CN102355902A - Treatment of neurotrophic factor mediated disorders - Google Patents

Abstract

An agent selected from A/B-cis furostane, furostene, spirostane and spirostene steroidal sapogenins and ester, ether, ketone and glycosylated forms thereof is used to induce self- regulated homeostasis of neurotrophic factors(NFs), for example BDNF and/or GDNF, NFs with limited and manageable side effects in a subject, by modulating NFs in a non- toxic manner under homeostatic control. An effective amount of at least one such agent is administered to the subject, particularly in the treatment or prevention of a range of NF- mediated disorders, particularly neurological, psychiatric, inflammatory,allergic,immune and neoplastic disorders, and in the restoration or normalisation of neuronal and other function in or in relation to any damaged or abnormal tissue, including when assisting tissue (for example, skin, bone, eye and muscle) healing and general skin, bone, eye and muscle health.

Description

The processing of the disease of neurotrophic factor mediation

Invention field

The present invention relates to the disease of neurotrophic factor mediation; Especially the processing and the prevention of neurological, psychiatry, inflammatory, allergia, immunity and tumprigenicity disease; With the neuron in any infringement or unusual tissue or relevant with any infringement or unusual tissue and the recovery or the normalization of other functions; Comprise when aid in tissue (for example skin, bone, eye and muscle) healing and general skin, bone, eye and muscle health, relate to relevant non-Therapeutic Method and relate to wherein used chemical compound and compositions.

Background of invention

Neurotrophic factor (NF) comprises neurotrophin, TGF-β-superfamily, NF and neurokine, and for example nerve growth factor (NGF), brain come the neurotrophic factor (GDNF) in derived neurotrophic factor (BDNF), ciliary ganglion neurotrophic factor (CNTF), neurotrophin 3 (NT-3), neurotrophin 4 (NT-4) and neuroglia source.Neurotrophic factor combines with the cell receptor that is known as neurotrophic factor acceptor (NFr).The effect of NFr TrkA mediation NGF.NFr TrkB is activated by BDNF, NT-3 and NT-4.NFr TrkC is activated by NT-3 only.The low affinity NGF receptor (LNGFR or p57) of NFr combines all members of neurotrophin family.The NFr of GDNF is grouped into by two kinds of one-tenth, GDNF binding structural domain (GDNF receptor alpha 1 (GFR α 1) and receptor tyrosine composition Ret.GDNF and GFR α 1 combine activation Ret.

The unusual expression of natural NF relates to many diseases, and on the basis of NF simulation that the non-peptide therapeutics of micromolecule generally acknowledged inferred or activating activities, has designed therapy.In principle; The non-peptide of micromolecule (comprising non-polypeptide and non-albumen) therapeutic agent has the many advantages above the peptide agent usually, comprise the low cost compared with peptide and relative simply preparation, simpler handle and storage, the intrinsic toxicity that reduces, relatively simply to patient especially sending and relatively simply optimize to brain in the research and development stage.Although peptide NF, NF-plan thing and NF reinforcing agent to as potential drug are very interested, have been found that their inherent exploitation difficulties, potential toxicity and other problem seriously limit their potentiality.

A large amount of non-peptides of micromolecule have been proposed to be used in handles some neurological and psychiatric disease.The following passage has been stressed some in the publication.Yet; Suggestion before these all has the characteristic that there is remarkable adverse side effect in agent; This hinders using of effective dose, makes that chemical compound can not be by exploitation to provide the medicine of introducing to the market of handling or preventing neurological and psychiatric disorders in all examples.

For example, and xaliproden (Sanofi-Aventis) (1-(2-naphthalene-2-base ethyl)-4-[3-(trifluoromethyl) phenyl]-3,6-dihydro-2H-pyridine hydrochloride; The MW:417.5 of salt; The MW:381 of free alkali), a kind of serotonin 5-HT _1AReceptor stimulating agent is found recently and also activates NGF to a certain extent.Xaliproden be in the news as the potential therapy of amyotrophic lateral sclerosis (ALS) has been accomplished III clinical trial phase (Drugs R D.2003,4 (6), 386-388 page or leaf) and test in the III phase as the potential therapy of Alzheimer recently in assess.Yet, 5-HT _1AAgonist activity produces the detrimental effect of dose dependent, and it has limited the use of xaliproden as medicine.

4-methyl catechol (MW 124) has been in the news and has excited nerve the synthetic of nutrient protein and a kind of method of handling neurodegeneration (people such as Furukawa is provided thus in theory; Advances in Behavioral Biology; 2002,53, the 233-236 page or leaf).Yet this a kind of dose has been found the toxigenicity side effect, maybe be owing to the excessive activation of nerve growth factor (NGF) expression.

Tretinoin has been in the news to be increased the serum of NGF and neural level and in diabetic mice, prevents neuropathy (people such as Arrieta; European Journal of Clinical Investigation; 2005; 35; The 201-207 page or leaf) and be indicated in the therapeutical effect (Mey and McCaffery, The Neuroscientist, 2004 that have possibility in the neurodegeneration disease; 10, the 409-420 pages or leaves).Yet this a kind of dose known has serious dose-limiting toxicity side effect.

Ampa receptor synergist (ampakine) is that glutamate receptor regulator and some have demonstrated and strengthen BDNF in vivo and express people such as (, Neuropharmacology, 2002,43, the 1-10 page or leaf) Mackowiak.And; Two kinds of ampakines (CX614 and CX546) have been presented to be used the back and increased BDNF mRNA level in 6-12 hour the biglyyest and be reduced near control level in back 48 hours using afterwards; Even successive ampakine contact (people such as Lauterborn; Journal of Pharmacology and Experimental Therapeutics; 2003; 307, the 297-305 pages or leaves).Several ampakines or are used for neurological's disease (people such as Price, Pharmacology and Therapeutics, 2007,115, the 292-306 page or leaf) by exploitation.Yet at least some in these agent have toxic side effects.

