula V includes, but is not limited to, compounds of Formulae VA, VB, VC, and VD.

As stated, supra, compounds suitable for use in the present invention include isoquinoline carboxamides.

In various emobidments, isoquinoline carboxamides according to the present invention are isoquinoline- 3-carboxamides. In one embodiment, a compound for use in the methods of the present invention is [(4- Hydroxy-7-phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-ace tic acid (Compound A), [(4-Hydroxy-7-phenoxy-isoquinoline-3-carbonyl)-amino]-acetic acid (Compound B), {[l-Cyano-6-(2,6- dimethyl-phenoxy)-4-hydroxy-isoquinoline-3-carbonyl]-amino} -acetic acid (Compound C), or {[4-

Hydroxy-2 -oxo-1 -(4-trifluoromethyl-benzyl)-l,2-dihydro-pyrrolo[l,2-b]pyrida zine-3-carbonyl]-amino}- acetic acid (Compound D).

In certain embodiments, compounds used in the methods of the invention are heterocyclic carboxamides selected from a compound of the formula (I)

wherein A is 1,2-arylidene, 1,3-arylidene, 1,4-arylidene; or (d-C ₄ )-alkylene, optionally substituted by one or two halogen, cyano, nitro, trifluoromethyl, (d-C ₆ )-alkyl, (Ci-C ₆ )-hydroxyalkyl, (C ₁ - C ₆ )-alkoxy, -O-[CH ₂ ]χ-C _f H _(2f+ i. _g >Hal _g , (Ci-C ₆ )-fluoroalkoxy, (C ₁ -C ₈ )-fluoroalkenyloxy, (C _r C ₈ )-fluoroalkynyloxy, -OCF ₂ Cl, -0-CF ₂ -CHFCl; (Ci-C ₆ )-alkylmercapto, (C ₁ -C ₆ )- alkylsulfinyl, (C ₁ -C ₆ )-alkylsulfonyl, (C ₁ -C ₆ )-alkylcarbonyl, (d-C ₆ )-alkoxycarbonyl, carbamoyl, N-(C ₁ -C ₄ )-alkylcarbamoyl, N,N-di-(C ₁ -C ₄ )-alkylcarbamoyl, (C ₁ -C ₆ )- alkylcarbonyloxy, (C ₃ -C8)-cycloalkyl, phenyl, benzyl, phenoxy, benzyloxy, anilino, N- methylanilino, phenylmercapto, phenylsulfonyl, phenylsulfinyl, sulfamoyl, N-(Ci-C ₄ )- alkylsulfamoyl, N,N-di-(C ₁ -C ₄ )-alkylsulfamoyl; or by a substituted (C ₆ -C ₁₂ )-aryloxy, (C ₇ - C _π )-aralkyloxy, (C ₆ -C ₁₂ )-aryl, (C ₇ -C ₁ 0-aralkyl radical, which carries in the aryl moiety one to five identical or different substituents selected from halogen, cyano, nitro, trifluoromethyl, (C,-C ₆ )-alkyl, (C _r C ₆ )-alkoxy, -O-[CH ₂ ] _x -C _f H _(2f+1 . _g) Hal _g , -OCF ₂ Cl, -O- CF ₂ -CHFCl, (d-C^-alkylmercapto, (d-C ₆ )-alkylsulfmyl, (C,-C ₆ )-alkylsulfonyl, (C ₁ - C ₆ )-alkylcarbonyl, (Ci-C ₆ )-alkoxycarbonyl, carbamoyl, N-(C ₁ -C ₄ )-alkylcarbamoyl, N,N- di-(C ₁ -C ₄ )-alkylcarbamoyl, (C _! -C ₆ )-alkylcarbonyloxy, (C ₃ -C ₈ )-cycloalkyl, sulfamoyl, N-

(C _r C ₄ )-alkylsulfamoyl, N,N-di-(C ₁ -C ₄ )-alkylsulfamoyl; or wherein A is -CR ⁵ R ⁶ and R ⁵ and R ⁶ are each independently selected from hydrogen, (C ₁ -C ₆ )-alkyl, (C ₃ -C ₇ )-cycloalkyl, aryl, or a substituent of the α-carbon atom of an α-amino acid, wherein the amino acid is a natural L-amino acid or its D-isomer. B is -CO ₂ H, -NH ₂ , -NHSO ₂ CF ₃ , tetrazolyl, imidazolyl, 3-hydroxyisoxazolyl, -CONHCOR'",

-CONHSOR'", or -CONHSO ₂ R ¹ ", where R'" is aryl, heteroaryl, (C ₃ -C ₇ )-cycloalkyl, or (d-C ₄ )-alkyl, optionally monosubstituted by (C ₆ -C ₁₂ )-aryl, heteroaryl, OH, SH, (C ₁ -C ₄ )- alkyl, (C,-C ₄ )-alkoxy, (C,-C ₄ )-thioalkyl, (C ₁ -C ₄ )-sulfinyl, (C _r C ₄ )-sulfonyl, CF ₃ , Cl, Br, F, I, N02, -COOH, (C ₂ -C ₅ )-alkoxycarbonyl, NH ₂ , mono-(C ₁ -C ₄ -alkyl)-amino, di-(C _r C ₄ - alkyl)-amino, or (C _r C ₄ )-perfluoroalkyl; or wherein B is a CO ₂ -G carboxyl radical, where

G is a radical of an alcohol G-OH in which G is selected from (d-C ₂₀ )-alkyl radical, (C ₃ - C ₈ ) cycloalkyl radical, (C ₂ -C ₂₀ )-alkenyl radical, (C ₃ -C ₈ )-cycloalkenyl radical, retinyl radical, (C ₂ -C ₂₀ )-alkynyl radical, (C ₄ -C ₂₀ )-alkenynyl radical, where the alkenyl, cycloalkenyl, alkynyl, and alkenynyl radicals contain one or more multiple bonds; (C ₆ - C ₁₆ )-carbocyclic aryl radical, (C ₇ -C ₁₆ )-carbocyclic aralkyl radical, heteroaryl radical, or heteroaralkyl radical, wherein a heteroaryl radical or heteroaryl moiety of a heteroaralkyl radical contains 5 or 6 ring atoms; and wherein radicals defined for G are substituted by one or more hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl, (C ₁ -C] ₂ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C ₅ -C ₈ )-cycloalkenyl, (C ₆ -C ₁₂ )-aryl, (C ₇ -C ₁₆ )-aralkyl, (C ₂ -C ₁₂ )- alkenyl, (C ₂ -C _I2 )-alkynyl, (C,-C ₁₂ )-alkoxy, (d-C^-alkoxy-td-Q^-alkyl, (C ₁ -C ₁₂ )- alkoxy-(d-C ₎₂ )-alkoxy, (C ₆ -C ₁₂ )-aryloxy, (C ₇ -C ₁₆ )-aralkyloxy, (d-C ₈ )-hydroxyalkyl, -O- [CH ₂ ] _x -C _f H _(2f+1-g) -F _g , -OCF ₂ Cl, -OCF ₂ -CHFCl, (C,-C ₁₂ )-alkylcarbonyl, (C ₃ -C ₈ )- cycloalkylcarbonyl, (C ₆ -Ci ₂ )-arylcarbonyl, (C ₇ -Ci ₆ )-aralkylcarbonyl, cinnamoyl, (C ₂ - Ci ₂ )-alkenylcarbonyl, (C ₂ -Ci ₂ )-alkynylcarbonyl, (Ci-Ci ₂ )-alkoxycarbonyl, (C _r Ci ₂ )- alkoxy-(Ci-Ci ₂ )-alkoxycarbonyl, (C ₆ -Ci ₂ )-aryloxycarbonyl, (C ₇ -Ci ₆ )-aralkoxycarbonyl, (C ₃ -C ₈ )-cycloalkoxycarbonyl, (C ₂ -C ₁₂ )-alkenyloxycarbonyl, (C ₂ -Ci ₂ )- alkynyloxycarbonyl, acyloxy, (Ci-Ci ₂ )-alkoxycarbonyloxy, (Ci-Ci ₂ )-alkoxy-(C _r Ci ₂ )- alkoxycarbonyloxy, (C ₆ -C ₎₂ )-aryloxycarbonyloxy, (C ₇ -Ci ₆ ) aralkyloxycarbonyloxy, (C ₃ - C ₈ )-cycloalkoxycarbonyloxy, (C ₂ -Ci ₂ )-alkenyloxycarbonyloxy, (C ₂ -Ci ₂ )- alkynyloxycarbonyloxy, carbamoyl, N-(Ci-Ci ₂ )-alkylcarbamoyl, N.N-di(Ci-Ci ₂ )- alkylcarbamoyl, N-(C ₃ -C ₈ )-cycloalkyl-carbamoyl, N-(C ₆ -Ci ₆ )-arylcarbamoyl, N-(C ₇ -Ci ₆ )- aralkylcarbamoyl, N-(Ci-Ci ₀ )-alkyl-N-(C ₆ -Ci ₆ )-arylcarbamoyl, N-(C,-Ci _O )-alkyl-N-(C ₇ -

C ₁₆ )-aralkylcarbamoyl, N-((C,-Cio)-alkoxy-(Ci-Cio)-alkyl)-carbamoyl, N-((C ₆ -Ci ₂ )- aryloxy-(C]-Cio)alkyl)-carbamoyl, N-((C ₇ -Ci ₆ )-aralkyloxy-(Ci-Cio)-alkyl)-carbamoyl, N- (C ₁ -C ₁₀ )-alkyl-N-((C ₁ -Cio)-alkoxy-(C,-Cio)-alkyl)-carbamoyl, N-(C,-Cio)-alkyl-N-((C ₆ - C _I6 )-aryloxy-(Ci-Cio)-alkyl)-carbamoyl, N-(Ci-Cio)-alkyl-N-((C ₇ -C, ₆ )-aralkyloxy-(Ci- Cio)-alkyl)-carbamoyl, carbamoyloxy, N-(Ci-Ci ₂ )-alkylcarbamoyloxy, N.N-di-(C _r Ci ₂ )- alkylcarbamoyloxy, N-(C ₃ -C ₈ )-cycloalkylcarbamoyloxy, N-(C ₆ -Ci ₂ )-arylcarbamoyloxy, N-(C ₇ -C ₁₆ )-aralkylcarbamoyloxy, N-(Ci-Cio)-alkyl-N-(C ₆ -C ₁₂ )-arylcarbamoyloxy, N(Cr Cio)-alkyl-N-(C ₇ -Ci ₆ )-aralkylcarbamoyloxy, N-((Ci-Ci ₀ )-alkyl)-carbamoyloxy, N-((C ₆ - Ci ₂ )-aryloxy-(Ci-Cio)-alkyl)-carbamoyloxy, N-((C ₇ -Ci ₆ )-aralkyloxy-(C _r Ci ₀ )-alkyl)- carbamoyloxy, N-(Ci-Cio)-alkyl-N-((Ci-Ci ₀ )-alkoxy-(Ci-C _IO )-alkyl)-carbamoyloxy, N-

(Ci-C,o)-alkyl-N-((C ₆ -Ci ₂ )-aryloxy-(C,-C ₁₀ )-alkyl)-carbamoyloxy, N-(Ci-C ₁₀ )-alkyl-N- ((C ₇ -Ci ₆ )-aralkyloxy-(Ci-Cio)-alkyl)-carbamoyloxy, amino, (Ci-C ₎₂ )-alkylamino, di-(C _r Ci ₂ )-alkylamino, (C ₃ -C ₈ )-cycloalkylamino, (C ₂ -Ci ₂ )-alkenylamino, (C ₂ -Ci ₂ )- alkynylamino, N-(C ₆ -C i ₂ )-arylamino, N-(C-Ci i)-aralkylamino, N-alkyl-aralkylamino, N- alkyl-arylamino, (Ci-Ci ₂ )-alkoxyamino, (C]-Ci ₂ )-alkoxy-N-(C ₁ -Ci ₀ )-alkylamino, (Ci-

Ci ₂ )-alkylcarbonylamino, (C ₃ -C ₈ )-cycloalkylcarbonylamino, (C ₆ -Ci ₂ ) arylcarbonylamino, (C ₇ -Ci ₆ )-aralkylcarbonylamino, (C _r Ci ₂ )-alkylcarbonyl-N-(Ci-Cio)-alkylamino, (C ₃ -C ₈ )- cycloalkylcarbonyl-N-(Ci-Cio)-alkylamino, (C ₆ -Ci ₂ )-arylcarbonyl-N-(Ci-Ci ₀ )alkylamino, (C ₇ -Cii)-aralkylcarbonyl-N-(Ci-Cio)-alkylamino, (Ci-Ci ₂ )-alkylcarbonylamino-(Ci-C ₈ )- alkyl, (C ₃ -C ₈ )-cycloalkylcarbonylamino-(Ci-C ₈ )alkyl, (C ₆ -Ci ₂ )-arylcarbonylamino-(C _r

C ₈ )-alkyl, (C ₇ -Ci ₂ )-aralkylcarbonylamino(Ci-C ₈ )-alkyl, amino-(Ci-Ci ₀ )-alkyl, N-(Ci-Ci ₀ ) alkylamino-(C,-Cio)-alkyl, N.N-di-(C,-C, ₀ )-alkylamino-(Ci-Cio)-alkyl, (C ₃ - C ₈ )cycloalkylamino-(Ci-C,o)-alkyl, (Ci-C, ₂ )-alkylmercapto, (Ci-Ci ₂ )-alkylsulfinyl, (C ₁ - Ci ₂ )-alkylsulfonyl, (C ₆ -Ci ₆ )-arylmercapto, (C ₆ -Ci ₆ )-arylsulfinyl, (C ₆ -Ci ₂ )-arylsulfonyl, (C ₇ -Ci ₆ )-aralkylmercapto, (C ₇ -Ci ₆ )-aralkylsulfinyl, (C ₇ -Ci ₆ )-aralkylsulfonyl, sulfamoyl,

N-(Ci-C,o)-alkylsulfamoyl, N.N-di(C _r Cio)-alkylsulfamoyl, (C ₃ -C ₈ )-cycloalkylsulfamoyl, N-(C ₆ -C ₁₂ )-alkylsulfamoyl, N-(C ₇ -C, ₆ )-aralkylsulfamoyl, N-(C _I -Cio)-alkyl-N-(C ₆ -C, ₂ )- arylsulfamoyl, N-(Ci-Cio)-alkyl-N-(C ₇ -Ci ₆ )-aralkylsulfamoyl, (C,-Ci ₀ )-alkylsulfonamido, N-((C ₁ -C,o)-alkyl)-(C,-Cio)-alkylsulfonamido, (C ₇ -Ci ₆ )-aralkylsulfonamido, or N-((C,- Cio)-alkyl-(C ₇ -Ci ₆ )-aralkylsulfonamido; wherein radicals which are aryl or contain an aryl moiety, may be substituted on the aryl by one to five identical or different hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl, (Ci-C _]2 )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C ₆ -

C ₁₂ )-aryl, (C ₇ -C ₁₆ )-aralkyl, (C,-C ₁₂ )-alkoxy, (C ₁ -C ₁₂ )-alkoxy-(C ₁ -C ₁₂ )alkyl, (C ₁ -C ₁₂ )- alkoxy-(C] Ci ₂ )alkoxy, (C ₆ -C ₁₂ )-aryloxy, (C ₇ -C, ₆ )-aralkyloxy, (Ci-Cg)-hydroxyalkyl, (C ₁ - Ci ₂ )-alkylcarbonyl, (C ₃ -C ₈ )-cycloalkyl-carbonyl, (C ₆ -C ₁₂ )-arylcarbonyl, (C ₇ -C ₁₆ ) aralkylcarbonyl, (d-C ₁₂ )-alkoxycarbonyl, (C ₁ -C ₁₂ )-alkoxy-(C ₁ -C ₁₂ )-alkoxycarbonyl, (C ₆ - C] ₂ )-aryloxycarbonyl, (C ₇ -C ₁₆ )-aralkoxycarbonyl, (C ₃ -C ₈ )-cycloalkoxycarbonyl, (C ₂ -C ₁₂ )- alkenyloxycarbonyl, (C ₂ -C ₁₂ )-alkynyloxycarbonyl, (C _r C ₁₂ )-alkylcarbonyloxy, (C ₃ -C ₈ )- cycloalkylcarbonyloxy, (C ₆ -C ₁₂ )-arylcarbonyloxy, (C ₇ -C ₁₆ )-aralkylcarbonyloxy, cinnamoyloxy, (C ₂ -C ₁₂ )-alkenylcarbonyloxy, (C ₂ -C ₁₂ )-alkynylcarbonyloxy, (C ₁ -C ₁₂ )- alkoxycarbonyloxy, (C ] -Ci ₂ )-alkoxy-(C i -C ₁₂ )-alkoxycarbonyloxy, (C ₆ -C _{t 2} )- aryloxycarbonyloxy, (C ₇ -C i ₆ )-aralkyloxycarbonyloxy, (C ₃ -C ₈ )-cycloalkoxycarbonyloxy,

(C ₂ -Ci ₂ )-alkenyloxycarbonyloxy, (C ₂ -C ₁₂ )-alkynyloxycarbonyloxy, carbamoyl, N-(C ₁ - Ci ₂ )-alkylcarbamoyl, N.N-di-(Ci-C ₁₂ )-alkylcarbamoyl, N-(C ₃ -C ₈ )-cycloalkylcarbamoyl, N-(C ₆ -C ₁₂ )-arylcarbamoyl, N-(C ₇ -C ₁₆ )-aralkylcarbamoyl, N-(CrC ₁₀ )-alkyl-N-(C ₆ -C ₁₂ )- arylcarbamoyl, N-(CrC ₁₀ )-alkyl-N-(C ₇ -Ci ₆ )-aralkylcarbamoyl, N-((C,-C ₁₀ )-alkoxy-(C ₁ - C ₁₀ )-alkyl)-carbamoyl, N-((C ₆ -C ₁₂ )-aryloxy-(C,-C ₁₀ )-alkyl)-carbamoyl, N-((C ₇ -C ₁₆ )- aralkyloxy-(C ₁ -C ₁₀ )-alkyl)-carbamoyl, N-(C ₁ -Cio)-alkyl-N-((C ₁ -C ₁₀ )-alkoxy-(Ci-C ₁ o)- alkyl)-carbamoyl, N-(C ₁ -Cio)-alkyl-N-((C ₆ -C ₁₂ )-aryloxy-(C ₁ -Cio)-alkyl)-carbamoyl, N- (C ₁ -Cio)-alkyl-N-((C ₇ -C ₁₆ )-aralkyloxy-(C ₁ -C ₁₀ )-alkyl)-carbamoyl, carbamoyloxy, N-(C ₁ - Ci ₂ )-alkylcarbamoyloxy, N.N-di-(C _r Ci ₂ )-alkylcarbamoyloxy, N-(C ₃ -C ₈ )- cycloalkylcarbamoyloxy, N-(C ₆ -C ₁₂ )-arylcarbamoyloxy, N-(C ₇ -C ₁₆ )- aralkylcarbamoyloxy, N-(C ₁ -C ₁₀ )-alkyl-N-(C ₆ -C ₁₂ )-arylcarbamoyloxy, N(Ci -C ₁₀ )-alkyl- N-(C ₇ -Ci ₆ )-aralkylcarbamoyloxy, N-((Ci-Cio)-alkyl)-carbamoyloxy, N-((C ₆ -C ₁₂ )-aryloxy- (Q-QoJ-alky^-carbamoyloxy, N-((C ₇ -C ₁₆ )-aralkyloxy-(C ₁ -C ₁₀ )-alkyl)-carbamoyloxy, N- (C ₁ -C ₁₀ )-alkyl-N-((C ₁ -C ₁₀ )-alkoxy-(C ₁ -C ₁ o)-alkyl)-carbamoyloxy, N-(C ₁ -C ₁ o)-alkyl-N- ((C ₆ -Ci ₂ )-aryloxy-(C ₁ -C ₁₀ )-alkyl)-carbamoyloxy, N-(C ₁ -C ₁₀ )-alkyl-N-((C ₇ -C ₁₆ )- aralkyloxy-(C ₁ -C ₁₀ )-alkyl)-carbamoyloxy, amino, (C _r C ₁₂ )-alkylamino, di-(CrC ₁₂ )- alkylamino, (C ₃ -C ₈ )-cycloalkylamino, (C ₃ -C ₁₂ )-alkenylamino, (C ₃ -C ₁₂ )-alkynylamino, N- (C ₆ -C ₁₂ )-arylamino, N-(C ₇ -C ₁ 0-aralkylamino, N-alkylaralkylamino, N-alkyl-arylamino, (C _r C ₁₂ )-alkoxyamino, (C ₁ -Ci ₂ )-alkoxy-N-(C ₁ -C ₁₀ )-alkylamino, (C ₁ -C ₁₂ )- alkylcarbonylamino, (C ₃ -C ₈ )-cycloalkylcarbonylamino, (C ₆ -C ₁₂ )-arylcarbonylamino, (C ₇ -

