Method for treating diabetic neuropathy with NGF

Drug – bio-affecting and body treating compositions – Designated organic active ingredient containing – Peptide containing doai

Reexamination Certificate

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C514S003100

Reexamination Certificate

active

06440928

ABSTRACT:

FIELD OF THE INVENTION
This invention generally relates to therapeutic processes and compositions for ameliorating damage to mammalian nerve cells, as well as for healing and/or repairing said nerve cells, especially in the case of neuropathy. In this context, the term “nerve cells” should be taken to include neurons and neuroglia, whether in the central or peripheral nervous system, including the pre- and post- synapaptic elements of synapses and neuromuscular junctions. Similarly, the term “repair” should be taken to include recruitment of new nerve cells as well as restoration of function of existing nerve cells. Similarly, the expression “functionally repairing” nerve cells should include the concepts of recruitment of new nerve cells as well as the restoration function of existing nerve cells. This invention is particularly concerned with processes and compositions wherein insulin and insulin-like growth factors play a significant therapeutic role in neuropathy, especially diabetic neuropathy.
BACKGROUND OF THE INVENTION
The biochemistry and physiology associated with amelioration of damage to nerve cells, and induced reparations to nerve cells (including nerve cell elements such as neurites, synapses, epineurium, and endoneurium) have been the subject of many years of investigation. The field is complex and is often characterized by the presence of many subtle—yet extremely significant—distinctions. Some of these distinctions are, in turn, very much influenced by the underlying theories and/or assumptions employed by any given worker in this field. For example, much of the reported work has been done in conjunction with studies of particular kinds of damage to nerve cells through physical trauma or pathological disorders such as diabetic neuropathy. Those skilled in this art will appreciate that the processes of repair are not necessarily the same in the case of trauma as in pathological disorders. A great deal has been learned and some useful therapies have been implemented. A review of some of this past work will be helpful in establishing the scope of the herein described processes for healing nerve cells.
To these ends, it first should be noted that it has long been established that diabetic autonomic neuropathy includes abnormal regulation of blood pressure, bowel function, sweating, and skin temperature. Sensory neuropathy is known to include the absence of deep reflexes and loss of ability to perceive stimuli such as pain and temperature. Moreover, various stages of degeneration in sympathetic ganglion cells have been observed in the presence of this disease state. It is also known that conduction velocity is often reduced in sensory, motor and probably sympathetic nerves. Moreover, the prior art has, to some degree, appreciated that such reduction is partially preventable by insulin treatment.
It also has long been established that axon loss and segmented demyelination may be present in peripheral and sympathetic nerves. While demyelination suggests an indirect effect due to alteration in Schwann cell activity, the decrease in conduction velocity in juvenile onset and experimental diabetes is, however, not necessarily accompanied by morphologic alterations of the myelin sheath. Indeed, axonal disease is also known to be present in unmyelinated fibers. Moreover, while distal symmetrical polyneuropathy is more commonly encountered than symmetrical proximal motor neuropathy, or focal and multifocal neuropathies, the various forms of neuropathy can be present together in the same patient. Therefore, classifications into these subtypes is clinically useful, but may not truly reflect distinct categories of neuropathies.
It should also be noted that, when viewed in total, the prior art has shown that insulin and their receptors are present in the central nervous system, but the prior art has not shown whether or not the receptors can actually regulate neurite formation in vivo. For example, applicant's hereinafter cited past studies with respect to the effects of insulin and IGFs were conducted in vitro. Indeed, many technical considerations and references have indicated that in vitro studies do not reveal whether insulin, IGF-I, or IGF-II are active on the nervous system in vivo. For example, cells placed in culture are in fact removed from their normal in vivo environment. In such cases, they are removed from a host of supportive factors. Moreover, the capacity of cells to respond to an exogenous factor in vitro does not necessarily imply that that particular factor would have significant activity in vivo, or that a similar physiological role is played in vivo. For example, with respect to the latter, it is known that neurite outgrowth can be induced in vivo by actinomycin D or bromodeoxyuridine. One may not, however, infer that these compounds are physiological regulators of neurite outgrowth. Cultured cells are also removed from in vivo inhibitory substances. Such factors have been found to be present on CNS oligodendrocytes, which might help explain why regeneration is so poor in the CNS (Schwab and Caroni, 1988, J. Neurosci. 8:2381-2393). Hence, it is important to understand the teachings of this patent disclosure to appreciate and demonstrate the effects of insulin and IGFs on nerve cells in vivo. This is important because such a showing makes a distinction between the teachings of this patent disclosure and those made in various prior art references such as the Hansson et al, Acta Physiol Scand. 126 (609-614) (1986) reference. More will be said about the teachings of the patent disclosure vis-a-vis the scope of the teachings of this 1986 Hansson reference, especially in view of a subsequent Hansson paper published in 1987.
Those skilled in this art also will appreciate that previous theories regarding neuropathy include the involvement of small vessel angiopathy, secondary response to disturbances in Schwann call function, excess production and accumulation of polyols through the sorbitol pathway, alteration in lipid metabolism, decreased myoinositol production, and abnormal glycosylation of proteins. Detailed discussions of these theories are available throughout the literature (see for example (1) Thomas and Eliasson, In “Peripheral Neuropathy”, Dyck et al., eds. WB Saunders Co., Philadelphia, 1984 pp 1773-1810 and (2) Brown and Greene, In “Peripheral nerve disorders, a practical approach” Asbury and Gilliatt, eds., Buttersworth, London, 1984 pp 126-53); but there is by no means any consensus as to the pathogenesis or extent of pathology (see Powell, Lab Invest 49:515, 1983).
In any event, none of the various prior art theories have thus far fully considered that insulin may act directly on neurons, or that IGFs, protein kinase C, and various neurotrophic factors such as nerve growth factor (NGF) may also play roles in the pathogenesis of such disorders. Moreover, in those instances where insulin was considered, no clear conclusions could be drawn. For example, Applicant and his coworkers previously have shown that physiological concentrations of insulin can reversibly increase the proportion of cells with neurites, as well as average neurite length, in cloned human neuroblastoma SH-SY5Y cells in culture (see, Recio-Pinto and Ishii, Brain Res 302:323-334, 1984). Other workers have established that this response can be inhibited by anti-insulin antiserum, and that it is correlated with occupancy of high affinity insulin receptors. It also has been established that insulin can directly enhance neurite formation and support neuron survival in primary cultures of sensory and sympathetic cells (see, Recio-Pinto et al., J Neurosci 6:1211-1219, 1986). The presence of other serum factors, or non-neuronal cells, did not appear to be required for the response.
These past speculations, based on in vitro studies, that insulin plays a role in the development and maintenance of the peripheral and central nervous system also found some support in past observations showing that insulin induces the precocious maturation of evocable synaptic transmission in culture. It wa

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