Some antidepressants; Comprise and having as those of the primary effect of serotonin selectivity reuptake inhibitor (SSRI) and oxidase inhibitor (MAOI); Shown and increased BDNFmRNA level (Malberg and Blendy in vivo; Trends in Pharmacological Sciences; 2005; 26, the 631-638 pages or leaves; People such as Martinez-Turrillas, Neuropharmacology, 2005,49,1178-1188 page or leaf).Also referring to Castren, Current Opinion in Pharmacology, 2004,4, the title of 58-64 page or leaf is the summary of " Neurotrophic effects of antidepressant drugs (neurotrophic effect of antidepressant drug) ".Yet all these agent all are known as has many side effect of not expecting.

Immunophilin (immunophillin) is to have shown to have to strengthen the active para-immunity inhibitor of neurotrophin people such as (, Pharmacology and Therapeutics, 2007,115, the 292-306 page or leaf) Price.FK506 (Tacrolimus) has shown increases BDNF mRNA level (Zawadzka and Kaminska in vivo; Molecular and Cellular Neuroscience; 2003; 22; The 202-209 page or leaf) and BDNF and gdnf protein level (people such as Tanaka, Brain Research, 2003; 970, the 250-253 pages or leaves).Yet whole class has serious dose-limiting toxicity side effect.

N4-(7-chloro-2-[(E)-2-(2-chloro-phenyl-vinyl)]-quinolyl-4)-N, N '-diethyl-pentane-1,4-diketone (XIB4035), a kind of GFR _*-1 receptor stimulating agent has been reported as with the concentration dependent mode and has promoted neurite outgrowth (people such as Tokugawa, Neurochemistry International, in January, 42,1,2003,81-86 page or leaf).Yet this a part also has the side effect of dose limitation property.

These known micromolecule agent have NF-simulation or activation to a certain degree thus, but preclinical models or clinical in have the adverse side effect of dose dependent.Side effect can show they self tangible toxicity usually.This has seriously limited the potential utility of agent described in the therapy.

Have and split hair improved in the therapy of neurological and psychiatric disorders, and especially avirulent, generally the needing of the non-peptide biological activity agent of micromolecule.

WO-A-99/16786, WO-A-99/48482, WO-A-99/48507, WO-A-01/23407, WO-A-01/23408, WO-A-02/079221, WO-A-03/082893, WO-A-2005/105108, WO-A-2005/105825 and WO-A-2006/048665; Its disclosure is incorporated this paper by reference into, relates to the purposes of some micromolecule steroid in the processing of cognitive dysfunction and some other neurological and psychiatric disorders.Generally speaking; These activating agents are A/B-cis furostan, furan steroid alkene (furostene), spirostane or spirostene (spirostene) steroid sapogenines and its ester, ether, ketone and glycosylated form; Statement " ruscogenin " is understood to include the derivant of all E and/or F ring opening, for example the pseudo-ruscogenin form of the pseudo-ruscogenin of said ruscogenin and dihydro.In unsaturated (alkene) form of chemical compound, one or more pairs of keys are present in the position that does not influence A/B cis primitive (motif).

WO-A-03/082893; The 25th page; In the said chemical compound of the 5th to 18 row report at least some have been found slows down or some aspect of reversing neuronal degeneration; Comprise and reverse disadvantageous cyton variation and neurite atrophy, reduce NF for example release and the minimizing neurotoxicity and the apoptosis of neurotrophin, TGF-β-superfamily NF and neurokine.This paragraph reports that also the neuroprotective of receptor forfeiture effect and counter-rotating are the effects of actively regulating, and wherein along with the prevention of continuous decline, the decline in past is reversed the state towards normal or initial stage.

Identical file, the 26th page, the 8th to 15 row also report it is believed that a kind of physiological effect of activating agent is the ability of the speed of the synthetic or release of increase NF or their receptor or the degraded that reduces NF or their receptor.In theory to these effects of somatomedin " maybe be owing to the effect of chemical compound to kytoplasm or nuclear receptor; perhaps chemical compound and promoter region combine and thereupon to the direct influence of the throughput rate of the mRNA of somatomedin, or increase the result of the production of the another kind of material factor ".

Identical file, the 20th page, the 4th has gone and has described the purposes that activating agent is handled lonely syndrome, depression and schizoid psychiatric disorders.

Zhang Y waits the people, FEBS Letters, and on March 19th, 2008,582, the 6 phases, the 956-960 page or leaf, its disclosure is incorporated this paper by reference into, and the report smilagenin shows to be increased with 1-methyl-4-phenylpyridinium (MPP ⁺) in the rat midbrain dopaminergic neuron of infringement GDNFmRNA express and culture medium in GDNF content, and smilagenin is presented in these neurons and suppresses MPP ⁺Inductive neuron infringement and atrophy.This publication is not a prior art from the inventor and at all designated states.

The effect of NF has been the theme (referring to for example Vega, people such as J A, J.Anat.2003,203, the 1-19 pages or leaves and the list of references of wherein being quoted, its all disclosure is incorporated this paper by reference into) of many researchs in the immune system stable state in recent years.As explain more in detail in people's such as Vega the publication with the 8th page table 2 in generalized, NF shown have with immune system in a large amount of cells of relating to especially bone-marrow-derived lymphocyte, T lymphocyte, monocyte/macrophage, neutrophil cell, oxyphil cell, basophilic granulocyte, mastocyte and hematopoietic cell and the platelet a large amount of activity relevant with vascular tissue.The steady-state adjustment of NF provides and can be used for handling or the of great value technology of epidemic prevention system disease.