C ₁₆ )-alkylcarbonylamino, (C ₁ -C ₁₂ )-alkylcarbonyl-N-(C ₁ -Cio)-alkylamino, (C ₃ -C ₈ )- cycloalkylcarbonyl-N-(C ₁ -C ₁₀ )-alkylamino, (C ₆ -C ₁₂ )-arylcarbonyl-N-(C ₁ -C ₁₀ )- alkylamino, (C ₇ -Ci i)-aralkylcarbonyl-N-(Ci-Ci ₀ )-alkylamino, (C ₁ -C ₁₂ )- alkylcarbonylamino-(Ci-C ₈ )-alkyl, (C ₃ -C ₈ )-cycloalkylcarbonylamino-(Ci-C ₈ )-alkyl, (C ₆ - Ci ₂ )-arylcarbonylamino-(Ci-C ₈ )-alkyl, (C ₇ -Ci ₆ )-aralkylcarbonylamino-(Ci-C ₈ )-alkyl, amino-(C,-C,o)-alkyl, N-(C,-Cio)-alkylamino-(Ci-Cio)alkyl, N.N-di-(C,-C,o)-alkylamino- (C,-Cio)-alkyl, (C ₃ -C ₈ )-cycloalkylamino-(Ci-Ci ₀ )-alkyl, (Ci-Ci ₂ )-alkylmercapto, (C ₁ -

C ₁₂ )-alkylsulfϊnyl, (Ci-Ci ₂ )-alkylsulfonyl, (C ₆ -Ci ₂ )-arylmercapto, (C ₆ -Ci ₂ )-arylsulfϊnyl, (C ₆ -Ci ₂ )-arylsulfonyl, (C ₇ -C i ₆ )-aralkylmercapto, (C ₇ -Ci ₆ )-aralkylsulfinyl, or (C ₇ -C ₁₆ )- aralkylsulfonyl; X is O or S; Q is O, S, NR ¹ , or a bond; where, if Q is a bond, R ⁴ is halogen, nitrile, or trifluoromethyl; or where, if Q is O, S, or NR', R ⁴ is hydrogen, (Ci-Cio)-alkyl radical, (C ₂ -C i _O )-alkenyl radical, (C ₂ -Cio)-alkynyl radical, wherein alkenyl or alkynyl radical contains one or two C-C multiple bonds; unsubstituted fluoroalkyl radical of the formula -[CH ₂ ] _x -C _f H _(2f+1 . _g) -F _g , (C ₁ -C ₈ )-alkoxy-(C ₁ -C ₆ )-alkyl radical, (Ci-C ₆ )-alkoxy-(Ci-C ₄ )-alkoxy-(C ₁ -C ₄ )-alkyl radical, aryl radical, heteroaryl radical, (C ₇ -Ci i)-aralkyl radical, or a radical of the formula Z

-[CH ₂ ] _v -[O] _w -[CH ₂ ] _r E (Z) where E is a heteroaryl radical, a (C ₃ -C ₈ )-cycloalkyl radical, or a phenyl radical of the formula F

v is 0-6, w is 0 or 1, t is 0-3, and R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , and R ¹ ' are identical or different and are hydrogen, halogen, cyano, nitro, trifluoromethyl, (C,-C ₆ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C _r C ₆ )-alkoxy, -O-[CH ₂ ] _X - C _f H _(2f+1 . _g) -F _g , -OCF ₂ -Cl, -0-CF ₂ -CHFCl, (C,-C ₆ )-alkylmercapto, (C ₁ -C ₆ )- hydroxyalkyl, (C ₁ - C ₆ )-alkylsulfinyl, (C,-C ₆ )-alkylsulfonyl, (Ci-C ₆ )-alkylcarbonyl, (C ₁ -C ₈ )- alkoxycarbonyl, carbamoyl, N-(C ₁ -C ₈ )-alkylcarbamoyl, N,N-di-(Ci-C ₈ )- alkylcarbamoyl, or (C ₇ -C ₁ i)-aralkylcarbamoyl, optionally substituted by fluorine, chlorine, bromine, trifluoromethyl, (C]-C ₆ )-alkoxy, N-(C ₃ -C ₈ )- cycloalkylcarbamoyl, N-(C ₃ -C ₈ )-cycloalkyl-(Ci-C ₄ )-alkylcarbamoyl, (C ₁ -C ₆ )- alkylcarbonyloxy, phenyl, benzyl, phenoxy, benzyloxy, NR ^Y R ^Z wherein R ^y and R ^z are independently selected from hydrogen, (Ci-Ci ₂ )-alkyl, (C ₁ -Cg)-BIkOXy-(C ₁ - C ₈ )-alkyl, (C ₇ -C ₁₂ )-aralkoxy-(Ci-C ₈ )-alkyl, (C ₆ -Ci ₂ )-aryloxy-(C,-C ₈ )-alkyl, (C ₃ - C ₁₀ )-cycloalkyl, (C ₃ -C ₁₂ )-alkenyl, (C ₃ -C ₁₂ )-alkynyl, (C ₆ -C ₁₂ )-aryl, (C ₇ -C ₁₁ )- aralkyl, (C _r C ₁₂ )-alkoxy, (C ₇ -C ₁₂ )aralkoxy, (C ₁ -C ₁₂ )-alkylcarbonyl, (C ₃ -C ₈ )- cycloalkylcarbonyl, (C ₆ -C ₁₂ ) arylcarbonyl, (C ₇ -C ₁₆ )-aralkylcarbonyl; or further wherein R ^y and R ² together are -[CH2 ] _h , in which a CH ₂ group can be replaced by O, S, N-(C ₁ -C ₄ )-alkylcarbonylimino, or N-(Ci -C ₄ )-alkoxycarbonylimino; phenylmercapto, phenylsulfonyl, phenylsulfinyl, sulfamoyl, N-(Ci-C ₈ )- alkylsulfamoyl, or N, N-di-(C, -C ₈ )-alkylsulfamoyl; or alternatively R ⁷ and R ⁸ , R ⁸ and R ⁹ , R ⁹ and R ¹⁰ , or R ¹⁰ and R ¹¹ , together are a chain selected from -[CH ₂ ] _n - or -CH=CH-CH=CH-, where a CH ₂ group of the chain is optionally replaced by O, S, SO, SO ₂ , or NR ^Y ; and n is 3, 4, or 5; and if E is a heteroaryl radical, said radical can carry 1-3 substituents selected from those defined for R ⁷ -R ^π , or if E is a cycloalkyl radical, the radical can carry one substituent selected from those defined for R ⁷ -R ^π ; or where, if Q is NR', R ⁴ is alternatively R", where R' and R" are identical or different and are hydrogen, (C ₆ -C _]2 )-aryl, (C ₇ -C ₁ ,)-aralkyl, (C,-C ₈ )-alkyl, (C _r C ₈ )-alkoxy-(Ci-C ₈ )-alkyl, (C ₇ -C ₁₂ )-aralkoxy-(C _r C ₈ )-alkyl, (C ₆ -C ₁₂ )-aryloxy-(C ₁ -C ₈ )-alkyl, (C ₁ -C ₁₀ )-alkylcarbonyl, optionally substituted (C ₇ -C ₁₆ )-aralkylcarbonyl, or optionally substituted C ₆ -C ₁₂ )- arylcarbonyl; or R' and R" together are -[CH ₂ ] _h , in which a CH ₂ group can be replaced by O, S, N-acylimino, or N-(C ₁ -C ₁₀ )-alkoxycarbonylimino, and h is 3 to 7. Y is N or CR ³ ;

R ¹ , R ² and R ³ are identical or different and are hydrogen, hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl, (d-C ₂₀ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C ₃ -C ₈ )cycloalkyl-

(C _r C ₁₂ )-alkyl, (C ₃ -C ₈ )-cycloalkoxy, (C ₃ -Cg)-CyClOaIlCyI-(C ₁ -C ₁₂ )^IkOXy, (C ₃ -C ₈ )- cycloalkyloxy-(C,-C ₁₂ )-alkyl, (C ₃ -C ₈ )-cycloalkyloxy-(C ₁ -C ₁₂ )-alkoxy, (C ₃ -C ₈ )- cycloalkyl^C^C^-alkyl^d-C^-alkoxy, (C ₃ -Cs)-CyClOaIlCyI-(C ₁ -C ₈ )^IkOXy-(Ci-C ₆ )- alkyl, (C ₃ -Cg)-CyClOaHCyIoXy-(C ₁ -Cs)-BIkOXy-(Ci-C ₆ )^IlCyI, (C ₃ -C ₈ )-cycloalkoxy-(C,-C ₈ )- alkoxy-(C _r Cg)-alkoxy, (C ₆ -C ₁₂ )-aryl, (C ₇ -C, ₆ )-aralkyl, (C ₇ -Ci ₆ )-aralkenyl, (C ₇ -C ₁₆ )- aralkynyl, (C ₂ -C ₂₀ )-alkenyl, (C ₂ -C ₂₀ )-alkynyl, (d-C ₂₀ )-alkoxy, (C ₂ -C ₂₀ )-alkenyloxy, (C ₂ - C ₂₀ )-alkynyloxy, retinyloxy, (C ₁ -C ₂₀ )-alkoxy-(C ₁ -C ₁₂ )-alkyl, (C,-C ₁₂ )-alkoxy-(C ₁ -C, ₂ )- alkoxy, (C ₁ -C ₁₂ )-alkoxy-(C ₁ -C ₈ )-alkoxy-(C ₁ -C ₈ )-alkyl, (C ₆ -C ₁₂ )-aryloxy, (C ₇ -C ₁₆ )- aralkyloxy, (C ₆ -C ₁₂ )-aryloxy-(C ₁ -C ₆ )-alkoxy, (C ₇ -C ₁₆ )-aralkoxy-(C ₁ -C ₆ )-alkoxy, (C ₁ -C ₁₆ )- hydroxyalkyl, (C ₆ -Ce)-BIyIoXy-(C ₁ -Cg)-BIlCyI, (C ₇ -C ₁₆ )-aralkoxy-(C ₁ -C ₈ )-alkyl, (C ₆ -C ₁₂ )- aryloxy-(Ci-C ₈ )-alkoxy-(C,-C ₆ )-alkyl, (C ₇ -C ₁₂ )-aralkyloxy-(C,-C ₈ )-alkoxy-(Ci-C ₆ )-alkyl, (C ₂ -C ₂₀ )-alkenyloxy-(C _r C ₆ )-alkyl, (Cz-C^-alkynyloxyKCrC^-alkyl, retinyloxy-(C,- C ₆ )-alkyl, -O-[CH ₂ ] _x CfH _(2f+1 . _g) F _g , -OCF ₂ Cl, -OCF ₂ -CHFCl, (C _r C ₂₀ )-alkylcarbonyl, (C ₃ - C ₈ )-cycloalkylcarbonyl, (C ₆ -C ₁₂ )-arylcarbonyl, (C ₇ -C _]6 )-aralkylcarbonyl, cinnamoyl, (C ₂ - C ₂ o)-alkenylcarbonyl, (C ₂ -C ₂₀ )-alkynylcarbonyl, (C _r C ₂ o)-alkoxycarbonyl, (Ci-C] ₂ )- alkoxy-(Ci-Ci ₂ )-alkoxycarbonyl, (C ₆ -C ₎₂ )-aryloxycarbonyl, (C ₇ -Ci ₆ )-aralkoxycarbonyl, (C ₃ -C ₈ )-cycloalkoxycarbonyl, (C ₂ -C ₂₀ )-alkenyloxycarbonyl, retinyloxycarbonyl, (C ₂ -C ₂₀ )- alkynyloxycarbonyl, (C ₆ -Ci ₂ )-aryloxy-(Ci-C ₆ )-alkoxycarbonyl, (C ₇ -Ci ₆ )-aralkoxy-(Ci- C ₆ )-alkoxycarbonyl, (C ₃ -C ₈ )-cycloalkyl-(Ci -C ₆ )-alkoxycarbonyl, (C ₃ -C ₈ )-cycloalkoxy- (Ci-C ₆ )-alkoxycarbonyl, (Ci-Ci ₂ )-alkylcarbonyloxy, (C ₃ -Cg)-cycloalkylcarbonyloxy, (C ₆ - Ci ₂ )-arylcarbonyloxy, (C ₇ -Ci ₆ )-aralkylcarbonyloxy, cinnamoyloxy, (C ₂ -Ci ₂ )- alkenylcarbonyloxy, (C ₂ -C i ₂ )-alkynylcarbonyloxy, (Ci-C _]2 )-alkoxycarbonyloxy, (Ci-

C ₁₂ )-alkoxy-(C i -C ₁₂ )-alkoxycarbonyloxy, (C ₆ -C ₁₂ )-aryloxycarbonyloxy, (C ₇ -C ₁₆ )- aralkyloxycarbonyloxy, (C ₃ -C ₈ )-cycloalkoxycarbonyloxy, (C ₂ -C] ₂ )- alkenyloxycarbonyloxy, (C ₂ -Ci ₂ )-alkynyloxycarbonyloxy, carbamoyl, N-(Ci-Ci ₂ )- alkylcarbamoyl, N,N-di-(Ci-Ci ₂ )-alkylcarbamoyl, N-(C ₃ -C ₈ )-cycloalkylcarbamoyl, N,N- dicyclo-(C ₃ -C ₈ )-alkylcarbamoyl, N-(Ci-Cio)-alkyl-N-(C ₃ -C ₈ )-cycloalkylcarbamoyl, N-

((C ₃ -C ₈ )-cycloalkyl-(Ci-C ₆ )-alkyl)-carbamoyl, N-(C ₁ -C ₆ )-alkyl-N-((C ₃ -C ₈ )-cycloalkyl- (Ci-C ₆ )-alkyl)-carbamoyl, N-(+)-dehydroabietylcarbamoyl, N-(Ci-C ₆ )-alkyl-N-(+)- dehydroabietylcarbamoyl, N-(C ₆ -Ci ₂ )-arylcarbamoyl, N-(C ₇ -Ci ₆ )-aralkylcarbamoyl, N- (Ci-C ₁₀ )-alkyl-N-(C ₆ -C, ₆ )-arylcarbamoyl, N-(Ci-Cio)-alkyl-N-(C ₇ -Ci ₆ )-aralkylcarbamoyl, N-((Ci-Ci ₈ )-alkoxy-(C ₁ -Cio)-alkyl)-carbamoyl, N-((C ₆ -Ci ₆ )-aryloxy-(Ci-Cio)-alkyl)- carbamoyl, N-((C ₇ -Ci ₆ )-aralkyloxy-(C,-C,o)-alkyl)-carbamoyl, N-(Ci-Cio)-alkyl-N-((C ₁ - C,o)-alkoxy-(Ci-Cio)-alkyl)-carbamoyl, N-(C ₁ -C ₁₀ )-alkyl-N-((C ₆ -C ₁₂ )-aryloxy-(Ci-Cio)- alkyl)-carbamoyl, N-(Ci-Cio)-alkyl-N-((C ₇ -Ci ₆ )-aralkyloxy-(Ci-C ₎₀ )-alkyl)-carbamoyl; CON(CH ₂ ) _h , in which a CH ₂ group can be replaced by O, S, N-(Ci-C ₈ )-alkylimino, N- (C ₃ -C ₈ )-cycloalkylimino, N-(C ₃ -C ₈ )-cycloalkyl-(C,-C ₄ )-alkylimino, N-(C ₆ -Cj ₂ )- arylimino, N-(C ₇ -Ci ₆ )-aralkylimino, N-(Ci-C ₄ )-alkoxy-(Ci-C ₆ )-alkylimino, and h is from 3 to 7; a carbamoyl radical of the formula R

in which R ^x and R ^v are each independently selected from hydrogen, (C,-C ₆ )-alkyl, (C ₃ -C ₇ )- cycloalkyl, aryl, or the substituent of an α-carbon of an oamino acid, to which the L- and D-amino acids belong, s is 1-5, T is OH, or NR*R**, and R*, R** and R*** are identical or different and are selected from hydrogen, (C ₆ -C ₁₂ )-aryl, (C ₇ -C,,)-aralkyl, (Ci-C ₈ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (+)-dehydroabietyl, (C ₁ -C ₈ )-alkoxy-(C ₁ -C ₈ )-alkyl, (C ₇ -Ci ₂ )-aralkoxy-(Ci-C ₈ )- alkyl, (C ₆ -C ₁₂ )-aryloxy-(Ci-C ₈ )-alkyl, (Ci-C ₁₀ )-alkanoyl, optionally substituted (C ₇ -Ci ₆ )-aralkanoyl, optionally substituted (C ₆ -C ₁₂ )-aroyl; or R* and R** together are -[CH ₂ ] _h , in which a CH ₂ group can be replaced by O, S, SO, SO ₂ , N- acylamino, N-(Ci-Ci ₀ )-alkoxycarbonylimino, N-(Ci-C ₈ )-alkylimino, N-(C ₃ -C ₈ )- cycloalkylimino, N-(C ₃ -C ₈ )-cycloalkyl-(Ci -C ₄ )-alkylimino, N-(C ₆ -Ci ₂ )- arylimino, N-(C ₇ -Ci ₆ )-aralkylimino, N-(Ci-C ₄ )-alkoxy-(Ci-C ₆ )-alkylimino, and h carbamoyloxy, N-(C]-Ci ₂ )-alkylcarbamoyloxy, N,N-di-(Ci-Ci ₂ )-alkylcarbamoyloxy, N- (C ₃ -C ₈ )-cycloalkylcarbamoyloxy, N-(C ₆ -Ci ₂ )-arylcarbamoyloxy, N-(C ₇ -C ₁₆ )- aralkylcarbamoyloxy, N-(C ₁ -C ₁₀ )-alkyl-N-(C ₆ -Ci ₂ )-arylcarbamoyloxy, N-(Ci-Cio)-alkyl- N-(C ₇ -C ₁₆ )-aralkylcarbamoyloxy, N-((C ] -C ₁₀ )-alkyl)-carbamoyloxy, N-^C ₆ -C ₁₂ )-aryloxy- (CrC ₁₀ )-alkyl)-carbamoyloxy, N-((C ₇ -C ₁₆ )-aralkyloxy-(Ci-Ci ₀ )-alkyl)-carbamoyloxy, N-

(C _r Cio)-alkyl-N-((C,-C ₁ o)-alkoxy-(C ₁ -Cio)-alkyl)-carbamoyloxy, N-(Ci-C,o)-alkyl-N- ((C ₆ -C ₁₂ )-aryloxy-(Ci-C ₁₀ )-alkyl)-carbamoyloxy, N-(C ₁ -C,o)-alkyl-N-((C ₇ -C ₁₆ )- aralkyloxy-(C i -C ₁₀ )-alkyl)-carbamoyloxyamino, (C]-C ₁₂ )-alkylamino, di-(C i -C ₁₂ )- alkylamino, (C ₃ -C ₈ )-cycloalkylamino, (C ₃ -C ₁₂ )-alkenylamino, (C ₃ -C ₁₂ )-alkynylamino, N- (C ₆ -C ₁₂ )-arylamino, N-(C ₇ -Ci i)-aralkylamino, N-alkyl-aralkylamino, N-alkyl-arylamino,