The effect of NF in inflammation and inflammatory disease and the allergy also receives many concerns.Known inflammation and abnormal response duration and in immune disease the NGF level increase (referring to Stanisz, AM & Stanisz, JA, Ann.N Y Acad.Sci., 2000,917, the 268-272 page or leaf; Otten, people such as U, Ann.N.Y Acad.Sci, 2000,917,322-330 page or leaf; Also have people's such as Vega the 10th page of the 2nd row to be listed as the list of references of being quoted) with the 11st page the 1st and 2.The steady-state adjustment of NF provides and can be used for handling or prevention of inflammation and inflammatory disease and the abnormal of great value technology that responds.

Like called optical imaging, inflammatory, abnormality and immune response can take place simultaneously and with the mode of being mutually related, for example in the response that excites of autoimmune disease and contratoxin, parasite and other infectious agent.The steady-state adjustment of NF provides the of great value technology that can be used for handling or preventing these patient's condition.

NGF be presented at and had useful effect in the inductive rheumatoid arthritis of vasculitis (Tuveri, people such as M., Lancet, 2000Nov 18,356, the 1739-1740 page or leaf; Aloe, L., Arch.Physiol.Biochem., 2001,109, the 354-356 page or leaf) and report be considered to be in and stop during metamorphosis or the inflammatory process NF to cross new therapy strategy people such as (, the publication of above quoting, the 12nd page, the 1st row) Vega of expression.Because the reason of above being explained relevant with neurological's disease, the proteic use of NGF are not so good as more desirable of micromolecular use.Be used to regulate micromolecule agent that NF crosses expression and will be very desirable.

WO-A-01/64247, its disclosure is incorporated this paper by reference into, described handle or prevention by the NF receptor on cancerous cell surface especially trk ⁺The method of the tumprigenicity disease (cancer) that expression characterized on the cancerous cell.Method comprises the anti-NF agent (in list of references, being called as anti-neurotrophin or anti-NT agent) of using effective dose, for example anti-NF antibody, anti-NF antisense polynucleotides or anti-NF trk mutant.It is said a large amount of cancers; The cancer that comprises cancer, the brain cancer and the peripheral nervous system tissue of breast carcinoma, thyroid carcinoma, colon cancer, pulmonary carcinoma, ovarian cancer, skin carcinoma, muscle cancer, cancer of pancreas, carcinoma of prostate, renal carcinoma, anogenital cancer, leukemia, immune system tissue (for example spleen, thymus and bone marrow) can this method be handled or prevention.The mode of effect it is said it is to combine with the high specific of NF via activating agent, causes suppressing trk receptor (page 5, the 8th to 10 row) via the neutralization of activatory NF part.

Innominato, people such as P F, J.Pathol., 2001,194, the 95-100 page or leaf, its disclosure is incorporated this paper by reference into, has described NF and the expression of NF receptor on melanoma cell.Therefore skin cancer cell, especially melanoma cell can be comprised in the list of NF-receptor-positive cancer cell above.

Because above explanation is relevant with neurological's disease, antibody, polynucleotide and anti--proteic use of NF receptor mutation are more made us expectation (referring to people such as LeSauteur, Nature Biotech, the 1996,14,1120th page) not as micromolecular use.Similar with the proteic model of action of NF, the steady-state adjustment that is used for NF will be expected with the micromolecule agent of the trk receptor of the binding partners anticancer through the control receptor very much.

The present invention is to cause being the basis with the new discovery that non-toxicity mode is regulated NF and kept injury-free we of the normal homeostatic control process of curee like A/B-cis furostan, furan steroid alkene, spirostane or spirostene steroid sapogenines and its ester, ether, ketone and glycosylated form as described in hereinafter described.Therefore, induce the stable state of self regulating of NF and almost be free from side effects for said dose, if said side effect exist also be can be managed and do not hinder using of effective dose.Use micromolecule nontoxic basically, non-peptide to induce the stable state of self regulating; The said discovery that to not have disadvantageous side effect in one or more NF reparation (through the level that increases or reduce) to health status of non-health status is unexpected and astonishing whereby; And significant benefits is provided, like what hereinafter discuss in more detail.

And, we have found that said dose induce one or more NF for example BDNF and GDNF the stable state of self regulating and do not have disadvantageous side effect.It is astonishing obtaining not have disadvantageous side effect more than a kind of NF stable state of self regulating together through a kind of activating agent, and as far as we know, in any micromolecule agent, is unique.Because known neuron need be used for best neuroprotective and nerve repair function usually more than a kind of NF, this discovery according to the present invention provides the disease of NF mediation and the improved in essence processing and the prevention of related conditions.

Also known NF works in tissue comprises the healing of skin, cornea tissue, bone and muscle and helps skin, bone and muscle health usually.Referring to for example Albers, people such as K.M., Neuroscientist 2007,13, the 317-382 page or leaf; Asaumi, K. waits the people, Bone, 26 (6), in June, 2000,625-633 page or leaf; You, people such as L, Investigative Opthalmology Visual Science, October calendar year 2001,42 (11), 2496-2504 page or leaf; Cruise, people such as B.A., Developmental Biology, 271, (2004), 1-10 page or leaf; Jurjus, people such as A., Burns 33 (2007), 892-907; Matsuda, H. waits the people, J.Exp.Med., 187 (3), on February 2nd, 1998,297-306 page or leaf; Menetrey, people such as J, J.Bone Joint Surg (Br), 82-B (1), in January, 2000,131-137 page or leaf; Micera, A. waits the people, Cytokine & Growth Factor reviews, 18, (2007), 245-256 page or leaf; Nithya, M. waits the people, Biochim.Biophys.Acta, 1620, (2003), 25-31 page or leaf; Matsuda, people such as H, J.Exp.Med.1998,187, the 297-306 pages or leaves; People such as Lambiase, Invest.Ophthalmol.Vision Sci, 2000,41,1063-1069 page or leaf.The content of these publications is incorporated this paper by reference into.

Find therefore also can be used to organize as these of basis of the present invention and comprise skin, bone, muscle and the part tissue of eye for example healing and the Ankang of cornea tissue.Therefore; Recovery or normalization and tissue (for example skin, bone, eye and the muscle) healing that the invention still further relates to neuronal function in any infringement or unusual tissue or relevant with any infringement or unusual tissue assisted with general skin, bone and muscle health; Comprise muscle and organize the recovery from take exercise, work or consume that skin is from sun exposure, storm dew, rainwater exposure, cold exposure, aging and corrugated recovery, raising endurance and the tired sensation of minimizing.Not not restrictedly, can comprise the healing of wound and burn by the auxiliary organization healing of the present invention, as hereinafter describe in more detail.

The invention summary

According to a first aspect of the invention; The method of in said curee, inducing the stable state of self regulating of NF through under homeostatic control, regulating curee's neurotrophic factor (NF) with avirulent mode is provided, has comprised one or more agent that are selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines and its ester, ether, ketone and glycosylated form of using effective dose to the curee.Curee's natural NF can be a kind of among BDNF and the GDNF or both.

The method of a first aspect of the present invention makes the inducing with limited and manageable side effect of the stable state of self regulating of NF take place.

In a particularly preferred embodiment of a first aspect of the present invention, inductive stable state is regulated among curee's the natural NF two kinds or more kinds of together, for example BDNF and GDNF.

The agent that is selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines and its ester, ether, ketone and glycosylated form of the method for the stable state of self regulating that is used for inducing said curee through under homeostatic control, regulating curee's natural NF with avirulent mode NF is provided according to a second aspect of the invention.

The agent of use according to a second aspect of the invention makes the inducing with limited and manageable side effect or adverse side effect of the stable state of self regulating of NF take place.

According to a third aspect of the invention we; A kind of compositions is provided, and it comprises the activating agent that is selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines and its ester, ether, ketone and glycosylated form of the method for the stable state of self regulating that is used for inducing said curee through under homeostatic control, regulating curee's natural NF with avirulent mode NF.

Make the inducing of the stable state of self regulating of NF take place according to the compositions of the use of third aspect present invention with limited and manageable side effect or adverse side effect.

According to a forth aspect of the invention, purposes in the medicine of the agent that is selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines and its ester, ether, ketone and glycosylated form induces NF in said curee through under homeostatic control, regulating curee's natural NF with avirulent mode in preparation the stable state of self regulating is provided.

Purposes according to a forth aspect of the invention makes the inducing with limited and manageable side effect or adverse side effect of the stable state of self regulating of NF take place.

The present invention has limited adverse side effect, and especially for example the mistake of NGF is induced, overstimulation or cross and to strengthen relevant side effect, side effect relevant with the effect of receptor (antagonist) agonist and the side effect of being correlated with the enzyme combination with NF.

The present invention can be in human and non-human animal curee with handle disease that NF mediates especially neurological, psychiatry, inflammatory, allergia, immunity and tumprigenicity disease method and and in any infringement or unusual tissue or with any infringement or unusual tissue relevant neuron and the recovery or the normalization of other functions; Comprise when aid in tissue (for example skin, bone, eye and muscle) healing and general skin, bone, eye and muscle health, and the non-Therapeutic Method of being correlated with is united use.

Term used herein " NF mediation " should be with general meaning understanding, comprise neurotrophic factor wherein be understood that to the disease or the patient's condition development, carry out or influence the disease and the patient's condition of the property contributed effect.Therefore, for example, wherein existing evidence relates to the disease or the patient's condition of other activators or the inhibitor of NF receptor, its (antagonist) agonist or NF, and these diseases or the patient's condition will be understood that " the NF mediation " according to the present invention.According to the present invention, these diseases and patient's condition intended response are in the steady-state adjustment of the mankind or non-human animal curee's natural NF.

Therefore the present invention can for example pass through damage with any infringement or unusual organizing; Through ischemia, through aging or (in the example of skin) through wrinkling or unite use through the normal neuron or the recovery of other functions that are exposed in the tissue that sunlight, wind, rainwater, cold or other infringement media damage (no matter cerebral tissue or its hetero-organization for example skin, bone, eye and muscle).The recovery of normal neurons function usually according to the present invention through the stable state of self regulating of inducing NF cause neuranagenesis and improved blood flow and the neuropathy patient's condition or neuron unusually for example the normalization of the inflammation in central nervous system (CNS) or the peripheral nervous system (PNS) obtain.

The present invention can unite use with the auxiliary speed and the quality that especially improves the healing of human and other mammalian skin wounds of wound healing thus.Under this background, " wound " comprises all infringements in any source, for example damages such as incised wound and scratch, knife injury, operation wound, contusion, burn, infections, wound.Chronic and acute wounds all can be handled according to the present invention.

The present invention can unite use with fetal cell, stem cell or other cell therapys and tissue transplantation, especially with the effect of the survival that improves graft materials or therapy or both.Instance comprises the cell therapy of cell function in other organs that improve brain function or health.

Also additionally; The present invention can use in the method for non-treatment so that owing to the benefit of the stable state of self regulating of NF in the tissue promotes or auxiliary these are organized the for example Ankang and the general health of skin, bone, eye and muscle; Promote muscle and the recovery of tissue from taking exercise, working or consume; Promote skin from aging, wrinkling or be exposed to the recovery of sunlight, wind, rainwater, cold or other infringement media, improve the sensation of endurance and muscle endurance (for example in competitiveness or noncompetitive move) and minimizing fatigue.

According to the present invention, said dose can be by system or use partly, because they are found normally good to sending of site of action.Especially, but without limitation, find oral and parenteral (for example (topical) of external) application route is suitable, like what hereinafter discuss in more detail.

The employed statement of this paper " ruscogenin " comprises the derivant of all E and/or F ring opening, and the pseudo-ruscogenin of said ruscogenin and dihydro puppet ruscogenin form for example is possible to the bearing of process of these derivants.In unsaturated (alkene) form of chemical compound, one or more pairs of keys are present in the position that does not influence A/B cis primitive.The glycosylation form of ruscogenin is commonly called Saponin.