(Ci-C ₁₂ )-alkoxyamino, (C ₁ -C ₁₂ )-alkoxy-N-(Ci-Ci ₀ )-alkylamino, (C]-Ci ₂ )-alkanoylamino, (C ₃ -C ₈ )-cycloalkanoylamino, (C ₆ -Ci ₂ )-aroylamino, (C ₇ -C] ₆ )-aralkanoylamino, (Ci-C ₁₂ )- alkanoyl-N-(Ci-Cio)-alkylamino, (C ₃ -C ₈ )-cycloalkanoyl-N-(Ci-Ci ₀ )-alkylamino, (C ₆ -Ci ₂ )- aroyl-N-(Ci-Cio)-alkylamino, (C ₇ -C] i)-aralkanoyl-N-(Ci-Ci ₀ )-alkylamino, (Ci-Ci ₂ )- alkanoylamino-(Ci-C ₈ )-alkyl, (C ₃ -C ₈ )-cycloalkanoylamino-(Ci-C ₈ )-alkyl, (C ₆ -C ₁₂ )- aroylamino-(Ci-C ₈ )-alkyl, (C ₇ -C ₁₆ )-aralkanoylamino-(Ci-C ₈ )-alkyl, amino-(Ci-Ci ₀ )-alkyl, N-(C ₁ -C,o)-alkylammo-(Ci-Cio)-alkyl, N,N-di(C ₁ -C,o)-alkylamino-(C,-Cio)-alkyl, (C ₃ - C ₈ )-cycloalkylamino(Ci-Cio)-alkyl, (Ci-C ₂₀ )-alkylmercapto, (Ci-C ₂₀ )-alkylsulfϊnyl, (Ci- C ₂₀ )-alkylsulfonyl, (C ₆ -Ci ₂ )-arylmercapto, (C ₆ -Ci ₂ )-arylsulfinyl, (C ₆ -Ci ₂ )-arylsulfonyl, (C ₇ -C, ₆ )-aralkylmercapto, (C ₇ -Ci ₆ )-aralkylsulfinyl, (C ₇ -C, ₆ )-aralkylsulfonyl, (C,-Ci ₂ )- alkylmercapto-(C,-C ₆ )-alkyl, (C,-C ₁₂ )-alkylsulfinyl-(Ci-C ₆ )-alkyl, (C,-Ci ₂ )-alkylsulfonyl- (Ci-C ₆ )-alkyl, (C ₆ -Ci ₂ )-arylmercapto-(Ci-C ₆ )-alkyl, (C ₆ -C ₁₂ )-arylsulfinyl-(Ci-C ₆ )-alkyl, (C ₆ -C, ₂ )-arylsulfonyl-(Ci-C ₆ )-alkyl, (C ₇ -Ci ₆ )-aralkylmercapto-(Ci-C ₆ )-alkyl, (C ₇ -Ci ₆ )- aralkylsulfinyl-(C,-C ₆ )-alkyl, (C ₇ -Ci ₆ )-aralkylsulfonyl-(C ₁ -C ₆ )-alkyl, sulfamoyl, N-(C ₁ - C,o)-alkylsulfamoyl, N,N-di-(Ci-C, ₀ )-alkylsulfamoyl, (C ₃ -C ₈ )-cycloalkylsulfamoyl, N-

(C ₆ -C ₁₂ )-arylsulfamoyl, N-(C ₇ -C, ₆ )-aralkylsulfamoyl, N-(C,-C,o)-alkyl-N-(C ₆ -C ₁₂ )- arylsulfamoyl, N-(C, -Cio)-alkyl-N-(C ₇ -Ci ₆ )-aralkylsulfamoyl, (Ci -Cio)-alkylsulfonamido, N-((Ci-C ₁₀ )-alkyl)-(Ci-Cio)-alkylsulfonamido, (C ₇ -Ci ₆ )-aralkylsulfonamido, and N-((C,- Cio)-alkyl-(C ₇ -Ci ₆ )-aralkylsulfonamido; where an aryl radical may be substituted by 1 to 5 substituents selected from hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl, (C ₂ -C ₁₆ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C ₃ -C ₈ )-cycloalkyl-(Ci-C ₁₂ )-alkyl, (C ₃ -C ₈ )- cycloalkoxy, (C ₃ -C ₈ )-cycloalkyl-(C,-C ₁₂ )-alkoxy, (C ₃ -C ₈ )-cycloalkyloxy-(C ₁ -Ci ₂ )-alkyl,

(C ₃ -C ₈ )-cycloalkyloxy-(Ci-Ci ₂ )-alkoxy, (C ₃ -C ₈ )-cycloalkyl-(Ci-C ₈ )-alkyl-(Ci-C ₆ )-alkoxy, (C ₃ -C ₈ )-cycloalkyl(Ci-C ₈ )-alkoxy-(Ci-C ₆ )-alkyl, (C ₃ -Cs)-CyClOaHCyIoXy-(C ₁ -Cs)-BIkOXy- (C,-C ₆ )-alkyl, (Q-CsVcycloalkoxy-tCrCjO-alkoxyKQ-CsO-alkoxy, (C ₆ -C ₁₂ )-aryl, (C ₇ - C ₁₆ )-aralkyl, (C ₂ -C ₁₆ )-alkenyl, (C ₂ -C ₁₂ )-alkynyl, (C,-C ₁₆ )-alkoxy, (C ₁ -C ₁₆ )-alkenyloxy, (C ₁ -C ₁₂ )-alkoxy-(C ₁ -C ₁₂ )-alkyl, (C ₁ -C ₁₂ )-alkoxy-(C ₁ -C ₁₂ )-alkoxy, alkoxy-(C,-Cs)-alkyl, (C ₆ -C ₁₂ )-aryloxy, (C ₇ -C ₁₆ )-aralkyloxy, (C ₆ -C ₁₂ )-aryloxy-(C _r C ₆ )- alkoxy, (C ₇ -C ₁₆ )-aralkoxy-(d-C ₆ )-alkoxy, (Ci-C ₈ )-hydroxyalkyl, (C ₆ -C ₁₆ )-aryloxy-(C ₁ - C ₈ )-alkyl, (C ₇ -C ₁₆ )-aralkoxy-(C ₁ -C ₈ )-alkyl, (C ₆ -C ₁₂ )^rYlOXy-(C ₁ -Cs)-BIkOXy-(C ₁ -C ₆ )- alkyl, (C.-C^-aralkyloxy-tCi-CsO-alkoxy-tC-C^-alkyl, -O-[CH ₂ ] _x C _f H _(2f+1 . _g) F _g , -OCF ₂ Cl, -OCF ₂ -CHFCl, (C _r C ₁₂ )-alkylcarbonyl, (C ₃ -C ₈ )-cycloalkylcarbonyl, (C ₆ -C ₁₂ )- arylcarbonyl, (C ₇ -C ₁₆ )-aralkylcarbonyl, (C ₁ -C ₁₂ )-alkoxycarbonyl, (Ci-C ₁₂ )-alkoxy-(C _! - C ₁₂ )-alkoxycarbonyl, (C ₆ -C ₁₂ )-aryloxycarbonyl, (C ₇ -C ₁₆ )-aralkoxycarbonyl, (C ₃ -C ₈ )- cycloalkoxycarbonyl, (C ₂ -C ₁₂ )-alkenyloxycarbonyl, (C ₂ -Ci ₂ )-alkynyloxycarbonyl, (C ₆ - C ₁₂ )-aryloxy-(C ₁ -C ₆ )-alkoxycarbonyl, (C ₇ -C ₁₆ )-aralkoxy-(C _! -C ₆ )-alkoxycarbonyl, (C ₃ - C ₈ )-cycloalkyl-(C ₁ -C ₆ )-alkoxycarbonyl, (C ₃ -Cg)-cycloalkoxy-(C _r C ₆ )-alkoxycarbonyl,

(C ₁ -C ₁₂ )-alkylcarbonyloxy, (C ₃ -C ₈ )-cycloalkylcarbonyloxy, (C ₆ -C ₁₂ )-arylcarbonyloxy, (C ₇ -C ₁₆ )-aralkylcarbonyloxy, cinnamoyloxy, (C ₂ -C ₁₂ )-alkenylcarbonyloxy, (C ₂ -Ci ₂ )- alkynylcarbonyloxy, (C ] -C ₁₂ )-alkoxycarbonyloxy, (C ] -C ] ₂ )-alkoxy-(C ₁ -Ci ₂ )- alkoxycarbonyloxy, (C ₆ -C] ₂ )-aryloxycarbonyloxy, (C ₇ -C ₁₆ )-aralkyloxycarbonyloxy, (C ₃ - C ₈ )-cycloalkoxycarbonyloxy, (C ₂ -Ci ₂ )-alkenyloxycarbonyloxy, (C ₂ -Ci ₂ )- alkynyloxycarbonyloxy, carbamoyl, N-(Ci-Ci ₂ )-alkylcarbamoyl, N,N-di(C _r C ₁₂ )- alkylcarbamoyl, N-(C ₃ -C ₈ )-cycloalkylcarbamoyl, N,N-dicyclo-(C ₃ -C ₈ )-alkylcarbamoyl, N-(Ci-C ₁₀ )-alkyl-N-(C ₃ -C ₈ )-cycloalkylcarbamoyl, NK(C ₃ -Cs)-CyClOaIlCyI-(C ₁ -C ₆ )- alkyl)carbamoyl, N-(C ₁ -C ₆ )-alkyl-N-((C ₃ -C ₈ )-cycloalkyl-(C ₁ -C ₆ )-alkyl)carbamoyl, N-(+)- dehydroabietylcarbamoyl, N-(Ci-C ₆ )-alkyl-N-(+)-dehydroabietylcarbamoyl, N-(C ₆ -C ₁₂ )- arylcarbamoyl, N-(C ₇ -C ₁₆ )-aralkylcarbamoyl, N-(C ₁ -C ₁ O)-BIlCyI-N-(C ₆ -C ₁₆ )- arylcarbamoyl, N-(C,-C ₁₀ )-alkyl-N-(C ₇ -C ₁₆ )-aralkylcarbamoyl, C ₁₀ )-alkyl)carbamoyl, N-((C ₆ -C ₁₆ )-aryloxy-(C ₁ -C ₁₀ )-alkyl)carbamoyl, N-((C ₇ -C ₁₆ )- aralkyloxy-(C ₁ -C ₁₀ )-alkyl)carbamoyl, N-(C ₁ -C ₁₀ )-alkyl-N-((C ₁ -C ₁₀ )-alkoxy-(C ₁ -C ₁₀ )- alkyl)carbamoyl, N-(C ₁ -C ₁₀ )-alkyl-N-((C ₆ -C, ₂ )-aryloxy-(Ci-C ₁₀ )-alkyl)carbamoyl, N-(C ₁ -

C ₁₀ )-alkyl-N-((C ₇ -C ₁₆ )-aralkyloxy-(C ₁ -C ₁₀ )-alkyl)-carbamoyl, CON(CH ₂ ) _h , in which a CH ₂ group can be replaced by, O, S, N-(C ₁ -C ₈ )-alkylimino, N-(C ₃ -C ₈ )-cycloalkylimino, N-(C ₃ -C ₈ )-cycloalkyl-(Ci-C4)-alkylimino, N-(C ₆ -C ₁₂ )-arylimino, N-(C ₇ -C ₁₆ )- aralkylimino, N-(Ci-C ₄ )-alkoxy-(Ci-C ₆ )-alkylimino, and h is from 3 to 7; carbamoyloxy, N-(C ₁ -Ci ₂ )-alkylcarbamoyloxy, N,N-di-(Ci-C, ₂ )-alkylcarbamoyloxy, N-(C ₃ -C ₈ )- cycloalkylcarbamoyloxy, N-(C ₆ -C i ₆ )-arylcarbamoyloxy, N-(C ₇ -C ₁₆ )- aralkylcarbamoyloxy, N-(Ci-C ₁₀ )-alkyl-N-(C ₆ -Ci ₂ )-arylcarbamoyloxy, N-(Ci-C ₁₀ )-alkyl-

N-(C ₇ -Ci ₆ )-aralkylcarbamoyloxy, N-((Ci-Cio)-alkyl)carbamoyloxy, N-((C ₆ -Ci ₂ )-aryloxy- (C ₁ -Cio)-alkyl)carbamoyloxy, N-((C ₇ -Ci ₆ )-aralkyloxy-(Ci-Cio)-alkyl)carbamoyloxy, N- (Ci-C ₁ o)-alkyl-N-((C,-Cio)-alkoxy-(Ci-C ₁₀ )-alkyl)carbamoyloxy, N-(Ci-C ₁₀ )-alkyl-N- ((C ₆ -C ₁₂ )-aryloxy-(C,-C ₁ o)-alkyl)carbamoyloxy, N-(C ₁ -C ₁₀ )-alkyl-N-((C ₇ -C ₁₆ )- aralkyloxy-(C ₁ -Cio)-alkyl)carbamoyloxy, amino, (Ci-C ₁₂ )-alkylamino, di-(Ci-C ₁₂ )- alkylamino, (C ₃ -C ₈ )-cycloalkylamino, (C ₃ -Ci ₂ )-alkenylamino, (C ₃ -Ci ₂ )-alkynylamino, N- (C ₆ -Ci ₂ )-arylamino, N-(C ₇ -Ci i)-aralkylamino, N-alkyl-aralkylamino, N-alkyl-arylamino, (Ci-Ci ₂ )-alkoxyamino, (Ci-Ci ₂ )-alkoxy-N-(Ci-Ci ₀ )-alkylamino, (C]-Ci ₂ )-alkanoylamino, (C ₃ -C ₈ )-cycloalkanoylamino, (C ₆ -Ci ₂ )-aroylamino, (C ₇ -C ₁₆ )-aralkanoylamino, (Ci-Ci ₂ )- alkanoyl-N-(C ₁ -C ₁₀ )-alkylamino, (C ₃ -C ₈ )-cycloalkanoyl-N-(C,-Ci ₀ )-alkylamino, (C ₆ -Ci ₂ )- aroyl-N-(Ci-Cio)-alkylamino, (C ₇ -Ci i)-aralkanoyl-N-(C ₁ -Ci ₀ )-alkylamino, (Ci-C ₁₂ )- alkanoylamino-(Ci-C ₈ )-alkyl, (C ₃ -C ₈ )-cycloalkanoylamino-(Ci-C ₈ )-alkyl, (C ₆ -C ₁₂ )- aroylamino- (Ci-C ₈ )-alkyl, (C ₇ -Ci ₆ )-aralkanoylamino-(Ci-C ₈ )-alkyl, amino-(Ci-Cio)- alkyl, N-(Ci-C,o)-alkylamino-(Ci-Cio)-alkyl, N,N-di-(C ₁ -Cio)-alkylamino-(Ci-Cio)-alkyl, (C ₃ -C ₈ )-cycloalkylamino-(Ci-Cio)-alkyl, (Ci-Ci ₂ )-alkylmercapto, (C _r Ci ₂ )-alkylsulfinyl,

(Ci-Ci ₂ )-alkylsulfonyl, (C ₆ -Ci ₆ )-arylmercapto, (C ₆ -C _]6 )-arylsulfinyl, (C ₆ -Ci ₆ )- arylsulfonyl, (C ₇ -C i ₆ )-aralkylmercapto, (C ₇ -Ci ₆ )-aralkylsulfinyl, or (C ₇ -Ci ₆ )- aralkylsulfonyl; or wherein R ¹ and R ² , or R ² and R ³ form a chain [CH ₂ ] ₀ , which is saturated or unsaturated by a C=C double bond, in which 1 or 2 CH ₂ groups are optionally replaced by O, S, SO, SO ₂ , or NR ¹ , and R ¹ is hydrogen, (C ₆ -C _]2 )-aryl, (Ci-C ₈ )-alkyl, (d-C ₈ )-alkoxy-(Ci-C ₈ )-alkyl, (C ₇ -C, ₂ )-aralkoxy-(Ci -C ₈ )-alkyl, (C ₆ -C, ₂ )-aryloxy-(C, -C ₈ )-alkyl, (C ₁ -C _]0 )-alkanoyl, optionally substituted (C ₇ -Ci ₆ )-aralkanoyl, or optionally substituted (C6-C12)-aroyl; and o is 3, 4 or 5; or wherein the radicals R ¹ and R ² , or R ² and R ³ , together with the pyridine or pyridazine carrying them, form a 5,6,7,8-tetrahydroisoquinoline ring, a 5,6,7,8-tetrahydroquinoline ring, or a 5,6,7,8-tetrahydrocinnoline ring; or wherein R ¹ and R ² , or R ² and R ³ form a carbocyclic or heterocyclic 5- or 6-membered aromatic ring; or where R ¹ and R ² , or R ² and R ³ , together with the pyridine or pyridazine carrying them, form an optionally substituted heterocyclic ring systems selected from thienopyridines, furanopyridines, pyridopyridines, pyrimidinopyridines, imidazopyridines, thiazolopyridines, oxazolopyridines, quinoline, isoquinoline, and cinnoline; where quinoline, isoquinoline or cinnoline preferably satisfy the formulae Ia, Ib and Ic:

and the substituents R ¹² to R ²³ in each case independently of each other have the meaning ofR ¹ , R ² and R ³ ; or wherein the radicals R ¹ and R ² , together with the pyridine carrying them, form a compound of Formula Id:

where V is S, O, or NR ^k , and R ^k is selected from hydrogen, (Ci-C ₆ )-alkyl, aryl, or benzyl; where an aryl radical may be optionally substituted by 1 to 5 substituents as defined above; and R ²⁴ , R ²⁵ , R ²⁶ , and R ²⁷ in each case independently of each other have the meaning of R ¹ , R ² and R ³ ; f is l to 8; g is 0 or 1 to (2f+l); x is 0 to 3; and h is 3 to 7; including the physiologically active salts and prodrugs derived therefrom.

Compounds of Formulae (I), (Ia), (Ib), (Ic), and (Id) are representative of the heterocyclic carboxamides identified, supra, as being suitable for use in the present invention, Exemplary compounds according to Formula (I) are described in European Patent Nos. EP0650960 and EP0650961. All compounds listed in EP0650960 and EP0650961, in particular, those listed in the compound claims and the final products of the working examples, are hereby incorporated into the present application by reference herein. Additionally, exemplary compounds according to Formula (I) are described in U.S. Patent No. 5,658,933. All compounds listed in U.S. Patent No. 5,658,933, in particular, those listed in the compound claims and the final products of the working examples, are hereby incorporated into the present application by reference herein.

Additional compounds according to Formula (I) are substituted heterocyclic carboxyamides described in U.S. Patent No. 5,620,995; 3-hydroxypyridine-2-carboxamidoesters described in U.S. Patent No. 6,020,350; sulfonamidocarbonylpyridine-2-carboxamides described in U.S. Patent No. 5,607,954; and sulfonamidocarbonyl-pyridine-2-carboxamides and sulfonamidocarbonyl-pyridine-2-carboxamide esters described in U.S. Patent Nos. 5,610,172 and 5,620,996. All compounds listed in these patents, in particular, those compounds listed in the compound claims and the final products of the working examples, are hereby incorporated into the present application by reference herein.

Exemplary compounds according to Formula (Ia) are described in U.S. Patent Nos. 5,719,164 and

5,726,305. All compounds listed in the foregoing patents, in particular, those listed in the compound claims and the final products of the working examples, are hereby incorporated into the present application by reference herein.