Detailed Description Of The Invention

Foreword

The evidence that shows among the application shows that said dose does not combine (referring to embodiment 1) with many receptors and enzyme.

The application has shown and has supported the evidence of activating agent to the effect of inducing NF or NF-receptor.Said evidence (embodiment 2 and 3 vide infra) shows the active expression that strengthens NF and NF-acceptor gene that comprises.As can be seen in embodiment 2, wherein said neuron is healthy (basal medium) relatively.Enhanced gene expression is instantaneous and time-quantum method is indicated the participation of self regulatory mechanism strongly.

In the more unsound situation of embodiment 3, the enhanced gene expression of data show prolong much longer period, show that regulatory mechanism is kept perfectly and the enhanced degree of gene expression depends on the needs of system.

The evidence that shows among the application shows that equally activating agent provides NF the for example stable state (embodiment 4 to 7 and 18 vide infra) of self regulating of BDNF and GDNF especially.Not only a kind of NF for example BDNF or the GDNF normalization of self regulating through non-peptide agent is rare, and two kinds of NF for example BDNF and GDNF together the normalization of self adjusting through non-peptide agent also be unique.Two kinds of NF normalization together shows the combination of the collaborative normalization that causes particularly advantageous BDNF and GDNF.

The evidence that shows among the application shows that equally (referring to embodiment 8) takes place the neurite that activating agent increases in a large amount of CNS and the PNS neuron.Important ground, this neurite is had an effect and is not depended on the existence of exogenous NF.This shows that of the present invention dose effect is that NF induces rather than strengthens.

The evidence that shows among the application shows that equally activating agent activates the endocellular transduction path (referring to embodiment 9) the same with NF.This provides said dose NF to regulate active supporting evidence.

The evidence that shows among the application is same, and to show that many A/B cis activating agents reduce glutamate, Glus inductive to the infringement of cortical neuron and the apoptosis of dopaminergic neuron, yet have roughly similar chemical constitution but the ruscogenin (diosgenin) that do not have an A/B cis primitive is inactive (referring to embodiment 10 and 11).

The evidence that shows among the application is same to show that said dose of neuron that reverses in many neurons damages, and promptly they are healing nerve or neuranagenesis (referring to embodiment 12).

The same demonstration of the evidence that shows among the application is (referring to the embodiment 13 and 14) that orally-ingestible is used for said dose.Said embodiment shows that said dose Orally administered improve the recovery of function of nervous system in the mouse model of nerve injury after motor neuron disease or the wound.

The evidence that shows among the application is same to show that said dose is reduced the anxiety in the old rat and repair cognitive (referring to embodiment 15).

The evidence that shows among the application is same to be shown that Orally administered agent is delivered to many bodily tissues (referring to embodiment 16) and is nontoxic (referring to embodiment 17) in effective dose.

Said dose of parkinson (referring to embodiment 18) that reduces in the macaque of the same demonstration of the evidence that shows among the application.

Evidence demonstration NF that shows among the application or said dose activity of NF-receptor (NFr) mediation do not comprise the direct binding interactions with many receptors and enzyme.For example; Exciting direct of the direct bonded excitement of active and many important receptors, antagonism or non-(antagonism) combines irrelevant; Said important receptor comprises hormone receptor for example estrogen, progesterone, testosterone and serotonin receptor; Nicotinic receptor; M-ChR; Adrenoreceptor, narcotic receptors be cannabinoid and opiate receptor for example, and glutamate receptor is NMDA, AMPA and kainite receptor and retinoic acid receptors retinoid X receptor for example for example.As a result of, the physiological effect of activating agent does not rely on the receptor of the processing discovery of known neurological of being used for and psychiatric disorders before and many side effect of the side effect that enzyme mediates.For example; When using said dose; The problem that many processing before of the neurological and the psychiatry patient's condition are found reduces in a large number; Wherein addiction property and dependency; The habit-forming personality type, have receptor or enzyme side effect before processing and existing break addiction or dependent processing each all can show dealing with improperly of neurological or psychiatric disorders.

The prior art processes and displays of psychiatric disorders goes out the favourable psychiatry effect of much longer time-quantum method, although they bring into play the effect of their biochemistry patterns immediately.Effect of the present invention is much instant, is provided under the homeostatic control evidence with the adjusting of the NF of atoxic mode.Therefore the present invention handles different with the known micromolecule (non-peptide) that is used for the P&N disease.

In the test of embodiment said dose dose response overview is the maximum of plateau after showing, this is the characteristic (referring to Fig. 1) of self regulatory mechanism.

Employed one or more activating agents can be used neurotrophic factor in external source not and for example use under GDNF or the BDNF among the present invention.

New purposes related to the present invention

Therefore the present invention makes the new purposes of the activating agent of subject to confirmation become possibility, for example the new purposes aspect following: (i) pending disease is arranged, the type of pending individuality is (ii) arranged, combined treatment to be used and situation about (iv) using safely are (iii) arranged.

With regard to (i); For example; Via pharmaceutical preparation and functional food; Activating agent is confirmable handling following purposes in a series of now; Like what discuss in further detail hereinafter: the neurological; Psychiatry; Inflammatory; Allergia; Immunity and tumprigenicity disease and the patient's condition and personality and behavior characteristics and realization neuron; Regeneration and normalization to neuronic blood flow; Impaired tissue (skin for example; Bone; Eye or muscular tissue) regrowth and healing; The inside and outside tissue of brain (skin for example; Bone; Eye or muscular tissue) general health and Ankang; Muscle and tissue are from taking exercise; Work and the recovery that consumes; Improve endurance and reduce tired sensation, regenerate normal neuronal function and normal neuroid.This purposes comprises neurological or psychological function or the individual general health and safe and comfortable non-therapeutic use of improvement at the individuality of crowd in normal range; Improve skin; Bone; Eye; The non-therapeutic use of muscle and other tissue health; For example promote skin from aging; Wrinkling or be exposed to sunlight; Wind; Rainwater; Effect cold or other infringement media is restored; And the non-therapeutic use that provides for other healthy with safe and comfortable aspects; Comprise muscle and organize from taking exercise; Work or consumption are restored; Improve endurance and reduce tired sensation, and term " disease "; " patient's condition " and " speciality (trait) " should correspondingly be understood.