As discussed, supra, compounds according the the present invention are in some embodiments heterocyclic carboxamides; in particular, quinoline carboxamides. In certain embodiments, compounds for use in the invention are quinoline-2 -carboxamides. In one embodiment, the compound is selected from a compound of the Formula Ia wherein

A is -CR ⁵ R ⁶ -, and R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, (Ci-C ₆ )-alkyl, (C ₃ -C ₇ )-cycloalkyl, aryl, or a substituent of the α-carbon atom of an α-amino acid, wherein the amino acid is a natural L-amino acid or its D-isomer;

B is -CO ₂ H or a CO ₂ -G carboxyl radical, where G is a radical of an alcohol G-OH in which G is selected from the group consisting of (Ci-C ₂₀ )-alkyl radical, (C ₃ -C ₈ ) cycloalkyl radical, (C ₂ -C ₂₀ )- alkenyl radical, (C ₃ -C ₈ )-cycloalkenyl radical, retinyl radical, (C ₂ -C ₂₀ )-alkynyl radical, (C ₄ -C ₂₀ )- alkenynyl radical;

X is O; Q is O;

R ⁴ is selected from the group consisting of hydrogen, (Ci-Ci _O )-alkyl, (C ₂ -C _]0 )-alkenyl, (C ₂ -C ₁₀ )-alkynyl, wherein alkenyl or alkynyl contains one or two C-C multiple bonds; unsubstituted fluoroalkyl radical of the formula -[CH ₂ ] _x -C _f H _(2f+ i. _g) -F _g , aryl, heteroaryl, and (C ₇ -Ci i)-aralkyl;

R ¹ , R ¹² , R ¹³ , R ¹⁴ and R ¹⁵ are identical or different and are selected from the group consisting of hydrogen, hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl; (Ci-C ₂₀ )-alkyl, (C ₃ -C ₈ )- cycloalkyl, (C ₃ -C ₈ )-cycloalkoxy, (C ₆ -C, ₂ )-aryl, (C ₇ -Ci ₆ )-aralkyl, (C ₇ -Ci ₆ )-aralkenyl, (C ₇ -Ci ₆ )- aralkynyl, (C ₂ -C ₂₀ )-alkenyl, (C ₂ -C ₂₀ )-alkynyl, (C,-C ₂₀ )-alkoxy, (C ₂ -C ₂₀ )-alkenyloxy, (C ₂ -C ₂₀ )- alkynyloxy, retinyloxy, (C ₆ -C i ₂ )-aryloxy, (C ₇ -Ci ₆ )-aralkyloxy, (Ci-Ci ₆ )-hydroxyalkyl, -O- [CH ₂ ] _x CiH _(2f+ ,. _g) F _g , -OCF ₂ Cl, -OCF ₂ -CHFCl, (C ₁ -C ₂₀ )-alkylcarbonyl, (C ₃ -C ₈ )-cycloalkylcarbonyl, (C ₆ -Ci ₂ )-arylcarbonyl, (C ₇ -C ₁₆ )-aralkylcarbonyl, cinnamoyl, (C ₂ -C ₂₀ )-alkenylcarbonyl, (C ₂ -C ₂₀ )- alkynylcarbonyl, (Ci-C ₂₀ )-alkoxycarbonyl, (C ₆ -Ci ₂ )-aryloxycarbonyl, (C ₇ -Ci ₆ )-aralkoxycarbonyl, (C ₃ -C ₈ )-cycloalkoxycarbonyl, (C ₂ -C ₂₀ )-alkenyloxycarbonyl, retinyloxycarbonyl, (C ₂ -C ₂₀ )- alkynyloxycarbonyl, (Ci-C ₁₂ )-alkylcarbonyloxy, (C ₃ -C ₈ )-cycloalkylcarbonyloxy, (C ₆ -Ci ₂ )- arylcarbonyloxy, (C ₇ -Ci ₆ )-aralkylcarbonyloxy, cinnamoyloxy, (C ₂ -Ci ₂ )-alkenylcarbonyloxy, (C ₂ - Ci ₂ )-alkynylcarbonyloxy, (Ci-Ci ₂ )-alkoxycarbonyloxy, (C ₆ -Ci ₂ )-aryloxycarbonyloxy, (C ₇ -Ci ₆ )- aralkyloxycarbonyloxy, (C ₃ -C ₈ )-cycloalkoxycarbonyloxy, (C ₂ -Ci ₂ )-alkenyloxycarbonyloxy, (C ₂ -Ci ₂ )- alkynyloxycarbonyloxy, carbamoyl, N-(Cj -C _]2 )-alkylcarbamoyl, N,N-di-(Ci-Ci ₂ )-alkylcarbamoyl, N- (C ₃ -C ₈ )-cycloalkylcarbamoyl, N,N-dicyclo-(C ₃ -C ₈ )-alkylcarbamoyl, N-(C,-Cio)-alkyl-N-(C ₃ -C ₈ )- cycloalkylcarbamoyl, N-((C ₃ -C ₈ )-cycloalkyl-(Ci -C ₆ )-alkyl)-carbamoyl, N-(+)- dehydroabietylcarbamoyl, N-(Ci-C ₆ )-alkyl-N-(+)-dehydroabietylcarbamoyl, N-(C ₆ -Ci ₂ )- arylcarbamoyl, N-(C ₇ -C] ₆ )-aralkylcarbamoyl, N-(Ci-Ci ₀ )-alkyl-N-(C ₆ -Ci ₆ )-arylcarbamoyl, N-(C ₁ - Cio)-alkyl-N-(C ₇ -Ci ₆ )-aralkylcarbamoyl, carbamoyloxy, N-(Ci-Ci ₂ )-alkylcarbamoyloxy, N,N-di-(Ci- C ₁₂ )-alkylcarbamoyloxy, N-(C ₃ -C ₈ )-cycloalkylcarbamoyloxy, N-(C ₆ -Ci ₂ )-arylcarbamoyloxy, N-(C ₇ - Ci ₆ )-aralkylcarbamoyloxy, N-(Ci-Ci ₀ )-alkyl-N-(C ₆ -Ci ₂ )-arylcarbamoyloxy, N-(C ₁ -C ₁₀ )-alkyl-N-(C ₇ - Ci ₆ )-aralkylcarbamoyloxy, N-((C _r Cio)-alkyl)-carbamoyloxy, N-(Ci-Ci _O )-alkyl-N-((C ₇ -Ci ₆ )- aralkyloxy-(Ci-Cio)-alkyl)-carbamoyloxyamino, (Ci-Ci ₂ )-alkylamino, di-(Ci-C _]2 )-alkylamino, (C ₃ - C ₈ )-cycloalkylamino, (C ₃ -Ci ₂ )-alkenylamino, (C ₃ -Ci ₂ )-alkynylamino, N-(C ₆ -Ci ₂ )-arylamino, N-(C ₇ - Cii)-aralkylamino, N-alkyl-aralkylamino, N-alkyl-arylamino, (Ci-Ci ₂ )-alkoxyamino, (C ₁ -C ₁₂ )- alkoxy-N-(Ci-Ci ₀ )-alkylamino, (Ci-Ci ₂ )-alkanoylamino, (C ₃ -C ₈ )-cycloalkanoylamino, (C ₆ -C] ₂ )- aroylamino, (C ₇ -Ci ₆ )-aralkanoylamino, (Ci-Ci ₂ )-alkanoyl-N-(C ₁ -C ₁₀ )-alkylamino, (C ₃ -C ₈ )- cycloalkanoyl-N-(Ci-Ci ₀ )-alkylamino, (C ₆ -Ci ₂ )-aroyl-N-(Ci-Cio)-alkylamino, (C ₇ -Ci O-aralkanoyl-N- (Ci-C ₁₀ )-alkylamino, amino-(Ci-Ci ₀ )-alkyl, (Ci-C ₂₀ )-alkylmercapto, (Ci-C ₂₀ )-alkylsulfinyl, (C _r C ₂₀ )- alkylsulfonyl, (C ₆ -Ci ₂ )-arylmercapto, (C ₆ -Ci ₂ )-arylsulfinyl, (C ₆ -Ci ₂ )-arylsulfonyl, (C ₇ -Ci ₆ )- aralkylmercapto, (C ₇ -Ci ₆ )-aralkylsulfϊnyl, (C ₇ -Ci ₆ )-aralkylsulfonyl, sulfamoyl, N-(C ₁ -Ci ₀ )- alkylsulfamoyl, N,N-di-(Ci-C, ₀ )-alkylsulfamoyl, (C ₃ -C ₈ )-cycloalkylsulfamoyl, N-(C ₆ -C ₁₂ )- arylsulfamoyl, N-(C ₇ -Ci ₆ )-aralkylsulfamoyl, N-(Ci-Ci ₀ )-alkyl-N-(C ₆ -Ci ₂ )-arylsulfamoyl, N-(Ci-C ₁₀ )- alkyl-N-(C ₇ -Ci ₆ )-aralkylsulfamoyl, (Ci-Cio)-alkylsulfonamido, (C ₇ -Ci ₆ )-aralkylsulfonamido, and N- ((Ci-Cio)-alkyl-(C ₇ -Ci ₆ )-aralkylsulfonamido; where an aryl radical may be substituted by 1 to 5 substituents selected from hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl, (C ₂ -Ci ₆ )-alkyl, (C ₃ -Cg)-cycloalkyl, (C ₃ -C ₈ )-cycloalkoxy, (C ₆ -C, ₂ )-aryl, (C ₇ -C, ₆ )-aralkyl, (C ₂ -Ci ₆ )-alkenyl, (C ₂ -C ₁₂ )- alkynyl, (Ci-C, ₆ )-alkoxy, (C,-C, ₆ )-alkenyloxy, (C ₆ -C i ₂ )-aryloxy, (C ₇ -C, ₆ )-aralkyloxy, (Ci-C ₈ )- hydroxyalkyl, -O-fCH^C _f He _M . _gj F _g , -OCF ₂ Cl, and -OCF ₂ -CHFCl; x is 0 to 3; f is 1 to 8; and g is 0 or 1 to (2f+l); including the physiologically active salts, esters, and prodrugs derived therefrom.

In certain embodiments, the quinoline-2-carboxamide is selected from a compound of the Formula Ia wherein

A is -CHR ⁵ and R ⁵ is hydrogen or methyl; B is -CO ₂ H; X is O;

Q is O;

R ⁴ is hydrogen; and R ¹ , R ¹² , R ¹³ , R ¹⁴ and R ¹⁵ are identical or different and are selected from the group consisting hydrogen, chloro, aryl, aryloxy, and substituted aryloxy, including the physiologically active salts, esters, and prodrugs derived therefrom.

Exemplary compounds according to Formula (Ib) are described in U.S. Patent No. 6,093,730. All compounds listed in U.S. Patent No. 6,093,730, in particular, those listed in the compound claims and the final products of the working examples, are hereby incorporated into the present application by reference herein.

As discussed previously, compounds according to the present invention include isoquinoloine caarboxamides. In certain embodiments, compounds for use in the invention are isoquinoline-3- carboxamides. In one embodiment, the isoquinoline-3-carboxamide is selected from a compound of the Formula Ib wherein

A is -CR ⁵ R ⁶ -, and R ⁵ and R ⁶ are each independently selected from the group consisting of hydrogen, (Ci-C ₆ )-alkyl, (C ₃ -C ₇ )-cycloalkyl, aryl, or a substituent of the ot-carbon atom of an α-amino acid, wherein the amino acid is a natural L-amino acid or its D-isomer;

B is -CO ₂ H or a CO ₂ -G carboxyl radical, where G is a radical of an alcohol G-OH in which G is selected from the group consisting of (Ci-C ₂₀ )-alkyl radical, (C ₃ -C ₈ ) cycloalkyl radical, (C ₂ -C ₂₀ )- alkenyl radical, (C ₃ -C ₈ )-cycloalkenyl radical, retinyl radical, (C ₂ -C ₂ o)-alkynyl radical, (C ₄ -C ₂₀ )- alkenynyl radical; X is O; Q is O; R ⁴ is selected from the group consisting of hydrogen, (Ci-Cio)-alkyl, (C ₂ -Ci ₀ )-alkenyl, (C ₂ -Ci ₀ )-alkynyl, wherein alkenyl or alkynyl contains one or two C-C multiple bonds; unsubstituted fluoroalkyl radical of the formula -[CH ₂ ] _x -C _f H _(2f+1 . _g) -F _g , aryl, heteroaryl, and (C ₇ -Ci i)-aralkyl; R ³ , R ¹⁶ , R ¹⁷ , R ¹⁸ and R ¹⁹ are identical or different and are selected from the group consisting of hydrogen, hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl; (C _r C ₂ o)-alkyl, (C ₃ -C ₈ )- cycloalkyl, (C ₃ -C ₈ )-cycloalkoxy, (C ₆ -Ci ₂ )-aryl, (C ₇ -C ₁₆ )-aralkyl, (C ₇ -C ₁₆ )-aralkenyl, (C ₇ -C ₁₆ )- aralkynyl, (C ₂ -C ₂₀ )-alkenyl, (C ₂ -C ₂₀ )-alkynyl, (C _r C ₂₀ )-alkoxy, (C ₂ -C ₂₀ )-alkenyloxy, (C ₂ -C ₂₀ )- alkynyloxy, retinyloxy, (C ₆ -C ₁₂ )-aryloxy, (C ₇ -C i ₆ )-aralkyloxy, (Ci-Ci ₆ )-hydroxyalkyl, -O-

[CH ₂ ] _x CfH _(2f+1 . _g) F _g , -OCF ₂ Cl, -OCF ₂ -CHFCl, (C ₁ -C ₂₀ )-alkylcarbonyl, (C ₃ -C ₈ )-cycloalkylcarbonyl, (C ₆ -Ci ₂ )-arylcarbonyl, (C ₇ -Ci ₆ )-aralkylcarbonyl, cinnamoyl, (C ₂ -C ₂₀ )-alkenylcarbonyl, (C ₂ -C ₂₀ )- alkynylcarbonyl, (C _r C ₂₀ )-alkoxycarbonyl, (C ₆ -C ₁₂ )-aryloxycarbonyl, (C ₇ -Ci ₆ )-aralkoxycarbonyl, (C ₃ -C ₈ )-cycloalkoxycarbonyl, (C ₂ -C ₂₀ )-alkenyloxycarbonyl, retinyloxycarbonyl, (C ₂ -C ₂₀ )- alkynyloxycarbonyl, (Ci-Ci ₂ )-alkylcarbonyloxy, (C ₃ -C ₈ )-cycloalkylcarbonyloxy, (C ₆ -C ₁₂ )- arylcarbonyloxy, (C ₇ -C i ₆ )-aralkylcarbonyloxy, cinnamoyloxy, (C ₂ -Ci ₂ )-alkenylcarbonyloxy, (C ₂ - Ci ₂ )-alkynylcarbonyloxy, (d-C ₁₂ )-alkoxycarbonyloxy, (C ₆ -Ci ₂ )-aryloxycarbonyloxy, (C ₇ -C ₁₆ )- aralkyloxycarbonyloxy, (C ₃ -C ₈ )-cycloalkoxycarbonyloxy, (C ₂ -C ₁₂ )-alkenyloxycarbonyloxy, (C ₂ -C ₁₂ )- alkynyloxycarbonyloxy, carbamoyl, N-(C ₁ -C ₁₂ )-alkylcarbamoyl, N,N-di-(C ₁ -C ₁₂ )-alkylcarbamoyl, N- (C ₃ -C ₈ )-cycloalkylcarbamoyl, N,N-dicyclo-(C ₃ -C ₈ )-alkylcarbamoyl, N-(C _r C ₁₀ )-alkyl-N-(C ₃ -C ₈ )- cycloalkylcarbamoyl, N-((C ₃ -C ₈ )-cycloalkyl-(C _! -C ₆ )-alkyl)-carbamoyl, N-(+)- dehydroabietylcarbamoyl, N-(C ₁ -C ₆ )-alkyl-N-(+)-dehydroabietylcarbamoyl, N-(C ₆ -Q ₂ )- arylcarbamoyl, N-(C ₇ -C ₁₆ )-aralkylcarbamoyl, N-(C ₁ -C ₁₀ )-alkyl-N-(C ₆ -C ₁₆ )-arylcarbamoyl, N-(C ₁ - Cio)-alkyl-N-(C ₇ -Ci ₆ )-aralkylcarbamoyl, carbamoyloxy, N-(C ₁ -C ₁₂ )-alkylcarbamoyloxy, N,N-di-(Q- Ci ₂ )-alkylcarbamoyloxy, N-(C ₃ -C ₈ )-cycloalkylcarbamoyloxy, N-(C ₆ -C ₁₂ )-arylcarbamoyloxy, N-(C ₇ - c ₁₆ )-aralkylcarbamoyloxy, N-(Ci -Ci ₀ )-alkyl-N-(C ₆ -C i ₂ )-arylcarbamoyloxy, N-(C i -C i _O )-alkyl-N-(C ₇ - C ₁₆ )-aralkylcarbamoyloxy, N-((C, -Ci ₀ )-alkyl)-carbamoyloxy, N-(Q -Q ₀ )-alkyl-N-((C ₇ -Ci ₆ )- aralkyloxy-(Ci-Ci ₀ )-alkyl)-carbamoyloxyamino, (C]-Ci ₂ )-alkylamino, di-(Ci-Ci ₂ )-alkylamino, (C ₃ - C ₈ )-cycloalkylamino, (C ₃ -Ci ₂ )-alkenylamino, (C ₃ -Ci ₂ )-alkynylamino, N-(C ₆ -Ci ₂ )-arylamino, N-(C ₇ - C _u )-aralkylamino, N-alkyl-aralkylamino, N-alkyl-arylamino, (C _r Ci ₂ )-alkoxyamino, (Ci-C] ₂ )- alkoxy-N-(Ci-Cio)-alkylamino, (C _r Ci ₂ )-alkanoylamino, (C ₃ -C ₈ )-cycloalkanoylamino, (C ₆ -Ci ₂ )- aroylamino, (C ₇ -Ci ₆ )-aralkanoylamino, (Ci-Ci ₂ )-alkanoyl-N-(Ci-Ci ₀ )-alkylamino, (C ₃ -C ₈ )- cycloalkanoyl-N-(C i -C ₁₀ )-alkylamino, (C ₆ -Ci ₂ )-aroyl-N-(C i -C ₁₀ )-alkylamino, (C ₇ -C ₁₁ )-aralkanoyl-N- (Ci-Cio)-alkylamino, amino-(Ci-Cio)-alkyl, (Ci-C ₂₀ )-alkylmercapto, (Ci-C ₂₀ )-alkylsulfinyl, (Ci-C ₂₀ )- alkylsulfonyl, (C ₆ -Ci ₂ )-arylmercapto, (C ₆ -C _]2 )-arylsulfinyl, (C ₆ -C ₎₂ )-arylsulfonyl, (C ₇ -Ci ₆ )- aralkylmercapto, (C ₇ -Ci ₆ )-aralkylsulfinyl, (C ₇ -Ci ₆ )-aralkylsulfonyl, sulfamoyl, N-(Ci-Ci ₀ )- alkylsulfamoyl, N,N-di-(C,-Ci ₀ )-alkylsulfamoyl, (C ₃ -C ₈ )-cycloalkylsulfamoyl, N-(C ₆ -Ci ₂ )- arylsulfamoyl, N-(C ₇ -Ci ₆ )-aralkylsulfamoyl, N-(Ci-Ci ₀ )-alkyl-N-(C ₆ -Ci ₂ )-arylsulfamoyl, N-(C ₁ -Ci ₀ )- alkyl-N-(C ₇ -C ₁₆ )-aralkylsulfamoyl, (Ci-Cio)-alkylsulfonamido, (C ₇ -Ci ₆ )-aralkylsulfonamido, and N- ((Ci-Cio)-alkyl-(C ₇ -Ci ₆ )-aralkylsulfonamido; where an aryl radical may be substituted by 1 to 5 substituents selected from hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl, (C ₂ -Ci ₆ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C ₃ -C ₈ )-cycloalkoxy, (C ₆ -C, ₂ )-aryl, (C ₇ -C, ₆ )-aralkyl, (C ₂ -C _I6 )-alkenyl, (C ₂ -C ₁₂ )- alkynyl, (C,-C ₁₆ )-alkoxy, (Ci-C ₁₆ )-alkenyloxy, (C ₆ -C ₁₂ )-aryloxy, (C ₇ -Ci ₆ )-aralkyloxy, (Ci-C ₈ )- hydroxyalkyl, -O-[CH ₂ ] _x C _f H _(2f+1 . _g) F _g , -OCF ₂ Cl, and -OCF ₂ -CHFCl; x is 0 to 3; f is 1 to 8; and g is 0 or 1 to (2f+l); including the physiologically active salts, esters, and prodrugs derived therefrom.

In one particular embodiment therein, the isoquinoline-3-carboxamide is selected from a compound of the Formula Ib wherein

A is -CHR ⁵ where R ⁵ is selected hydrogen or methyl;

B is -CO ₂ H;

X is O;

Q is O;

R ⁴ is hydrogen, (C ₁ -C ₃ )-alkyl, or substituted (C _r C ₃ )-alkyl;

R ³ is hydrogen, chloro, or cyano; and

R ¹⁶ , R ¹⁷ , R ¹⁸ and R ¹⁹ are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, aryl, heteroaryl, substituted heteroaryl, -OR ⁷⁰ , -SR ⁷⁰ , -SOR ⁷⁰ , and -SO ₂ R ⁷⁰ wherein R ⁷⁰ is selected from the group consisting of alkyl, substituted alkyl, cyclcoalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl, and substituted heteroaryl; including the physiologically active salts, esters, and prodrugs derived therefrom.

In other embodiments, isoquinoline-3-carboxamides for use in the present invention include those disclosed in International Publication No. WO 2004/108681 and as represented by Formula IV, IVA, IVB, IVC, IVD, VA, VB, VC and VD below.

As discussed, supra, compounds according the the present invention are in some embodiments heterocyclic carboxamides. In certain embodiments, heterocyclic carboxamides for use in the invention may be thienopyridine carboxamides. In particular embodiments, the thienopyridine carboxamide is selected from a thienopyridine-5 -carboxamide or a thienopyridine-6-carboxamide. In another embodiment, thienopyridine carboxamide compounds for use in the present invention are as disclosed in International Publication No. WO 2006/094292, represented by Formula II

(π) wherein

R ³⁰ is selected from the group consisting of hydrogen, (Ci-C ₆ )-alkyl, (C ₃ -C ₇ )-cycloalkyl, aryl, or a substituent of the α-carbon atom of an α-amino acid, wherein the amino acid is a natural L-amino acid or its D-isomer;

B is -CO ₂ H or a CO ₂ -G carboxyl radical, where G is a radical of an alcohol G-OH in which G is selected from the group consisting of (Ci-C ₂ o)-alkyl radical, (C ₃ -C ₈ ) cycloalkyl radical, (C ₂ -C ₂₀ )- alkenyl radical, (C ₃ -C ₈ )-cycloalkenyl radical, retinyl radical, (C ₂ -C ₂₀ )-alkynyl radical, (C ₄ -C ₂₀ )- alkenynyl radical; R ³¹ is selected from the group consisting of hydrogen, (Ci-Cio)-alkyl, (C ₂ -Ci ₀ )-alkenyl, (C ₂ -Ci ₀ )- alkynyl, wherein alkenyl or alkynyl contains one or two C-C multiple bonds; unsubstituted fluoroalkyl radical of the formula -[CH ₂ ] _x -CiH _(2f+1 . _g) -F _g , aryl, heteroaryl, and (C ₇ -Ci i)-aralkyl; one of D or M is -S-, and the other is =C(R ³⁴ )-; R ³² , R ³³ , and R ³⁴ are identical or different and are selected from the group consisting of hydrogen, hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl; (C _r C ₂₀ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C ₃ -

C ₈ )-cycloalkoxy, (C ₆ -Ci ₂ )-aryl, (C ₇ -Ci ₆ )-aralkyl, (C ₇ -Ci ₆ )-aralkenyl, (C ₇ -Ci ₆ )-aralkynyl, (C ₂ -C ₂₀ )- alkenyl, (C ₂ -C ₂₀ )-alkynyl, (Ci-C ₂₀ )-alkoxy, (C ₂ -C ₂₀ )-alkenyloxy, (C ₂ -C ₂₀ )-alkynyloxy, retinyloxy, (C ₆ - C ₁₂ )-aryloxy, (C ₇ -C, ₆ )-aralkyloxy, (C,-C, ₆ )-hydroxyalkyl, -O-[CH ₂ ] _x CfH _(2f+ i _-g) F _g , -OCF ₂ Cl, -OCF ₂ - CHFCl, (C _r C ₂₀ )-alkylcarbonyl, (C ₃ -C ₈ )-cycloalkylcarbonyl, (C ₆ -C, ₂ )-arylcarbonyl, (C ₇ -Ci ₆ )- aralkylcarbonyl, cinnamoyl, (C ₂ -C ₂₀ )-alkenylcarbonyl, (C ₂ -C ₂₀ )-alkynylcarbonyl, (Ci-C ₂₀ )- alkoxycarbonyl, (C ₆ -C] ₂ )-aryloxycarbonyl, (C ₇ -Ci ₆ )-aralkoxycarbonyl, (C ₃ -C ₈ )-cycloalkoxycarbonyl, (C ₂ -C ₂₀ )-alkenyloxycarbonyl, retinyloxycarbonyl, (C ₂ -C ₂₀ )-alkynyloxycarbonyl, (Ci-C] ₂ )- alkylcarbonyloxy, (C ₃ -C ₈ )-cycloalkylcarbonyloxy, (C ₆ -Ci ₂ )-arylcarbonyloxy, (C ₇ -Ci ₆ )- aralkylcarbonyloxy, cinnamoyloxy, (C ₂ -Ci ₂ )-alkenylcarbonyloxy, (C ₂ -Ci ₂ )-alkynylcarbonyloxy, (Ci- Ci ₂ )-alkoxycarbonyloxy, (C ₆ -Ci ₂ )-aryloxycarbonyloxy, (C ₇ -Ci ₆ )-aralkyloxycarbonyloxy, (C ₃ -C ₈ )- cycloalkoxycarbonyloxy, (C ₂ -Ci ₂ )-alkenyloxycarbonyloxy, (C ₂ -Ci ₂ )-alkynyloxycarbonyloxy, carbamoyl, N-(Ci-Ci ₂ )-alkylcarbamoyl, N,N-di-(Ci-Ci ₂ )-alkylcarbamoyl, N-(C ₃ -C ₈ )- cycloalkylcarbamoyl, N,N-dicyclo-(C ₃ -C ₈ )-alkylcarbamoyl, N-(Ci-Cio)-alkyl-N-(C ₃ -C ₈ )- cycloalkylcarbamoyl, N-((C ₃ -C ₈ )-cycloalkyl-(Ci-C ₆ )-alkyl)-carbamoyl, N-(+)- dehydroabietylcarbamoyl, N-(Ci-C ₆ )-alkyl-N-(+)-dehydroabietylcarbamoyl, N-(C ₆ -Ci ₂ )- arylcarbamoyl, N-(C ₇ -Ci ₆ )-aralkylcarbamoyl, N-(C _I -Ci ₀ )-alkyl-N-(C ₆ -Ci ₆ )-arylcarbamoyl, N-(C _r Cio)-alkyl-N-(C ₇ -Ci ₆ )-aralkylcarbamoyl, carbamoyloxy, N-(Ci-Ci ₂ )-alkylcarbamoyloxy, N,N-di-(Ci- Ci ₂ )-alkylcarbamoyloxy, N-(C ₃ -C ₈ )-cycloalkylcarbamoyloxy, N-(C ₆ -C i ₂ )-arylcarbamoyloxy, N-(C ₇ - c, ₆ )-aralkylcarbamoyloxy, N-(C ₁ -Cio)-alkyl-N-(C ₆ -C, ₂ )-arylcarbamoyloxy, N-(Ci-C,o)-alkyl-N-(C ₇ - Ci ₆ )-aralkylcarbamoyloxy, N-((C, -Ci ₀ )-alkyl)-carbamoyloxy, N-(C, -C, ₀ )-alkyl-N-((C ₇ -C _I6 )- aralkyloxy-(Ci-Ci ₀ )-alkyl)-carbamoyloxyamino, (Ci-Ci ₂ )-alkylamino, di-(Ci-Ci ₂ )-alkylamino, (C ₃ - C ₈ )-cycloalkylamino, (C ₃ -Ci ₂ )-alkenylamino, (C ₃ -Ci ₂ )-alkynylamino, N-(C ₆ -Ci ₂ )-arylamino, N-(C ₇ - Cii)-aralkylamino, N-alkyl-aralkylamino, N-alkyl-arylamino, (Ci-Ci ₂ )-alkoxyamino, (Ci-Ci ₂ )-alkoxy- N-(C ₁ -Cio)-alkylamino, (Ci-Ci ₂ )-alkanoylamino, (C ₃ -C ₈ )-cycloalkanoylamino, (C ₆ -Ci ₂ )-aroylamino, (C ₇ -Ci ₆ )-aralkanoylamino, (C ₁ -Ci ₂ )-alkanoyl-N-(Ci-Cio)-alkylamino, (C ₃ -C ₈ )-cycloalkanoyl-N-(Ci- Cio)-alkylamino, (C ₆ -Ci ₂ )-aroyl-N-(Ci-C ₁₀ )-alkylamino, (C ₇ -C ₁₁ )-aralkanoyl-N-(Ci-Cio)-alkylamino, amino-(Ci-Cio)-alkyl, (Ci-C ₂ o)-alkylmercapto, (C ₁ -C ₂ o)-alkylsulfϊnyl, (Ci-C ₂ o)-alkylsulfonyl, (C ₆ -

Ci ₂ )-arylmercapto, (C ₆ -Ci ₂ )-arylsulfinyl, (C ₆ -C ₁₂ )-arylsulfonyl, (C ₇ -Ci ₆ )-aralkylmercapto, (C ₇ -Ci ₆ )- aralkylsulfmyl, (C ₇ -C, ₆ )-aralkylsulfonyl, sulfamoyl, N-(C,-Ci ₀ )-alkylsulfamoyl, N ₉ N-(U-(C ₁ -Ci ₀ )- alkylsulfamoyl, (C ₃ -C ₈ )-cycloalkylsulfamoyl, N-(C ₆ -Ci ₂ )-arylsulfamoyl, N-(C ₇ -Ci ₆ )-aralkylsulfamoyl, N-(C,-C,o)-alkyl-N-(C ₆ -C ₁₂ )-arylsulfamoyl, N-(C,-C,o)-alkyl-N-(C ₇ -C ₁₆ )-aralkylsulfamoyl, (Ci-Co)- alkylsulfonamido, (C ₇ -Ci ₆ )-aralkylsulfonamido, and N-((Ci-Ci ₀ )-alkyl-(C ₇ -Ci ₆ )-aralkylsulfonamido; where an aryl radical may be substituted by 1 to 5 substituents selected from hydroxyl, halogen, cyano, trifluoromethyl, nitro, carboxyl, (C ₂ -C ₎₆ )-alkyl, (C ₃ -C ₈ )-cycloalkyl, (C ₃ -C ₈ )-cycloalkoxy, (C ₆ - C ₁₂ )-aryl, (C ₇ -Ci ₆ )-aralkyl, (C ₂ -Ci ₆ )-alkenyl, (C ₂ -C ₁₂ )-alkynyl, (C,-C, ₆ )-alkoxy, (C _r C ₁₆ )-alkenyloxy, (C ₆ -C ₁₂ )-aryloxy, (C ₇ -C, ₆ )-aralkyloxy, (C,-C ₈ )-hydroxyalkyl, -O-[CH ₂ ] _x C _f H _(2f+1 . _g) F _g , -OCF ₂ Cl, and - OCF ₂ -CHFCl; x is 0 to 3; f is 1 to 8; and g is 0 or 1 to (2f+l); including the physiologically active salts, esters, and prodrugs derived therefrom.

In certain embodiments, the compound is a compound of Formula II wherein

B is CO ₂ H;

R ³⁰ and R ³¹ are hydrogen;

R ³² is selected from hydrogen, halo, aryl, substituted aryl, aryloxy, and substituted aryloxy;

R ³³ is selected from hydrogen, halo, cyano, alkyl, alkynyl, and heteroaryl; one of D or M is -S-, and the other is =C(R ³⁴ )-; and

R ³⁴ is hydrogen, aryl, or substituted aryl; including the physiologically active salts, esters, and prodrugs derived therefrom.

As discussed herein, cyclic carboxamides are particularly suited for use in the present invention. However, use of other compounds that inhibit HIF prolyl hydroxylase activity is specifically contemplated. Such compounds are have been identified and are well-known in the art. For example, compounds according to the invention can include phenanthrolines and iron chelators, etc. In one embodiment, the compound for use in the present invention is an iron chelator, e.g., a hydroxamate. In particular embodiments, hydroxamates for use in the methods of the invention are selected from a compound of the formula (EI) or pharmaceutically acceptable salts thereof, wherein: a is an integer from 1 to 4; b is an integer from 0 to 4; c is an integer from 0 to 4;

Z is selected from the group consisting Of (C ₃ -Ci ₀ ) cycloalkyl, (C ₃ -Ci ₀ ) cycloalkyl independently substituted with one or more Y ¹ , 3-10 membered heterocycloalkyl and 3-10 membered heterocycloalkyl independently substituted with one or more Y ¹ ; (C ₅ -C ₂₀ ) aryl, (C ₅ -C ₂₀ ) aryl independently substituted with one or more Y ¹ , 5-20 membered heteroaryl and 5-20 membered heteroaryl independently substituted with one or more Y ¹ ;

Ar ¹ is selected from the group consisting of (C ₅ -C ₂₀ ) aryl, (C ₅ -C ₂₀ ) aryl independently substituted with one or more Y ² , 5-20 membered heteroaryl and 5-20 membered heteroaryl independently substituted with one or more Y ² ; each Y ¹ is independently selected from the group consisting of a lipophilic functional group, (C ₅ - C ₂₀ ) aryl, (C ₆ -C ₂₆ ) alkaryl, 5-20 membered heteroaryl and 6-26 membered alk-heteroaryl; each Y ² is independently selected from the group consisting of -R', -OR', -OR", -SR', -SR", - NR'R', -NO ₂ , -CN, -halogen, -trihalomethyl, trihalomethoxy, -C(O)R', -C(O)OR', - C(O)NR ¹ R', -C(O)NR ¹ OR', -C(NR ¹ R)=NOR', -NR'-C(O)R', -SO ₂ R', -SO ₂ R", -NR-SO ₂ - R', -Mf-C(O)-NR ¹ R ¹ , tetrazol-5-yl, -NR'-C(O)-OR', -C(NR'R')=NR', -S(O)-R', -S(O)-R", and -NR'-C(S)-NR'R'; and each R' is independently selected from the group consisting of -H, (Ci-C ₈ ) alkyl, (C ₂ -C ₈ ) alkenyl, and (C ₂ -C ₈ ) alkynyl; and each R" is independently selected from the group consisting of (C ₅ -C ₂₀ ) aryl and (C ₅ -C ₂₀ ) aryl independently substituted with one or more -OR', -SR', -NR'R', -NO ₂ , -CN, halogen or trihalomethyl groups, or wherein c is 0 and Ar ¹ is an N' substituted urea-aryl, the compound has the structural formula (ma):

or pharmaceutically acceptable salts thereof, wherein: a, b, and Z are as defined above; and R ³⁵ and R ³⁶ are each independently selected from the group consisting of hydrogen, (Ci-Cg) alkyl, (C ₂ -C ₈ ) alkenyl, (C ₂ -C ₈ ) alkynyl, (C ₃ -Ci ₀ ) cycloalkyl, (C ₅ -C ₂₀ ) aryl, (C ₅ -C ₂₀ ) substituted aryl, (C ₆ -C ₂₆ ) alkaryl, (C ₆ -C ₂₆ ) substituted alkaryl, 5-20 membered heteroaryl, 5-20 membered substituted heteroaryl, 6-26 membered alk-heteroaryl, and 6-26 membered substituted alk-heteroaryl; and

R ³⁷ is independently selected from the group consisting of hydrogen, (Ci-C ₈ ) alkyl, (C ₂ -C ₈ ) alkenyl, and (C ₂ -C ₈ ) alkynyl.

As discussed previously, compounds according to the present invention include isoquinoloine caarboxamides. In certain embodiments, compounds for use in the invention are isoquinoline-3- carboxamides. In certain embodiments, the compounds used in the present invention are as disclosed in International Publication No. WO 2004/108681, represented by formula (IV):

wherein: q is zero or one; p is zero or one;

R ^a is -COOH or -WR ⁸ ; provided that when R ^a is -COOH then p is zero and when R ^a is -WR ⁸ then p is one;

W is selected from the group consisting of oxygen, -S(O) _n - and -NR ⁹ - where n is zero, one or two, R ⁹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic and R ⁸ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, or when W is -NR ⁹ - then R ⁸ and R ⁹ , together with the nitrogen atom to which they are bound, can be joined to form a heterocyclic or a substituted heterocyclic group, provided that when W is -S(O) _n - and n is one or two, then R ⁸ is not hydrogen;

R ¹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, amino, substituted amino, aminoacyl, aryl, substituted aryl, halo, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl or, when X is -NR ⁷ -, then R ⁷ and R ⁸ , together with the nitrogen atom to which they are bound, can be joined to form a heterocyclic or substituted heterocyclic group;

R ² and R ³ are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano, -S(O) _n -N(R ⁶ )-R ⁶ where n is 0, 1 , or 2, -NR ⁶ C(O)NR ⁶ R ⁶ , -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, each R ⁶ is independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, cycloalkyl, substituted cycloalkyl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic provided that when X is -SO- or -SO ₂ -, then R ₆ is not hydrogen, and R ⁷ is selected from the group consisting of hydrogen, alkyl, aryl, or R ² , R ³ together with the carbon atom pendent thereto, form an aryl substituted aryl, heteroaryl, or substituted heteroaryl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl or, when X is -NR ⁷ -, then R ⁷ and R ⁸ , together with the nitrogen atom to which they are bound, can be joined to form a heterocyclic or substituted heterocyclic group;

R is selected from the group consisting of hydrogen, deuterium and methyl;

R ¹ is selected from the group consisting of hydrogen, deuterium, alkyl and substituted alkyl; alternatively, R and R' and the carbon pendent thereto can be joined to form cycloalkyl, substituted cycloalkyl, heterocyclic or substituted heterocyclic group; R" is selected from the group consisting of hydrogen and alkyl or R" together with R' and the nitrogen pendent thereto can be joined to form a heterocyclic or substituted heterocyclic group;

R'" is selected from the group consisting of hydroxy, alkoxy, substituted alkoxy, acyloxy, cycloalkoxy, substituted cycloalkoxy, aryloxy, substituted aryloxy, heteroaryloxy, substituted heteroaryloxy, aryl, -S(O) _n -R ¹⁰ wherein R ¹⁰ is selected from the group consisting of alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, heteroaryl and substituted heteroaryl and n is zero, one or two; and pharmaceutically acceptable salts, esters and prodrugs thereof. In an alternative embodiment, the compounds of formula (IV) are represented by formula (FVA):

wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R, R', R", R'" and q are as defined above; and pharmaceutically acceptable salts, esters, prodrugs thereof.

In an another alternative embodiment, the compounds of formula (FV) are represented by the formula (IVB):

wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R", R'", WR ⁸ and q are as defined above; and pharmaceutically acceptable salts, esters, prodrugs thereof.

In an another alternative embodiment, the invention is directed to compounds represented by the formula (IVC):

wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R, R', R", R'", WR ⁸ and q are as defined above; and pharmaceutically acceptable salts, esters, prodrugs thereof.

In yet another alternative embodiment, the invention is directed to compounds represented by the formula (IVD):

wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R, R', R", R'" and q are as defined above; and pharmaceutically acceptable salts, esters, prodrugs thereof. In other embodiments, the invention is directed to isoquinoline carboxamide compounds represented by the formulae (VA), (VB), (VC), (V), wherein said formulae are defined below.

Formula VA:

wherein: q is zero or one;

R ¹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, halo, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl;

R ² and R ³ are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano, -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl;

R is selected from the group consisting of hydrogen and methyl;

R' is selected from the group consisting of alkyl and substituted alkyl; or R and R' may be joined to form a cycloalkyl, substituted cycloalkyl, heterocyclic or substituted heterocyclic; R" is selected from the group consisting of hydrogen and alkyl or R" together with R' and the nitrogen pendent thereto forms a heterocyclic or substituted heterocyclic group; or pharmaceutically acceptable salts and/or prodrugs thereof. Formula VB:

wherein: q is zero or one; W is selected from the group consisting of oxygen, -S(O) _n - and -NR ⁹ - where n is zero, one or two,

R ⁹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, acyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁸ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic;

R" is selected from hydrogen and alkyl;

R ¹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, halo, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl; R ² and R ³ are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano, -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is * hydrogen, alkyl or aryl; or pharmaceutically acceptable salts and/or prodrugs thereof. Formula VC:

wherein: q is zero or one; R ¹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, halo, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl, or aryl;

R ² and R ³ are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano, -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl, or aryl;

R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl, or aryl;

R is selected from the group consisting of hydrogen and methyl;

R' is selected from the group consisting of alkyl and substituted alkyl; or R and R' can be joined to form cycloalkyl, substituted cycloalkyl, heterocyclic or substituted heterocyclic,

R" is selected from the group consisting of hydrogen and alkyl or R" together with R' and the nitrogen pendent thereto forms a heterocyclic or substituted heterocyclic group;

W is selected from the group consisting of oxygen, -S(O) _n - and -NR ⁹ - where n is zero, one or two, R ⁹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁸ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic; or pharmaceutically acceptable salts and/or prodrugs thereof.

Formula VD:

wherein: q is zero or one;

R" is selected from hydrogen and alkyl;

R ¹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, halo, heteroaryl, substituted heteroaryl, heterocyclic, substituted heterocyclic, and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl;

R ² and R ³ are independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, halo, hydroxy, cyano, -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl; R ⁴ and R ⁵ are independently selected from the group consisting of hydrogen, halo, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, substituted aryl, heteroaryl, substituted heteroaryl and -XR ⁶ where X is oxygen, -S(O) _n - or -NR ⁷ - where n is zero, one or two, R ⁶ is selected from the group consisting of alkyl, substituted alkyl, aryl, substituted aryl, heteroaryl, substituted heteroaryl, heterocyclic and substituted heterocyclic, and R ⁷ is hydrogen, alkyl or aryl; or pharmaceutically acceptable salts and/or prodrugs thereof.

In compounds of formulae (IV), (IVA), (IVB), (IVC), and (IVD), preferably R ¹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, halo, alkoxy, aryloxy, substituted aryloxy, substituted aryl, alkylthio, aminoacyl, aryl, substituted amino, heteroaryl, heteroaryloxy, -S(O) _n -aryl, -S(O) _n -substituted aryl, -S(O) _n -heteroaryl, and -S(O) _n -substituted heteroaryl, where n is zero, one or two. More preferably, R ¹ is selected from the group consisting of: (3-methoxyphenyl)sulfanyl; (4- chlorophenyl)sulfanyl; (4-methylphenyl)sulfanyl; 2-fluorophenoxy; 2-methoxyphenoxy; (2- methoxyphenyl)sulfanyl 3-fluorophenoxy; 3-methoxyphenoxy; 4-(methylcarbonylamino)phenoxy; A- (methylsulfonamido)phenoxy; 4-fluorophenoxy; 4-methoxyphenoxy; 4-methoxyphenylsulfanyl; A- methylphenyl; bromo; cMoro; dimethylaminomethyl; ethoxy; ethylsulfanyl; hydrogen; isopropyl; methoxy; methoxymethyl; methyl; N,N-dimethylaminocarbonyl; naphth-2-yloxy; naphthylsulfanyl; phenoxy; phenyl; phenylamino; phenylsulfinyl; phenylsulfanyl; pyridin-2-yloxy; pyridin-2-yl; and pyridin-2-ylsulfanyl.