With regard to (ii), of the present invention dose via NF self regulate stable state rather than to the adjusting of many receptors or enzyme or combine to come that acting discovery allows to remain to be processed to the responsive patient of adverse side effect away from some enzyme inhibitor things or receptor agonism medicine.For example, some dementias (Alzheimer) patient can not tolerate cholinesterase inhibitor.Some parkinsonians can not tolerate levodopa and will suffer hardships in the side effect that comprises that the dyskinesia or neuropsychiatry problem are for example taken a risk.

With regard to (iii), for example, the said dose of purposes with the combination of other adjuvant of the individuality that is used for special disease, the patient's condition and speciality or special type is confirmable now, like what hereinafter introduce more in detail.Many confirming in this combination inferred before at most.This purposes comprises that raising is at the neurological of the individuality of crowd in normal range or the non-therapeutic use of psychological function; Improve skin; Bone; Eye; The non-therapeutic use of muscle and other tissue health; For example promote skin from aging; Wrinkling or be exposed to sunlight; Wind; Rainwater; Effect cold or other infringement media is restored; And the non-therapeutic use that provides for other healthy with safe and comfortable aspects; Comprise muscle and organize from taking exercise; Work or consumption are restored; Improve endurance and reduce tired sensation, and term " disease "; " patient's condition " and " speciality " should correspondingly be understood.

With regard to (iv), for example, the new range of situation about using outside clinical and the pharmacy situation is confirmable now, like what hereinafter introduce more in detail.

New purposes (i)-Xin handles

The present invention can be used to following method (a) and handle or prevent neurological, psychiatry, inflammatory, allergia, immunity and tumprigenicity disease; (b) regeneration and/or normalization neuron and extremely neuronic blood flow; Comprise regenerating nerve meta function or neuroid; (c) regrowth of Sun Hai tissue and healing; (d) muscle and tissue from take exercise, work or the recovery that consumes, (e) improve endurance and reduce tired sensation or (f) in its mankind or non-human mammal of needs, handle or prevention Deviant Behavior or personality trait.Said neurological, psychiatry, inflammatory, allergia, immunity and tumprigenicity disease can be in the prior art mentioned above disclosed those or can be different with those diseases.For example, neurological's method can be lonely syndrome, depression and schizophrenia, maybe can be the disease that is different from these.Be used for neuron and to the regeneration or the normalization of neuronic blood flow; The regrowth of injured tissue and healing; The method of neuronal function or neuroid is rebuild after comprising for example neural wound; The operation of tissue transplantation and nerve (for example; The reatlachment that is used for extremity and finger) back rebuilds; Auxiliary from apoplexy, instantaneous cerebral ischemia attack (TIA) or the recovery of other ischemias; For example auxiliary nervous function and to the recovery of the blood flow of ischemic tissue, the healing of auxiliary wound, bone and muscle and handle the neuropathy and any inflammatory patient's condition relevant with CNS or PNS.

The neuroprotective of considering activating agent acts on neural repair (neuranagenesis); The present invention can unite use with fetal cell, stem cell or other cell therapys, and said therapy for example is used for neurological and psychiatric disorders or is used for the recovery or the normalization of undermined or unusual tissue or function.Instance comprises the cell therapy of handling encephalopathy disease.Can improve the effectiveness of cell therapy according to the use of activating agent of the present invention, for example through increasing the survival rate of the cell of transplanting, through increase handle in effectiveness or its combination of cell of survival.

The present invention can be used to suffer from or be prone to suffer from the method for processing of the mankind or this disease among the non-human animal of the disease relevant with the unconventionality expression of one or more NF or NFr.Disease can be in the prior art mentioned above disclosed those or can be different with those diseases.For example, neurological's method can be lonely syndrome, depression and schizophrenia, maybe can be the disease that is different from these.These diseases except neurological or psychiatric disorders or Deviant Behavior or personality trait comprise sleep deprivation for example with stress effect, inflammatory disease, allergy, immune disorders and NF cancers mediated.

As above mentioned, evidence in this application shows that employed activating agent can be with level normalization or the enhancing simultaneously of BDNF in the brain and GDNF among the present invention.The method of one or both while normalizations in these NF levels in the mankind of the brain level that the present invention therefore can be used to suffer from one or both unusual among BDNF and the GDNF or to reduce or non-human animal's the brain.

The present invention provides the for example method of the stable state of self regulating of BDNF and GDNF of NF of inducing.Said method makes the inducing with limited and manageable side effect of the stable state of self regulating of NF take place.The application comprises the evidence that this induces the existence that does not require peptide NF or NFr.Therefore, opposite with known dose, the present invention has avoided the needs of using altogether to peptide NF or NFr and one or more non-peptide activating agents of the present invention.

In the above described new purposes each can be in aspect of the present invention each use.

The present invention so as to the method for generation effect can be treatment or non-treatment and compositions can be medicine or non-pharmaceutical composition, like what hereinafter describe in detail more.Although other application route are provided, activating agent is preferably by Orally administered, like what hereinafter describe in detail more.

New purposes (ii)-newtype of accessible individuality

New discovery as the present invention basis shows that activating agent can be used to handle; At least in certain hour; Can be natural cross and express or the individuality of one or more NF of unconventionality expression or NFr (for example BDNF and/or GDNF); For example sleep deprivation or stress the people, be incompatible and handle these individualities through NF simulation or stimulant before.