In compounds of formulae (IV), (IVA), (IVB), (IVC), and (IVD), R ² is preferably selected from the group consisting of substituted amino, aryloxy, substituted aryloxy, alkoxy, substituted alkoxy, halo, hydrogen, alkyl, substituted alkyl, aryl, -S(O) _n -aryl, -S(O) _n -substituted aryl, -S(O) _n -cycloalkyl, where n is zero, one or two, aminocarbonylamino, heteroaryloxy, and cycloalkyloxy. More preferably, R ² is selected from the group consisting of: (4-methoxy)phenylsulfonylamino; 2,6-dimethylphenoxy; 3,4-difluorophenoxy; 3,5- difluorophenoxy; 3-chloro-4-fluorophenoxy; 3-methoxy-4-fluorophenoxy; 3-methoxy-5-fluorophenoxy;

4-(methylsulfonamido)phenoxy; 4-(phenylsulfonamido)phenoxy; 4-CF ₃ -O-phenoxy; 4-CF ₃ -phenoxy; A- chlorophenoxy; 4-fluorophenoxy; 4-(4-fluorophenoxy)phenoxy; 4-methoxyphenoxy; 4-nitrophenoxy; benzyloxy; bromo; butoxy; CF ₃ ; chloro; cyclohexyloxy; cyclohexylsulfanyl; cyclohexylsulfonyl; fluoro; hydrogen; iodo; isopropoxy; methyl; phenoxy; phenyl; phenylsulfanyl; phenylsulfinyl; phenylsulfonyl; phenylurea; pyridin-1-ylsulfanyl; pyridin-3-yloxy; and pyridin-4-ylsulfanyl.

In compounds of formulae (FV), (FVA), (IVB), (IVC), and (FVD), R ³ is preferably selected from the group consisting of: substituted aryloxy, substituted alkoxy, alkoxy, substituted alkyl, alkyl, amino, cycloalkyloxy, hydrogen, halo, aryl, -S(O) _n -aryl, -S(O) _n -substituted aryl, -S(O) _n -heteroaryl, and -S(O) _n -substituted heteroaryl, where n is zero, one or two, aminocarbonylamino, and heteroaryloxy.

More preferably, R ³ is selected from the group consisting of: amino; (4- methyl)phenylsulfonylaminophenoxy; 3,4-difluorophenoxy; 3,5-difluorophenoxy; 3-fluoro-5-methoxy- phenoxy; 3-chloro-4-fluorophenoxy; 4-CF ₃ -O-phenoxy; 4-CF ₃ -phenoxy; 4-chlorophenoxy; A- fluorophenoxy; 4-(4-fluorophenoxy)phenoxy; 4-methoxyphenoxy; benzyloxy; bromo; butoxy; CF ₃ ; chloro; cyclohexyloxy; hydrogen; iodo; isopropoxy; phenoxy; phenyl; phenylsulfanyl; phenylsulfonyl; phenylsulfinyl; phenylurea; pyridin-1-ylsulfanyl; pyridin-3-yloxy; and pyridin-4-ylsulfanyl.

Alternatively, R ² and R ³ , combined with the carbon atoms pendent thereto, are joined to form an aryl group. Preferably, the aryl group is phenyl. In compounds of formulae (IV), (IVA), (IVB), (IVC), and (IVD), R ⁴ is preferably selected from the group consisting of: substituted arylthio, halo, hydrogen, substituted alkyl and aryl.

More preferably, R ⁴ is selected from the group consisting of: 4-chlorophenyl sulfanyl; chloro; hydrogen; methoxymethyl; and phenyl.

In compounds of formulae (IV), (IVA), (IVB), (IVC), and (IVD), R ⁵ is preferably hydrogen or aryl. More preferably R ⁵ is hydrogen or phenyl.

In compounds of formulae (IV), (FVA) and (FVC), R is preferably selected from the group consisting of hydrogen, deuterium, aryl and alkyl. More preferably R is selected from the group consisting of phenyl, hydrogen, deuterium and methyl.

In compounds of formulae (FV), (FVA) and (FVC), R' is selected from the group consisting of preferably hydrogen, deuterium, alkyl, substituted alkyl, and substituted amino. More preferably, R' is selected from the group consisting of: 4-aminobutyl; 4-hydroxybenzyl; benzyl; carboxylmethyl; deuterium; hydroxymethyl; imidazol-4-ylmethyl; isopropyl; methyl; and propyl.

Alternatively, R, R' and the carbon atom pendent thereto join to form a cycloalkyl and more preferably cyclopropyl.

In compounds of formulae (FV), (FVA) and (FVC), R" is preferably hydrogen, alkyl or substituted alkyl. More preferably, R" is hydrogen, methyl or carboxylmethyl (-CH ₂ C(O)OH). Alternatively, R', R" and the carbon atom and nitrogen atom respectively pendent thereto join to form a heterocyclic group and more preferably pyrrolidinyl.

In compounds of formulae (FV), (FVA), (FVB), (FVC) and (FVD), preferably R'" is selected from the group consisting of hydrogen, hydroxy, alkoxy, substituted alkoxy, cycloalkoxy, substituted cycloalkoxy, thiol, acyloxy and aryl. Preferably, R'" is selected from the group consisting of: hydroxy; benzyloxy; ethoxy; thiol; methoxy; methylcarbonyloxy; and phenyl.

In compounds of formulae (FV), (FVB) and (FVC), WR ⁸ is preferably selected from the group consisting of amino, substituted amino, aminoacyl, hydroxy, and alkoxy. More preferably, WR ⁸ is selected from the group consisting of: amino; dimethylamino; hydroxy; methoxy; and methylcarbonylamino. As discussed, supra, compounds according the the present invention are in some embodiments heterocyclic carboxamides; in particular, pyrrolopyridazine carboxamides. In certain embodiments, compounds for use in the invention are pyrrolopyridazine-3 -carboxamides. In one embodiment, the compound is a compound of formula VII:

wherein

R ⁵² is selected from the group consisting of hydrogen, alkyl, and substituted alkyl;

R ⁵³ is selected from the group consisting of hydrogen, deuterium, alkyl, and substituted alkyl;

R ⁵⁴ is selected from the group consisting of hydrogen, deuterium, alkyl, and substituted alkyl; R ⁵⁶ , R ⁵⁷ and R ⁵⁸ independently are selected from the group consisting of hydrogen, hydroxy, cyano, halo, nitro, acyl, amino, substituted amino, acylamino, sulfonyl, substituted sulfonyl, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, alkoxy, substituted alkoxy, cycloalkyl, substituted cycloalkyl, cycloalkyloxy, substituted cycloalkyloxy, heterocyclyloxy, substituted heterocyclyloxy, carboxyl, carboxyl ester, carboxyl amide, oxycarbonylamino, aminocarbonyloxy, aminocarbonylamino, aryl, substituted aryl, aryloxy, substituted aryloxy, heteroaryloxy, substituted heteroaryloxy, alkylthio, substituted alkylthio, cycloalkylthio, substituted cycloalkylthio, arylthio, substituted arylthio, heteroarylthio, substituted heteroarylthio, hetereocyclicthio, substituted heterocyclicthio, heteroaryl, and substituted heteroaryl; or wherein R ⁵⁶ and R ⁵⁷ , or R ⁵⁷ and R ⁵⁸ , together with the carbons to which they are attached, form a carbocyclic 5- or 6-membered aromatic ring, optionally substituted by one or two hydrogen, halogen, alkyl, substituted alkyl, alkoxy, substituted alkoxy, aryl, or substituted aryl;

R ⁵⁹ is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, heterocyclic, substituted heterocyclic, aryl, substituted aryl, heteroaryl, and substituted heteroaryl;

R ⁶⁰ is -NR ⁶¹ R ⁶² or -OR ⁶³ ;

R ⁶¹ and R ⁶² independently are selected from the group consisting hydrogen, alkyl, alkylene- cycloalkyl, C ₃ -C ₈ heterocyclic, aryl, and -C(O)(Ci-C ₄ alkyl); or R ⁶¹ and R ⁶² taken together with the nitrogen to which they are attached form a 5- or 6- membered heterocyclic or substituted heterocyclic; and R ⁶³ is selected from the group consisting of hydrogen and alkyl which is unsubstituted or substituted with one or more substituents independently selected from the group consisting of cycloalkyl, heterocyclic, aryl, and heteroaryl; or a pharmaceutically acceptable salt, single stereoisomer, mixture of stereoisomers, ester, tautomer or prodrug thereof.

In certain embodiments, the invention relates to compounds of Formula VII wherein R ⁵² and R ⁵³ are hydrogen;

R ⁵⁴ is selected from the group consisting of hydrogen and methyl;

R ⁵⁶ , R ⁵⁷ , and R ⁵⁸ independently are selected from the group consisting of hydrogen, halo, and aryl;

R ⁵⁹ is selected from the group consisting of hydrogen, alkyl, -CH ₂ -aryl, -CH ₂ -substituted aryl, or -

CH ₂ -heteroaryl; and R ⁶⁰ is -OR ⁶³ ; wherein R ⁶³ is hydrogen or alkyl.

In certain embodiments, the invention relates to compounds of Formula VII wherein

R ⁵² , R ⁵³ , and R ⁵⁴ are hydrogen;

R ⁵⁶ and R ⁵⁸ independently are selected from the group consisting of hydrogen and halo;

R ⁵⁷ is selected from the group consisting of hydrogen, halo, and aryl;

R ⁵⁹ is selected from the group consisting of hydrogen, alkyl, -CH ₂ -aryl, -CH ₂ -substituted aryl, or ■ CH ₂ -heteroaryl; and

R ⁶⁰ is -OR ⁶³ ; wherein R ⁶³ is hydrogen or alkyl.

The terms "hydroxy" or "hydroxyl" refer to the group -OH.

The term "halo" or "halogen" refers to fluoro, chloro, bromo, and iodo.

The term "cyano" refers to the group -CN.

The term "nitro" refers to the group -NO ₂ .

The term "carboxyl" refers to -COOH or salts thereof.

The term "alkyl" refers to saturated monovalent hydrocarbyl groups having from 1 to 10 carbon atoms; more particularly, from 1 to 5 carbon atoms; and, even more particularly, 1 to 3 carbon atoms. This term is exemplified by groups such as methyl, ethyl, n-propyl, iso-propyl, n-butyl, /-butyl, n-pentyl, and the like. The term "cycloalkyl" refers to a saturated or an unsaturated, but nonaromatic, cyclic alkyl groups of from 3 to 10, 3 to 8, or 3 to 6 carbon atoms having single or multiple cyclic rings including, by way of example, adamantyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclooctyl, cyclohexenyl, and the like.

The term "cycloalkoxy" refers to an -O-cycloalkyl group.

The term "aryl" refers to a monovalent aromatic carbocyclic group of from 6 to 14 carbon atoms having a single ring (e.g., phenyl) or multiple condensed rings (e.g., naphthyl or anthryl), which condensed rings may or may not be aromatic (e.g., 2-benzoxazolinone, 2H-l,4-benzoxazin-3(4H)-one-7-yl, and the like) provided that the point of attachment is the aryl group. Preferred aryls include phenyl and naphthyl.

The terms "heterocyclic" or "heterocyclyl" refer to a saturated or unsaturated ring system having a single ring or multiple condensed rings, from 1 to 10 carbon atoms, and from 1 to 4 hetero atoms selected from the group consisting of nitrogen, sulfur, or oxygen within the ring.

The term "heteroaryl" refers to an aromatic heterocyclic group of from 1 to 15 carbon atoms, preferably from 1 to 10 carbon atoms, and 1 to 4 heteroatoms within the ring selected from the group consisting of oxygen, nitrogen, and sulfur. Such heteroaryl groups can have a single ring (e.g., pyridinyl, furyl, or thienyl) or multiple condensed rings (e.g., indolizinyl or benzothienyl), which condensed rings may or may not be aromatic provided the point of attachment is through a ring containing the heteroatom and that ring is aromatic. The nitrogen can optionally be oxidized to provide for the N-oxide, and/or the sulfur ring atoms can optionally be oxidized to provide for the sulfoxide and sulfone derivatives.

Examples of heterocycles and heteroaryls include, but are not limited to, azetidine, pyrrole, imidazole, pyrazole, pyridine, pyrazine, furan, pyrimidine, pyridazine, indolizine, isoindole, indole, dihydroindole, indazole, purine, quinolizine, isoquinoline, quinoline, phthalazine, naphthylpyridine, quinoxaline, quinazoline, cinnoline, pteridine, carbazole, carboline, phenanthridine, acridine, phenanthroline, isothiazole, phenazine, isoxazole, phenoxazine, phenothiazine, imidazolidine, imidazoline, piperidine, piperazine, indoline, phthalimide, 1,2,3,4-tetrahydroisoquinoline, 4,5,6,7-tetrahydrobenzo[b]thiophene, thiazole, thiazolidine, thiophene, benzo[b]thiophene, morpholinyl, thiomorpholinyl (also referred to as thiamorpholinyl), piperidinyl, pyrrolidine, tetrahydrofuranyl, and the like.

The term "alkenyl" refers to a vinyl unsaturated monovalent hydrocarbyl group having from 2 to 6, preferably from 2 to 4, carbon atoms, and having at least 1, preferably from 1 to 2, sites of vinyl (>C=C<) unsaturation. Such groups are exemplified by vinyl (ethen-1-yl), allyl, but-3-enyl, and the like.

The term "alkynyl" refers to acetylinic unsaturated monovalent hydrocarbyl groups having from 2 to 6, preferably from 2 to 3, carbon atoms and having at least 1, preferably from 1 to 2, sites of acetylenic (-C ≡€-) unsaturation. This group is exemplified by ethyn-1-yl, propyn-1-yl, propyn-2-yl, and the like. The term "alkoxy" refers to the group "alkyl-O-," which includes, by way of example, methoxy, ethoxy, n-propoxy, iso-propoxy, n-butoxy, f-butoxy, sec-butoxy, n-pentoxy, and the like.

The term "alkenyloxy" refers to the group "alkenyl-O-."

The term "alkynyloxy" refers to the group "alkynyl-O-."

The term "aryloxy" refers to the group aryl-O- that includes, by way of example, phenoxy, naphthoxy, and the like.

The term "aralkyloxy" refers to the group aralkyl-O- that includes, by way of example, benzyloxy, and the like.

The term "carbonyl" refers to C=O.

The term "carbonyloxy" refers to -C(=0)O.

The terms "aminoacyl" or "amide", or the prefixes "carbamoyl" or "carboxamide," refer to the group -C(0)NR ^q R ^q where each R ^q is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, aryl, cycloalkyl, heteroaryl, and heterocyclic; or where each R ^q is joined to form together with the nitrogen atom a heterocyclic wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein.

The term "amino" refers to the group -NH ₂ .

The terms "thio" or "mercapto" refer to the group -SH.

The terms "alkylsulfanyl," "alkylthio," or "thioether" refer to the groups -S-alkyl where alkyl is as defined above.

The term "sulfinyl" refers to the group -S(O)-.

The term "sulfonyl" refers to the group -S(O) ₂ -.

The term "heterocyclyloxy" refers to the group -O-heterocyclic.

The term "cycloalkylene" refers to divalent cycloalkyl groups as defined above. The terms "cycloalkylthio" or "cycloalkylsulfanyl" refer to the groups -S-cycloalkyl where cycloalkyl is as defined herein.

The terms "arylthio" or "arylsulfanyl" refer to the group -S-aryl, where aryl is as defined herein. The terms "heteroarylthio" or "heteroarylsulfanyl" refer to the group -S-heteroaryl, where heteroaryl is as defined herein.

The terms "heterocyclicthio" or "heterocyclicsulfanyl" refer to the group -S-heterocyclic, where heterocyclic is as defined herein.

The term "alkyl alcohol" refers to the group "alkyl-OH". "Alkyl alcohol" is meant to include methanol, ethanol, 2-propanol, 2-butanol, butanol, etc.

The term "acyl" refers to the groups H-C(O)-, alkyl-C(O)-, alkenyl-C(O)-, alkynyl-C(O)-, cycloalkyl- C(O)-, aryl-C(O)-, heteroaryl-C(O)-, and heterocyclic-C(O)-, provided that a nitrogen atom of the heterocyclic is not bound to the -C(O)- group, wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein.

The term "acyloxy" refers to the groups alkyl-C(O)O-, alkenyl-C(O)O-, alkynyl-C(O)O-, aryl-C(O)O-, cycloalkyl-C(O)O-, heteroaryl-C(O)O-, and heterocyclic-C(O)O-, wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein.

The term "alkenyl" refers to a vinyl unsaturated monovalent hydrocarbyl group having from 2 to 6 carbon atoms, and preferably 2 to 4 carbon atoms, and having at least 1, and preferably from 1 to 2 sites of vinyl

(>C=C<) unsaturation. Such groups are exemplified by vinyl (ethen-1-yl), allyl, but-3-enyl and the like.

The term "alkynyl" refers to acetylinic unsaturated monovalent hydrocarbyl groups having from 2 to 6, preferably from 2 to 3, carbon atoms and having at least 1, preferably from 1 to 2, sites of acetylenic (-C ≡C-) unsaturation. This group is exemplified by ethyn-1-yl, propyn-1-yl, propyn-2-yl, and the like.

The term "acylamino" refers to the groups -NR'C(O)alkyl, -NR ¹ C(O)CyClOaIlCyI, -NR'C(O)alkenyl,

-NR ^t C(O)alkynyl,-NR'C(O)aryl, -NR ^t C(O)heteroaryl, and -NR'C(O)heterocyclic where R ¹ is hydrogen or alkyl, and wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic are defined herein.

The term "carbonyloxyamino" refers to the groups -NR ^u C(O)O-alkyl, -NR ^u C(O)O-alkenyl, -NR ^u C(0)0- alkynyl, -NR ^u C(O)O-cycloalkyl, -NR ^u C(O)O-aryl, -NR ^u C(O)O-heteroaryl, and -NR ^u C(O)O-heterocyclic, where R ^u is hydrogen or alkyl and wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein.

The term "oxycarbonylamino" refers to the groups -NR ^u C(O)O-alkyl, -NR ^u C(O)O-alkenyl, -NR ¹¹ C(O)O- alkynyl, -NR ^u C(O)O-cycloalkyl, -NR ^u C(O)O-aryl, -NR ^u C(O)O-heteroaryl, and -NR ^u C(O)O-heterocyclic, where R" is hydrogen or alkyl, and wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein. The term "oxythiocarbonylamino" refers to the groups -NR ^u C(S)O-alkyl,-NR ^u C(S)O-alkenyl, -NR ^u C(S)O-alkynyl, -NR ^u C(S)O-cycloalkyl, -NR ^u C(S)O-aryl, -NR ^u C(S)O-heteroaryl, and -NR ^U C(S)O- heterocyclic, where R ^u is hydrogen or alkyl, and wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic are as defined herein.

The term "aminocarbonyloxy" or the prefix "carbamoyloxy" refer to the groups -OC(O)NR ^V R ^V where each R ^v is independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, and heterocyclic; or where each R ^v is joined to form, together with the nitrogen atom, a heterocyclic, and wherein alkyl, alkenyl, alkynyl, cycloalkyl, aryl, heteroaryl, substituted heteroaryl, and heterocyclic are as defined herein.

The term "aminocarbonylamino" refers to the group -NR ^W C(O)N(R ^W ) ₂ where each R ^w is independently selected from the group consisting of hydrogen and alkyl.

The term "aminothiocarbonylamino" refers to the group -NR ^W C(S)N(R ^W ) ₂ where each R ^w is independently selected from the group consisting of hydrogen and alkyl.

The term "aryloxyaryl" refers to the group -aryl-O-aryl.

The term "carboxyl ester" refers to the groups -C(O)O-alkyl, -C(O)O-alkenyl, -C(O)O-alkynyl, -C(O)O- cycloalkyl, -C(O)O-aryl, -C(O)O-substituted aryl, -C(O)O-heteroaryl, -C(O)O-substituted heeteroaryl, -C(O)O-heterocyclic, and -C(O)O-substituted heterocyclic.

The term "cycloalkylene" refers to divalent cycloalkyl groups as defined above.

The term "heteroaryloxy" refers to the group -O-heteroaryl.

The term "sulfonyl" refers to the group -S(O) ₂ -, and may be included in the groups -S(O) ₂ H, -SO ₂ -alkyl,

-SO ₂ -alkenyl, -SO ₂ -alkynyl, -SO ₂ -cycloalkyl, -SO ₂ -cycloalkenyl, -SO ₂ -aryl, -SO ₂ -substituted aryl, -SO ₂ -heteroaryl, and -SO ₂ -heterocyclic, wherein alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, and heterocyclic are as defined herein.

The term "heterocyclyloxy" refers to the group -O-heterocyclic.

The terms "arylthio" or "arylsulfanyl" refer to the group -S-aryl.

The terms "heteroarylthio" or "heteroarylsulfanyl" refer to the group -S-heteroaryl.

The terms "heterocyclicthio" or "heterocyclicsulfanyl" refer to the group -S -heterocyclic. Conjugated terms refer to a linear arrangement of the separate substituents as each separate term is defined herein. For example, the term "aralkyl" refers to an aryl-alkyl group and includes, by way of example, benzyl; the term "aralkylcarbamoyl" refers to an aryl-alkyl-carbomoyl substituent wherein each term is as defined herein, etc.

It is understood that in all substituted and conjugated groups as defined herein, polymers arrived at by defining substituents with further substituents to themselves (e.g., aryl having a substituted aryl group as a substituent which is itself substituted with a substituted aryl group, etc.) are not intended for inclusion herein. Also not included are infinite numbers of substituents, whether the substituents are the same or different. In such cases, the maximum number of such substituents is three.

Similarly, it is understood that the above definitions are not intended to include impermissible substitution patterns (e.g., methyl substituted with 5 fluoro groups or a hydroxyl group alpha to ethenylic or acetylenic unsaturation). Such impermissible substitution patterns are well known to the skilled artisan.

The term "pharmaceutically acceptable salt" refers to pharmaceutically acceptable salts of a compound, which salts are derived from a variety of organic and inorganic counter ions well known in the art, and include, by way of example only, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, and the like; and, when the molecule contains a basic functionality, salts of organic or inorganic acids, such as hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, and the like.

The terms "stereoisomer" or "stereoisomers" refer to compounds that differ in the chirality of one or more stereocenters. Stereoisomers include enantiomers (compounds are non-superimposable mirror images) and diastereomers (compounds having more than one stereogenic center that are non-mirror images of each other and wherein one or more stereogenic center differs between the two stereoisomers). The compounds of the invention can be present as a mixture of stereoisomers or as a single stereoisomer.

The term "tautomer" refers to alternate forms of a compound that differ in the position of a proton, such as enol, keto, and imine enamine tautomers, or the tautomeric forms of heteroaryl groups contining a ring atom attached to both a ring NH moiety and a ring =N moiety such as pyrazoles, imidazoles, benzimidazoles, triazoles, and tetrazoles.

The term "prodrug," as used herein, refers to compounds that include chemical groups which, in vivo, can be converted into the carboxylate group and/or can be split off from the amide N-atom and/or can be split off from the R atom to provide for the active drug, a pharmaceutically acceptable salt thereof, or a biologically active metabolite thereof. Suitable groups are well known in the art and particularly include: for the carboxylic acid moiety, a prodrug selected from, e.g., esters including, but not limited to, those derived from alkyl alcohols, substituted alkyl alcohols, hydroxy substituted aryls and heteroaryls and the like; amides, particularly amides derived from amines of the Formula HNR ²⁰⁰ R ²¹⁰ where R ²⁰⁰ and R ²¹⁰ are independently hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, and the like; hydroxymethyl, aldehyde and derivatives thereof. The term "ester" refers to compounds that include the group -COOR where R is alkyl, substituted alkyl, alkoxy, or substituted alkoxy.

Methods for Identifying Compounds

Methods for identifying compounds of the invention are also provided. In certain aspects, a compound of the invention is one that stabilizes HIFα. The ability of a compound to stabilize or activate HIFa can be measured, for example, by direct measurement of HIFα in a sample, indirect measurement of HIFα, e.g., by measuring a decrease in HDFa associated with the von Hippel Lindau protein (see, e.g., International

Publication No. WO 2000/69908), or activation of HIF responsive target genes or reporter constructs (see, e.g., U.S. Patent No. 5,942,434). Measuring and comparing levels of HIF and/or HIF-responsive target proteins in the absence and presence of the compound will identify compounds that stabilize HIFα and/or activate HIF. Suitable compounds for use in the present methods may be identified and characterized using the assay described in International Publication No. WO 2005/118836, or in Example 10 of

International Publication No. WO 2003/049686, both of which are incorporated herein by reference in their entirety. Compounds identifiable by these assays are specifically envisaged for use in the present invention.

In other aspects, a compound of the invention is one that inhibits HIF hydroxylase activity. Assays for hydroxylase activity are standard in the art. Such assays can directly or indirectly measure hydroxylase activity. For example, an assay can measure hydroxylated residues, e.g., proline, asparagine, etc., present in the enzyme substrate, e.g., a target protein, a synthetic peptide mimetic, or a fragment thereof. (See, e.g., Palmerini et al. (1985) J Chromatogr 339:285-292.) A reduction in hydroxylated residue, e.g., proline or asparagine, in the presence of a compound is indicative of a compound that inhibits hydroxylase activity. Alternatively, assays can measure other products of the hydroxylation reaction, e.g., formation of succinate from 2-oxoglutarate. (See, e.g., Cunliffe et al. (1986) Biochem J 240:617- 619.) Kaule and Gunzler (1990; Anal Biochem 184:291-297) describe an exemplary procedure that measures production of succinate from 2-oxoglutarate.

Procedures such as those described above can be used to identify compounds that modulate HIF hydroxylase activity. Target protein may include HIFα or a fragment thereof, e.g., HIF(556-575). Enzyme may include, e.g., HIF prolyl hydroxylase (see, e.g., GenBank Accession No. AAG33965, etc.) or HIF asparaginyl hydroxylase (see, e.g., GenBank Accession No. AAL27308, etc.), obtained from any source. Enzyme may also be present in a crude cell lysate or in a partially purified form. For example, procedures that measure HIF hydroxylase activity are described in Ivan et al. (2001, Science 292:464-468; and 2002, Proc Natl Acad Sci USA 99:13459-13464) and Hirsila et al. (2003, J Biol Chem 278:30772-30780); additional methods are described in International Publication No. WO 03/049686. Measuring and comparing enzyme activity in the absence and presence of the compound will identify compounds that inhibit hydroxylation of HIFα.

Pharmaceutical Formulations and Routes of Administration

The compositions of the present invention can be delivered directly or in pharmaceutical compositions containing excipients, as is well known in the art. The present methods of treatment involve administration of an effective amount of a compound of the present invention to a subject in need, wherein the subject would benefit from increased neurogenesis, neural cell differentiation, dopaminergic differentiation, or neural cell proliferation.

An effective amount, e.g., dose, of compound or drug can readily be determined by routine experimentation, as can an effective and convenient route of administration and an appropriate formulation. Various formulations and drug delivery systems are available in the art. (See, e.g., Gennaro, ed. (2000) Remington's Pharmaceutical Sciences, supra; and Hardman, Limbird, and Gilman, eds. (2001)

The Pharmacological Basis of Therapeutics, supra.)

Suitable routes of administration may, for example, include oral, rectal, topical, nasal, pulmonary, ocular, intestinal, and parenteral administration. Primary routes for parenteral administration include intravenous, intramuscular, and subcutaneous administration. Secondary routes of administration include intraperitoneal, intra-arterial, intra-articular, intracardiac, intracisternal, intradermal, intralesional, intraocular, intrapleural, intrathecal, intrauterine, and intraventricular administration. The indication to be treated, along with the physical, chemical, and biological properties of the drug, dictate the type of formulation and the route of administration to be used, as well as whether local or systemic delivery would be preferred.

In preferred embodiments, the compounds of the present invention are administered orally. For example, in certain embodiments, the present invention provides for oral administration of [(4-Hydroxy-7- phenylsulfanyl-isoquinoline-3-carbonyl)-amino]-acetic acid (Compound A), [(4-Hydroxy-7-phenoxy- isoquinoline-3-carbonyl)-amino] -acetic acid (Compound B), {[l-Cyano-6-(2,6-dimethyl-phenoxy)-4- hydroxy-isoquinoline-3-carbonyl] -amino} -acetic acid (Compound C), or {[4-Hydroxy-2-oxo-l-(4- trifluoromethyl-benzyl)-l,2-dihydro-pyrrolo[l,2-b]pyridazine -3-carbonyl]-amino} -acetic acid.

Pharmaceutical dosage forms of a compound of the invention may be provided in an instant release, controlled release, sustained release, or target drug-delivery system. Commonly used dosage forms include, for example, solutions and suspensions, (micro-) emulsions, ointments, gels and patches, liposomes, tablets, dragees, soft or hard shell capsules, suppositories, ovules, implants, amorphous or crystalline powders, aerosols, and lyophilized formulations. Depending on route of administration used, special devices may be required for application or administration of the drug, such as, for example, syringes and needles, inhalers, pumps, injection pens, applicators, or special flasks. Pharmaceutical dosage forms are often composed of the drug, an excipient(s), and a container/closure system. One or multiple excipients, also referred to as inactive ingredients, can be added to a compound of the invention to improve or facilitate manufacturing, stability, administration, and safety of the drug, and can provide a means to achieve a desired drug release profile. Therefore, the type of excipient(s) to be added to the drug can depend on various factors, such as, for example, the physical and chemical properties of the drug, the route of administration, and the manufacturing procedure. Pharmaceutically acceptable excipients are available in the art, and include those listed in various pharmacopoeias. (See, e.g., USP, JP, EP, and BP,

FDA web page (www.fda.gov), Inactive Ingredient Guide 1996, and Handbook of Pharmaceutical Additives, ed. Ash; Synapse Information Resources, Inc. 2002.)

Pharmaceutical dosage forms of a compound of the present invention may be manufactured by any of the methods well-known in the art, such as, for example, by conventional mixing, sieving, dissolving, melting, granulating, dragee-making, tabletting, suspending, extruding, spray-drying, levigating, emulsifying, (nano/micro-) encapsulating, entrapping, or lyophilization processes. As noted above, the compositions of the present invention can include one or more physiologically acceptable inactive ingredients that facilitate processing of active molecules into preparations for pharmaceutical use.

Proper formulation is dependent upon the desired route of administration. For intravenous injection, for example, the composition may be formulated in aqueous solution, if necessary using physiologically compatible buffers, including, for example, phosphate, histidine, or citrate for adjustment of the formulation pH, and a tonicity agent, such as, for example, sodium chloride or dextrose. For transmucosal or nasal administration, semisolid, liquid formulations, or patches may be preferred, possibly containing penetration enhancers. Such penetrants are generally known in the art. For oral administration, the compounds can be formulated in liquid or solid dosage forms and as instant or controlled/sustained release formulations. Suitable dosage forms for oral ingestion by a subject include tablets, pills, dragees, hard and soft shell capsules, liquids, gels, syrups, slurries, suspensions, and emulsions. The compounds may also be formulated in rectal compositions, such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.

Solid oral dosage forms can be obtained using excipients, which may include, fillers, disintegrants, binders (dry and wet), dissolution retardants, lubricants, glidants, antiadherants, cationic exchange resins, wetting agents, antioxidants, preservatives, coloring, and flavoring agents. These excipients can be of synthetic or natural source. Examples of such excipients include cellulose derivatives, citric acid, dicalcium phosphate, gelatine, magnesium carbonate, magnesium/sodium lauryl sulfate, mannitol, polyethylene glycol, polyvinyl pyrrolidone, silicates, silicium dioxide, sodium benzoate, sorbitol, starches, stearic acid or a salt thereof, sugars (i.e. dextrose, sucrose, lactose, etc.), talc, tragacanth mucilage, vegetable oils (hydrogenated), and waxes. Ethanol and water may serve as granulation aides. In certain instances, coating of tablets with, for example, a taste-masking film, a stomach acid resistant film, or a release-retarding film is desirable. Natural and synthetic polymers, in combination with colorants, sugars, and organic solvents or water, are often used to coat tablets, resulting in dragees. When a capsule is preferred over a tablet, the drug powder, suspension, or solution thereof can be delivered in a compatible hard or soft shell capsule.

In one embodiment, the compounds of the present invention can be administered topically, such as through a skin patch, a semi-solid or a liquid formulation, for example a gel, a (micro)-emulsion, an ointment, a solution, a (nano/micro)-suspension, or a foam. The penetration of the drug into the skin and underlying tissues can be regulated, for example, using penetration enhancers; the appropriate choice and combination of lipophilic, hydrophilic, and amphiphilic excipients, including water, organic solvents, waxes, oils, synthetic and natural polymers, surfactants, emulsifiers; by pH adjustment; and use of complexing agents. Other techniques, such as iontophoresis, may be used to regulate skin penetration of a compound of the invention. Transdermal or topical administration would be preferred, for example, in situations in which local delivery with minimal systemic exposure is desired.

For administration by inhalation, or administration to the nose, the compounds for use according to the present invention are conveniently delivered in the form of a solution, suspension, emulsion, or semisolid aerosol from pressurized packs, or a nebuliser, usually with the use of a propellant, e.g., halogenated carbons derived from methane and ethane, carbon dioxide, or any other suitable gas. For topical aerosols, hydrocarbons like butane, isobutene, and pentane are useful. In the case of a pressurized aerosol, the appropriate dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of, for example, gelatin, for use in an inhaler or insufflator, may be formulated. These typically contain a powder mix of the compound and a suitable powder base such as lactose or starch.

Compositions formulated for parenteral administration by injection are usually sterile and, can be presented in unit dosage forms, e.g., in ampoules, syringes, injection pens, or in multi-dose containers, the latter usually containing a preservative. The compositions may take such forms as suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulatory agents, such as buffers, tonicity agents, viscosity enhancing agents, surfactants, suspending and dispersing agents, antioxidants, biocompatible polymers, chelating agents, and preservatives. Depending on the injection site, the vehicle may contain water, a synthetic or vegetable oil, and/or organic co-solvents. In certain instances, such as with a lyophilized product or a concentrate, the parenteral formulation would be reconstituted or diluted prior to administration. Depot formulations, providing controlled or sustained release of a compound of the invention, may include injectable suspensions of nano/micro particles or nano/micro or non- micronized crystals. Polymers such as poly(lactic acid), poly(glycolic acid), or copolymers thereof, can serve as controlled/sustained release matrices, in addition to others well known in the art. Other depot delivery systems may be presented in form of implants and pumps requiring incision.

Suitable carriers for intravenous injection for the molecules of the invention are well-known in the art and include water-based solutions containing a base, such as, for example, sodium hydroxide, to form an ionized compound, sucrose or sodium chloride as a tonicity agent, for example, the buffer contains phosphate or histidine. Co-solvents, such as, for example, polyethylene glycols, may be added. These water-based systems are effective at dissolving compounds of the invention and produce low toxicity upon systemic administration. The proportions of the components of a solution system may be varied considerably, without destroying solubility and toxicity characteristics. Furthermore, the identity of the components may be varied. For example, low-toxicity surfactants, such as polysorbates or poloxamers, may be used, as can polyethylene glycol or other co-solvents, biocompatible polymers such as polyvinyl pyrrolidone may be added, and other sugars and polyols may substitute for dextrose.

For composition useful for the present methods of treatment, a therapeutically effective dose can be estimated initially using a variety of techniques well-known in the art. Initial doses used in animal studies may be based on effective concentrations established in cell culture assays. Dosage ranges appropriate for human subjects can be determined, for example, using data obtained from animal studies and cell culture assays.

A therapeutically effective dose or amount of a compound, agent, or drug of the present invention refers to an amount or dose of the compound, agent, or drug that results in amelioration of symptoms or a prolongation of survival in a subject. Toxicity and therapeutic efficacy of such molecules can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., by determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dose ratio of toxic to therapeutic effects is the therapeutic index, which can be expressed as the ratio LD50/ ED50. Agents that exhibit high therapeutic indices are preferred.

The effective amount or therapeutically effective amount is the amount of the compound or pharmaceutical composition that will elicit the biological or medical response of a tissue, system, animal, or human that is being sought by the researcher, veterinarian, medical doctor, or other clinician, e.g., treatment of cancer, including induction of anti-tumor effects, etc. Dosages preferably fall within a range of circulating concentrations that includes the ED50 with little or no toxicity. Dosages may vary within this range depending upon the dosage form employed and/or the route of administration utilized. The exact formulation, route of administration, dosage, and dosage interval should be chosen according to methods known in the art, in view of the specifics of a subject's condition.

Dosage amount and interval may be adjusted individually to provide plasma levels of the active moiety that are sufficient to achieve the desired effects, i.e., minimal effective concentration (MEC). The MEC will vary for each compound but can be estimated from, for example, in vitro data and animal experiments. Dosages necessary to achieve the MEC will depend on individual characteristics and route of administration. In cases of local administration or selective uptake, the effective local concentration of the drug may not be related to plasma concentration.

In some embodiments of the present invention, effective doses for compounds of the invention include doses of 1 mg/kg, 2 mg/kg, 3 mg/kg, 4 mg/kg, 5 mg/kg, 6 mg/kg, 7 mg/kg, 8 mg/kg, 9 mg/kg, 10 mg/kg,

15 mg/kg, 20 mg/kg, 25 mg/kg, and 30 mg/kg, respectively.

In additional embodiments, effective treatment regimes for compounds of the invention include administration two or three times weekly.

The amount of agent or composition administered may be dependent on a variety of factors, including the sex, age, and weight of the subject being treated, the severity of the affliction, the manner of administration, and the judgment of the prescribing physician.

The present compositions may, if desired, be presented in a pack or dispenser device containing one or more unit dosage forms containing the active ingredient. Such a pack or device may, for example, comprise metal or plastic foil, such as a blister pack, or glass and rubber stoppers such as in vials. The pack or dispenser device may be accompanied by instructions for administration. Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of an indicated condition.

These and other embodiments of the present invention will readily occur to those of ordinary skill in the art in view of the disclosure herein. EXAMPLES

The invention is further understood by reference to the following examples, which are intended to be purely exemplary of the invention. The present invention is not limited in scope by the exemplified embodiments, which are intended as illustrations of single aspects of the invention only. Any methods that are functionally equivalent are within the scope of the invention. Various modifications of the invention in addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications fall within the scope of the appended claims.

Example 1: Increased Neural Progenitor CeU Proliferation and Viability

The effect of compounds and methods of the present invention on mesencephalic and cortical human neural progenitor cell (hNPC) proliferation and viability was examined using an MTT cell viability assay as follows. Human mesencephalic neural progenitor cell (hmNPC) and human cortical neural progenitor cell (frontal, hfNPCs) cultures were derived from tissue obtained from a 10-14-week-old human fetus. Two-midbrain-derived hNPC (hmNPC M 1 and hmNPC M2) and one frontal-derived hNPC (hfNPC F 1 ) were used in this MTT cell viability assay. The hNPCs were propagated in an undifferentiated state using methods previously described. (Milosevic et al. (2007) Stem cells Dev 16:625-635.) Briefly, expansion of hNPCs was carried out in a monolayer via plating the hNPCs onto polyornithine-fibronectin pre-coated culture dishes at a density of 30,000 cells / cm ² in reduced atmospheric oxygen (3 %). hNPCs were expanded in defined media (expansion media) (DMEM/F12) supplemented with epidermal growth factor

(EGF, 20 ng/ml), fibroblast growth factor-2 (FGF-2; 20 ng/ml, both from PromoCell, Heidelberg, Germany) and 2% B-27 (Invitrogen, Carlsbad, CA).

Neural progentitor cell proliferation and viability were measured using a 3-(4,5-dimethyl-2-thiazolyl)-2,5- diphenyl-2H-tetrazolium (MTT) assay essentially as described previously. (Mossman (1983) J Immunol

Methods 65:55-63; Sabolek et al. (2008) Neurotoxigology 29:714-721.) In this assay, hNPCs obtained and cultured as described above were seeded into 96-well culture plates at a density of 20,000 cells/well (in 100 μl medium), cultured for 24 hours in expansion media in 21% oxygen and then treated with various concentrations of Compound A (0, 5, 10, 20, and 30 μM, Figure IA), deferoxamine (DFO; 0, 10, 50, 100, and 200 μM, Figure IB), ciclopirox olamine (CPX; 0, 1, 5, 10, and 20 μM, Figure 1C), cobalt chloride (CoCl ₂ ; 0, 10, 25, 50, and 100 μM, Figure ID), or dimethyloxallyl glycine (DMOG; 0, 50, 100, 500, and 1000 μM, Figure IE) in expansion media for 72 hours.