The disease that the present invention can be used to handle or prevent to suffer from or easy trouble and minimizing or unusual NF or NFr level are relevant or the mankind or this disease among the non-human animal or the method for the patient's condition of the patient's condition, the said mankind or animal are that natural the mistake expressed or the NF or the responsive individuality of NFr of unconventionality expression one or more other.

New discovery as the present invention basis shows that also activating agent can be used to handle the responsive individuality of psychiatry side effect to NF-simulation or stimulating drug; These side effect are psychiatry, mood, anxiety or other personalities or behavior symptom normally, is incompatible through NF simulation or stimulant to the processing of said individuality before.

Of the present invention dose via the stable state of self regulating of NF rather than to the adjusting of many receptors or enzyme or combine to come that acting discovery allows to remain to be processed to the responsive patient of adverse side effect away from some enzyme inhibitor things or receptor agonism agent medicine.For example, some dementias (Alzheimer) patient can not tolerate cholinesterase inhibitor.Some parkinsonians can not tolerate levodopa and will suffer hardships in the side effect that comprises that the dyskinesia or neuropsychiatry problem are for example taken a risk.

The present invention can be used to handle or prevent and reduce or the disease that unusual NF or NFr level are relevant or the mankind or this disease among the non-human animal or the method for the patient's condition of the patient's condition with suffering from or be prone to suffer from, and the said mankind or animal are the individualities to the psychiatry of NF-simulation or stimulant or other side effect sensitivity.

Comprise for example neurological, psychiatry, inflammatory, allergia, immunity and tumprigenicity disease or Deviant Behavior or personality trait with the minimizing or the unusual NF or the relevant disease or the patient's condition of NFr level, for example hereinafter in greater detail those.In addition, these diseases and the patient's condition comprise hereinafter describes skin, muscle, eye and bone disorders and the patient's condition, comprises the patient's condition of the fatigue of the patient's condition relevant with health with the Ankang of tissue and muscle or its hetero-organization.

Show also that as the new discovery on the present invention basis a large amount of hormones and other receptors are not had (antagonism) excitement or binding ability and do not have the activating agent of enzyme binding ability can be used to handle to the receptor of medicine or the responsive individuality of side effect of enzyme mediation to many enzymes.These individualities can for example comprise having addiction property or dependent individuality, and said addiction property or dependency can be through being subjected to said addiction property or relying on (antagonism) agonism aggravation of sex receptor; Handle or self processing procedure in remain to be given up addiction property or dependent individuality, wherein can regress through (antagonism) agonism that is subjected to said addiction property or relies on sex receptor because same reason is given up process; Having addiction property or dependent personality type for example has (antagonism) excitement on the receptor or combination or enzyme is combined some the responsive especially receptor or the individuality of metabolic process.And; Can occur at those individualities of processing of interferential other clinical patient's condition that stand to be used for receiving the effect of these receptors or enzyme mediation the sensitivity of receptor or enzyme mediation side effect, for example stand the individuality of HORMONE TREATMENT (for example the HORMONE TREATMENT on the oncology, growth hormone are handled, thyroxin is handled, estrogen alternative medicine (HRT) or sex reformation therapy).

Therefore the present invention can be used to handle or prevent the disease relevant with NF that suffers from or be prone to suffer from and reduce or NFr level or the mankind or this disease among the non-human animal or the method for the patient's condition of the patient's condition, and the said mankind or animal are the responsive individualities of side effect to the receptor of medicine or enzyme mediation.