After 72 hours, 10 μl of MTT reagent (5 mg/ml MTT) was added to each well and the cultures were incubated for an additional 3 hours. The resulting formazan dye was extracted with acid-isopropanol

(0.04 M HCl in absolute isopropanol) and the absorbence was measured spectrophotometrically with a computer-operated immuno reader (Tecan Deutschland GmbH, Crailsheim, Germany) at a wavelength of 570 nm with reference at 630 ran. The resulting values are expressed as percentage cell viability compared to that observed in untreated control cultures, and represent mean +/- SEM from 6 independent experiments.

As shown in Figure IA, Compound A significantly and dose-dependently improved hNPCs viability.

The effect of Compound A at increasing tiNPC viability was observed in each of the three hNPC cell preparations examined (hmNPC Ml, hmNPC M2, and hfNPC Fl). In particular, addition of Compound A at 5 μM and 10 μM increased the viability of hmNPC M2 by 35.8 ± 3.6% and 38.6 ± 1.7%, respectively. This result indicated that compounds and methods of the present invention are affective at increasing neural progenitor cell viability.

The effect of two different iron chelators (DFO and CPX, known to stabilize HIF- lα) on hNPC viability was also examined. Addition of DFO and CPX reduced neural progenitor cell numbers in all three hNPC preparations examined. In particular, addition of DFO decreased the number of viable hmNPC Ml by 22 ± 2% at 10 μM, by 36 ± 2% at 50 μM, by 60 ± 2% at 100 μM and by 65 ± 4% at 200 μM.

(See Figure IB.) CPX addition significantly reduced the number of hNPC in all three cell preparations examined (P < 0.001, Figure 1C) and, in particular, reduced cell viability in hmNPC Ml by 56.3 ± 1.7% at 1 μM. As shown in Figure ID, cobalt chloride did not significantly influence cell viability within the 72 hour incubation period. As shown in Figure IE, 0.5 mM DMOG reduced viability by 33 ± 2.6% in hmNPC M2 and by 54.2 + 1.5% in hmNPC M 1.

In a similar series of experiments, significantly more cells (higher numbers of living cells) were observed in hmNPC cultures grown for 1 week in 3% oxygen compared to that observed in hmNPC cultures grown for 1 week in 21% oxygen (P < 0.01, n = 6). (See Figure 2.) This result indicated that low oxygen conditions (e.g., hypoxia) increased hmNPC cell number and proliferation. Increased hmNPC cell numbers were observed in hmNPCs cultured in 21% oxygen (e.g., normoxia) in the presence of Compound A (5 μM or 10 μM) for 1 week, similar to that observed in hmNPC cultures grown in 3% oxygen (Figure 2). These results indicated that compounds and methods of the present invention increased neural cell proliferation and, in particular, increased neural progenitor cell proliferation.

Example 2: Regulation of Cell Cycle

The effect of compounds and methods of the present invention on neural cell cycle was examined as follows. hmNPCs were cultured and expanded in 21% oxygen as described above in Example 1, and cell cycle phase distribution of the cells was determined as follows. DNA content of the cells, as reflected by the fluorescence signal of propidium iodide, was measured using a flow cytometer (Becton Dickinson,

Heidelberg, Germany). Control and compound-treated hmNPCs samples were prepared for cell cycle analysis by lysing the cells in 300 μl of hypotonic fluorescence solution (HFS) as previously described. (See Milosevic et al., (2007) Stem Cells Dev 16:625-635 and Nicoletti et al. (1991) J Immunol Methods 139:271-279.) Histograms of DNA content were acquired using CellQuest software (Becton Dickinson). The number of nuclei present in each peak of the histogram: left to the Gl (sub-Gl), G1/G0, S, G2/M was analyzed by measuring the peak area using ModFit LT software (Verity, Turramurra, Australia).

One week incubation of expanding hmNPCs in the presence of various concentrations of Compound A (in the absence of B-27) increased cell cycle S-phase distribution. In particular, S-phase cell cycle distribution of hmNPCs increased from 8 + 0.1% in control cultures to 10 ± 0.6% in hmNPCs treated with 20 μM Compound A. (See Figure 3.) These results indicated that methods and compounds of the present invention are effective at modulating cell cycle in neural progenitor cells, and, in particular, increasing cell cycle S-phase distribution.

Example 3: Increased Proliferation and Neurons in hmNPCs

The effect of methods and compounds of the present invention on neural progenitor cell proliferation and neurogeneis was examined as follows. hmNPCs were allowed to differentiate in vitro for 2 weeks.

Differentiation of the cells was induced via replacement of expansion media by defined mitogen-free media supplemented with 2% B-27 and 5 μM forskoline (Sigma, St. Louis, MO).

Proliferating hNPCs were identified by double-immunostaining for TUJl and the proliferation marker Ki67 as follows. Control and compound-treated hmNPCs (from at least 2 different tissue preparations) were fixed in 4% paraformaldehyde in PBS for 15 minutes at room temperature, rinsed with PBS, counterstained with the DNA-binding dye 4'-6-Diamidino-2-phenylindole (DAPI, 2 μg/ml in PBS) for 15 minutes at room temperature, then twice rinsed in PBS followed by incubation in blocking buffer (10% FCS, 0,2% Triton-X 100 in PBS, pH 7.2) for 30 minutes at room temperature. After incubation with anti-β-iπ- tubulin primary antibody (TUJl ; Covance, Berkeley,CA, USA), anti-Ki67 antigen antibody (Novocastra Laboratories Ltd, Newcastle upon Tyne, UK), or anti-doublecortin (DCX) primary antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA) for 1 hour at room temperature in blocking buffer, the cells were incubated with Alexa Fluor® 488 conjugated or Alexa Fluor® 594 conjugated secondary antibodies (Molecular Probes, Eugene, USA). Coverslips were mounted onto the glass slides and staining examined under a fluorescence microscope (Zeiss Axiovert 200). Acquisition of the immunostained cells was performed using the Image-analysis software Axio Vision 4 (Carl Zeiss AG, Jena, Germany).

Neurogenesis (e.g., increase in neural cell proliferation, increase in newly generated neurons, and increase in immature neurons) was identified by the presence of two characteristic and identifying neural cell markers: neuron-specific class UJ β-tubulin (TUJl, immunostained using anti-β-III- tubulin primary antibody) and doublecortin (DCX, immunostained using anti-doublecortin (DCX) primary antibody, Santa Cruz Biotechnology, Inc., Santa Cruz, CA). TUJl is expressed in neurons of the peripheral and central nervous systems and contributes to microtubule stability in neuronal cell bodies and axons. DCX is a microtubule-associated protein expressed almost exclusively in immature (e.g., developing) neurons, and thus is a marker for neurogenesis. Neural precursors begin to express DCX shortly after exiting the cell cycle, and continue to express DCX as the cells mature into neurons. (Couillard-Despres et al. (2005)

Eur J Neurosci 21:1-14.)

Proliferating neural cells were identified by double immunostaining for TUJl (green in Figures 4A, 4B, and 4C) and Ki67 (red in Figures 4 A, 4B, and 4C). As shown in Figures 4A, 4B, and 4C, arrows indicate proliferating neuroblasts (TUJl ⁺ /Ki67 ⁺ ) in one representative hmNPC preparation. Virtually all TUJl ⁺ cells were also positive for DCX (Figure 4A, insert). Neural cell nuclei are shown in blue (DAPI stained). Scale bar = 50 μm.

The number of hNPCs positive for TUJl ⁺ , Ki67 ⁺ , or both markers (TUJl ⁺ /Ki67 ⁺ ) were determined in control and Compound A-treated cultures and normalized to the total number of DAPI-stained cells. The percentage of immature neurons increased from 5.8 ± 0.9% in non-treated control NPC cultures to 11.1 ± 0.7% obtained in NPC cultures treated with 10 μM Compound A (one-way ANOVA, P < 0.001, n = 6). (See Figure 5.)

The number of expanding neural cells significantly increased following addition of Compound A, as revealed by Ki67 staining (P = 0.016), with the highest expression of Ki67 in 10 μM Compound A vs. control (23.6 ± 2.9% vs. 12.0 ± 0.9%, respectively). TUJl ⁺ /Ki67 ⁺ cells were rare to find in untreated cultures, but in cultures treated with either 5 μM and 10 μM Compound A, double-positive cells were quantified as 1.0 ± 0.1% and 1.2 ± 0.1%, respectively (P < 0.01; Figure 5). In Figure 5, the numbers of immunopositive cells (Ki67 ⁺ only, TUJl ⁺ only, or Ki67 ⁺ /TUJl ⁺ ) expressed as the percentages of DAPI stained nuclei are shown as mean ± SEM (D). *P < 0.05 when compared to untreated control (n = 6). These results indicated that compounds and methods of the present invention are effective at increasing neural cell proliferation and neurogenesis.

Example 4: Increased Neural and Dopaminergic Differentiation

The effect of methods and compounds of the present invention on differentiation of neural cells was examined as follows. Mesencephalic NPCs were differentiated for one week in 21% oxygen with simultaneous exposure to Compound A (5 μM and 10 μM). Following this treatment, the cells were fixed and immunostained for the mature neural cell marker microtubule-associated protein-2 (MAP2) using anti-microtubule associated protein-2 (MAP2ab; BD Pharmingen, San Diego, CA, USA) to identify mature neural cells, and immunostained for the presence of tyrosine hydroxylase (TH) using rabbit polyclonal anti-tyrosine hydroxylase (anti-TH) antibody (Santa Cruz) to identify the presence of dopaminergic neurons (Figures 6 A and 6B).

A representative human midbrain-derived NPC was immunostained for the neuronal marker microtubule- associated protein 2 (MAP2) and the dopaminergic marker tyrosine hydroxylase (TH) upon 1 week of differentiation in 21% oxygen in the presence or absence of 10 μM Compound A. As shown in Figures 6A and 6B, administration of Compound A increased TH and MAP2 expression in neural cells. In Figures 6A and 6B, TH is red, MAP2 is green, and DAPI is blue.

Immunoblotting was performed as follows. Combined cytoplasmic and nuclear extracts of cultured hmNPCS were prepared in extraction buffer as described previously (Milosevic et al., 2007a). Protein concentrations were determined by the Bradford method, using bovine serum albumin as a standard. Denaturated proteins were resolved on a sodium dodecyl sulfate (SDS)-polyacrylamide gels and transferred to a Hybond-ECL nitrocellulose membrane (GE Healthcare, Freiburg, Germany) by semidry blotting. Membranes were stained with Ponceau S (Sigma) to confirm equal protein loading and transfer followed by blocking with 5% (wt/vol) nonfat dry milk in PBS-T (PBS, 0.1% (vol/vol) Tween 20) for ^• 2 hours at room temperature and subsequent incubation with desired primary antibody (diluted in PBS containing 5% non-fat milk and 0.1% Tween 20) overnight at 4 ⁰ C with gentle agitation. The antibodies used were as follows: mouse monoclonal anti-actin (C4, ICN Biomedicals), rabbit polyclonal anti- tyrosine hydroxylase (anti-TH) antibody (Santa Cruz), mouse monoclonal anti-TUJl (Covance) mouse monoclonal anti-neuron specific enolase (NSE, Chemicon International, Hampshire, UK), horseradish peroxidase-coupled secondary antibodies (Pierce, Rockford, IL, USA). Chemiluminescence detection was performed by incubating the membranes with SuperSignal-Dura substrate (Pierce) followed by analyzing on a CCD cooling camera (Fuji LAS-1000plus). The chemiluminescence was quantified using AIDA, two-dimensional densitometry software (Raytest Isotopenmeb, Straubenhardt, Germany).

The numbers of immunopositive cells (TH ⁺ cells) expressed as the percentages of DAPI stained nuclei are counted and presented as mean ± SEM.

For each sample, untreated and treated, 10 randomly selected fields with approximately 300-500 DAPI ⁺ nuclei were chosen and TH ⁺ neurons were counted. Dopaminergic differentiation was then determined and is expressed as a percentage OfTH ⁺ ZDAPI ⁺ cells. This ratio was 1.2 ± 0.1% in untreated samples and increased to 2.5 ± 0.3% and 3.0 ± 3% in cells treated with 5 μM and 10 μM Compound A, respectively. (Figure 7). Treatment of hmNPCs with Compound A during differentiation significantly increased dopaminergic differentiation of the cells, as measured by increased TH+ expression (one-way ANOVA; P

< 0.001; n = 10). Multiple comparisons versus the untreated control cells by Tukey test revealed a significant effect of 10 μM Compound A on dopaminergic differentiation of these cells. Figure 8 shows mmunoblots representing alterations in the expression of neuronal (NSE) and dopaminergic (TH) markers obtained in hmNPCs following differentiation of NPC cultures in the presence of Compound A. As shown in Figure 8, immunoblots of both mature (as determined by neuron- specific enolase (NSE) reactivity) and dopaminergic neuron (TH) markers further confirmed their increased expression following treatment of NPC cultures with Compound A (Figure 8).

Taken together, these results indicated that compounds and methods of the present invention are effective at increasing neural and dopaminergic differentiation.

Example 5: Increased Neurite Outgrowth

The effect of compounds and methods of the present invention on neurite outgrowth was examined as follows, using methods similar to those previously described. (See Shingo et al. (2001) J Neruosci

21:97339743 and Caldwell et al. (2001) Nat Biotechnol 19:475-479.) Neural progenitor cells were obtained from Lonza (Poietic NHNP Cells PT-2599 NHNP Neural Progenitor cells). The cells were cultured in media (Maintenance Medium NPMM, Lonza) containing fibroblast growth factor (FGF) and epidermal growth factor (EGF).

A series of experiments were performed to examine the effects of methods and compounds of the present invention on neural progenitor cell survival, expansion, and differentiation in both an expansion phase and a differentiation phase. In the expansion phase experiments, neural progenitor cells were cultured under growth conditions that support self-renewal and proliferation of non-differentiated cells, but which do not support spontaneous differentiation of the neural progenitor cells. In the differentiation phase experiments, neural progenitor cells were cultured under growth conditions that support differentiation of neural progenitor cells into mature neural cells.

Cells were cultured in the Maintenance Medium NPMM, (CC-3209 Lonza) containing FGF, EGF, and various concentrations of Compound B for three days, during which time neurospheres formed. The cells were then re-suspended, counted, and equivalent numbers of cells were plated onto laminin-coated wells (R&D Systems) in the absence of Compound B, EGF, and FGF (differentiation phase). The cells were then plated onto laminin coated slide chambers (60,000 cells per /well), and cultured in NPDM™ - Neural Progenitor Differentiation media (CC-3229, Lonza).

As shown in Figures 9A and 9B (showing results of two separate experiments, respectively), addition of Compound B (3 μM or 6 μM) to neural progenitor cell cultures increased the percent of neurospheres containing neurites compared to non-treated control cultures. (Data in Figures 9A and 9B represent averages and standard deviations.) As shown in Figures 1OA and 1OB (showing results of two separate experiments, respectively), addition of Compound A (3 μM or 6 μM) to neural progenitor cell cultures increased the number of neurites per cell well compared to non-treated control culture well. (Data in Figures 1OA and 1OB represent averages and standard deviations.) The majority of cells containing neurites were positive for microtubule associated protein-2 (MAP2, by immunostaining, data not shown), indicating differentiation of the neural progenitor cells to a neural cell fate. Taken together, these results indicated that compounds and methods of the present invention are effective at increasing neurites in neural progenitor cells.

Example 6: Increased VEGF Gene Expression in Neural Progenitor Cells

The effect of compounds and methods of the present invention on expression of vascular endothelial cell growth factor (VEGF) in neural progenitor cells was examined as follows. Neural progenitor cells obtained from Lonza (see Example 5 above) were cultured in mainteinance media for 48 hours in 96-well culture plates in the presence or absence of Compound B (3 μM or 10 μM). Messenger RNA from the cells was isolated using an mRNA purification kit (RNeasy 96 Kit, QIAGEN #74182) and analyzed for

VEGF expression (Gene Expression Assay, 00173626 ml, Applied Biosystems).

As shown in Figure 11 , Compound B increased VEGF expression in the neural progenitor cells compared to that observed in non-treated control cultures. These results indicated that methods and compounds of the present invention are effective at increasing VEGF expression in neural progenitor cells.

Example 7: Increased Neurogenesis in vivo

The effect of compounds and methods of the present invention on neurogenesis in vivo was examined as follows. Male C57B1/6 mice (8-10 weeks old; Charles River Laboratories) were singly housed under standard conditions and provided with food and water ad libitum. Animals were randomly divided into 4 groups of 8 animals each, and groups were treated 3 times per week for 3 weeks (10 treatments) with either vehicle control, Compound A (60 mg/kg), Compound C (60 mg/kg), or Compound D (60 mg/kg) by oral delivery. On the day after the final dose, all groups received two intraperitoneal injections with 5-bromo-2-deoxyuridine (BrdU; 100 mg/kg for each injection) with a two hour interval to label actively dividing cells. Three weeks after the BrdU treatment, animals were deeply anaesthetized using isoflurane and tissues were perfused with saline followed by 4% cold paraformaldehyde by transcardiac perfusion. Brains were removed and post-fixed overnight in 4% paraformaldehyde at 4 ⁰ C, trimmed to remove excess rostral and caudal brain tissue, and then cryoprotected in 30% sucrose. Brains were coronally embedded in TISSUE-TEK OCT (Optimal Cutting Temperature) compound (Sakura) and stored at -80 ⁰ C. Brains from 4 animals per treatment group were serially sectioned into 35 um sections on a cryostat and stored in

PBS with 0.1% sodium azide. Every 4th section was subjected to immunohistochemistry in parallel for detection of BrdU and NeuN (Neuronal Nuclei). Briefly, sections were rinsed in PBS without azide before proceeding with the following steps: 1 hr incubation in 50% formamide/2X SSC solution at 65 ⁰ C with gentle agitation; 5 min rinse in 2X SSC at room temperature; 30 min incubation in 2N HCl at 37 ⁰ C; 10 min rinse in 0.1M boric acid. Sections were blocked for 30 min in Tris Buffered Saline (TBS) supplemented with 0.1% TritonX-100 and 5% normal donkey serum; sections were incubated overnight with primary antibodies (BrdU 1 : 100, Serotec; NeuN 1 :500, Chemicon) diluted in blocking buffer at 4°C.

The next day, sections were rinsed 3 times in TBS before application of secondary antibodies: donkey anti-mouse Cy3 (1:1000) and biotin-conjugated donkey anti-rat (1: 100) followed by streptavidin-Cy2 (1:200) all from Jackson ImmunoResearch. Sections were mounted on glass slides and coverslipped prior to examination. Sections were first analyzed for nuclear BrdU staining pattern at 2Ox magnification, and then uniform and punctate BrdU-positive cells in or immediately adjacent to the subgranular zone were counted using 6Ox magnification.

The percentage of cells exhibiting BrdU in a punctate pattern, which indicates clumping of DNA into heterochromatin (see, e.g., Cameron and McKay (2001) J Comp Neurol 435:406-417), was also calculated to determine the degree to which neurogenesis resulted in mature neurons. As shown in Figure

12A, mice treated with compounds and methods of the present invention had an increased percentage of new mature neurons relative to vehicle-treated controls. These results indicated that methods and compounds of the present invention are effective at increasing neurogenesis in vivo.

In order to assess the functional significance of increased hippocampal neurogenesis, male C57BL/6NCrl mice (28 day-old; Charles River Laboratories) were treated every other day for 3 weeks (11 treatments) with either vehicle or Compound A (60mg/kg) by intraperitoneal injection. Groups of animals were examined at either 1, 3 or 4 weeks after the final treatment to test for hippocampal-dependent contextual fear conditioning as described by Adamcio et al. (2008) BMC Biol 6:37. In brief, after a 120s period in which baseline freezing was assessed, mice received a 2s, 0.4mA foot shock. Contextual memory was assessed 72 h after the training by monitoring mice over a 2 min period for freezing in the same conditioning chamber. Statistical significance was evaluated using 2-tailed unpaired t-test and two-way ANOVA. Significance level was set to p<0.05. Numerical values are presented as mean±S.E.M. in Figures and text. Plotting of the data and statistical analyses was done in Prism 4 (GraphPad Software, San Diego, CA, USA).

As shown in Figure 12B, mice treated with compound A showed a clear improvement in contextual memory in fear conditioning at both 3 and 4 weeks after treatment cessation. Thus, the compounds and methods of the invention increased neurogenesis in the hippocampus and improved functional hippocampal-dependent contextual memory. Various modifications of the invention, in addition to those shown and described herein, will become apparent to those skilled in the art from the foregoing description. Such modifications are intended to fall within the scope of the appended claims.

All references cited herein are hereby incorporated by reference herein in their entirety.