With individuality the relevant receptor of the sensitivity of the side effect of receptor (or binding site) mediation or binding site are comprised any or multiple in the following receptor: adensonine A ₁Receptor; AdensonineA _2AReceptor; Adensonine A ₃Receptor; Non-selective adrenergic α 1 receptor comprises adrenergic α _1A, adrenergic α _1BOr adrenergic α _1DReceptor; Non-selective adrenergic alpha-2 receptor comprises adrenergic α _2AOr adrenergic α _2CReceptor; Non-selective adrenergic comprises alpha 1 beta-adrenergic ₁, alpha 1 beta-adrenergic ₂Or alpha 1 beta-adrenergic ₃Receptor; Adrenomedullin AM ₁Receptor; Adrenomedullin AM ₂Receptor; Aldosterone receptor; The c5a anaphylatoxin receptor; Androgen (testosterone) receptor AR; Angiotensin AT ₁Receptor; Angiotensin AT ₂Receptor; Apelin (APJ) receptor; The atrial natriuretic factor receptor; Bombesin BB1 receptor; Bombesin BB2 receptor; Bombesin BB3 receptor; Kallidin I B ₁Receptor; Kallidin I B ₂Receptor; Calcitonin receptor; Calcitonin-gene-related peptide (CGRP ₁) receptor; Benzodiazepine L-type calcium channel; Dihydropyridine L-type calcium channel; Phenylalkylamine L-type calcium channel; Calcium channel N-type; Cannabinoid CB ₁Receptor; Cannabinoid CB ₂Receptor; Chemotactic factor CCR1 receptor; Chemotactic factor CCR2B receptor; Chemotactic factor CCR4 receptor; Chemokine ccr 5 receptor; Chemotactic factor CXCR1 receptor; Chemotactic factor CXCR1 (IL-8RB) receptor; Cholecystokinin CCK ₁(CCK _A) receptor; Cholecystokinin CCK ₂(CCK _B) receptor; The Colchicine receptor; Corticotropin-releasing factor (CRF ₁) receptor; Dopamine D ₁Receptor; Dopamine D _2SReceptor; Dopamine D ₃Receptor; Dopamine D ₄₂Receptor; Dopamine D ₅Receptor; Endothelin ET _AReceptor; Endothelin ET _BReceptor; Epidermal growth factor (EGF) receptor; Erythropoietin EPOR receptor; Estrogen receptor; Estrogen (ER α) receptor; Estrogen (ER β) receptor; G protein coupled receptor GPR103; G protein coupled receptor GPR8; GABA _AReceptor; TBOB chloride channel GABA _AReceptor; Maincenter flunitrazepam GABA _AReceptor; Maincenter 5-aminomethyl-3-hydroxyisoxazole GABA _AReceptor; GABA _B1AReceptor; GABA _B1BReceptor; The gabapentin receptor; Galanin GAL1 receptor; Galanin GAL2 receptor; The glucocorticoid receptor; Glutamate receptor; The AMPA glutamate receptor; The kainate glutamate receptor; Exciting NMDA glutamate receptor; Glycine NMDA glutamate receptor; Phencyclidine NMDA glutamate receptor; Polyamines NMDA glutamate receptor; Growth hormone secretagogues (GHS, Ghrelin) receptor; Histamine H ₁Receptor; Histamine H ₂Receptor; Histamine H ₃Receptor; Histamine H ₄Receptor; Maincenter imidazoline I ₂Receptor; Inositoltriphosphoric acid IP ₃Receptor; Insulin receptor INSR; Interleukin I L-1 receptor; Interleukin I L-2 receptor; Interleukin I L-6 receptor; The Leptin receptor; BLT leukotriene (LTB ₄) receptor; Cysteinyl leukotriene CysLT ₁Receptor; Cysteinyl leukotriene CysLT ₂Receptor; Melanocortin MC ₁Receptor; Melanocortin MC ₃Receptor; Melanocortin MC ₄Receptor; Melanocortin MC ₅Receptor; Melatonin MT ₁Receptor; Melatonin MT ₂Receptor; The motilin receptor; Muscarinic M ₁Receptor; Muscarinic M ₂Receptor; Muscarinic M ₃Receptor; Muscarinic M ₄Receptor; Muscarinic M ₅Receptor; N-formyl peptide receptor FPR1; N-formyl peptide receptor appearance FPRL1 receptor; Neuromedin U MNU ₁Receptor; Neuromedin U MNU ₂Receptor; Neuropeptide tyrosine Y ₁Receptor; Neuropeptide tyrosine Y ₂Receptor; Neurotensin NT ₁Receptor; NAChR; Nicotinic acetycholine α 1, the bungarotoxin receptor; Nicotinic acetycholine α 7, the bungarotoxin receptor; Opium δ (OP1, DOP) receptor; Opium κ (OP2, KOP) receptor; Opium μ (OP3, MOP) receptor; Nociceptin ORL ₁Receptor; Buddhist ripple ester receptor; Platelet activating factor (PAF) receptor; Platelet-derived somatomedin (PDGF) receptor; Potassium channel [K _A]; Potassium channel [K _ATP]; Potassium channel [SK _CA]; Potassium channel HERG; Progesterone receptor; Progesterone PR-B receptor; Prostanoid CRTH2 receptor; Prostanoid DP receptor; Prostanoid EP ₂Receptor; Prostanoid EP ₄Receptor; The prostanoid thromboxane A ₂(TP) receptor; Purine can P _2xReceptor; Purine can P _2YReceptor; Tetanus appearance X receptor RXR α; The rolipram receptor; Li Luoding (ryandine) RyR3 receptor; Serotonin serotonine 5-HT1 receptor; Serotonine 5-HT _1AReceptor; Serotonine 5-HT _1BReceptor; Serotonine 5-HT _2BReceptor; Serotonine 5-HT _2CReceptor; Serotonine 5-HT ₃Receptor; Serotonine 5-HT ₄Receptor; Serotonine 5-HT _5AReceptor; Serotonine 5-HT ₆Receptor; σ (sigma) σ 1 receptor; σ (sigma) σ 2 receptors; Site 2 sodium channel receptors; Somat (somastatin) sst1 receptor; Somat sst2 receptor; Somat sst3 receptor; Somat sst4 receptor; Somat sst5 receptor; Tachykinin NK-1 ₁Receptor; Tachykinin NK-1 ₂Receptor; Tachykinin NK-1 ₃Receptor; Testosterone receptor; Thyroid Hormone Receptors; Throtropin releasing hormone (TRH) receptor; Transforming growth factor-beta (TGF-β) receptor; The vidarabine carrier; The choline carrier; Dopamine carrier (DAT); The GABA carrier; The monoamine carrier; Norepinephrine carrier (NET); Serotonine carrier (SERT); Non-selective tumor necrosis factor (TNF) receptor; Urotensin II receptor; Novel vanilloid receptor; VEGF (VEGF) receptor; Vasoactive intestinal peptide VIP ₁Receptor; Vassopressin V _1AReceptor; Vassopressin V _1BReceptor; Vassopressin V ₂Receptor and vitamin D ₃Receptor.

With individuality the relevant enzyme of the sensitivity of side effect is comprised in the following enzyme one or more: acetylcholinesterase; The acetyl-CoA synzyme; Choline acetyltransterase; Albumen serine/threonine kinase AKT1 (PRKBA); Albumen serine/threonine kinase AKT3 (PRKBG); Albumen serine/threonine kinase CAMK2D (KCC2D); Albumen serine/threonine kinase MAP2K1 (MEK1); Protein serine/threonine kinase MAPK1 (ERK2); Albumen serine/threonine kinase MAPK11 (p38 β); Albumen serine/threonine kinase MAPK12 (p38 γ); Albumen serine/threonine kinase MAPK13 (p38 δ); Albumen serine/threonine kinase MAPK3 (ERK1); Serine/threonine protein kitase MAPK8 (JNK1); Non-selective albumen serine/threonine kinase PKC; Protein tyrosine kinase NTRK1 (trkA); Protein tyrosine kinase NTRK2 (trkB); Protein tyrosine kinase SRC; Aldose reductase; ABTS atomic group free radical scavenging enzyme; DPPH atomic group free radical scavenging enzyme; SOD intends thing free radical scavenging enzyme; With UDP glucuronide transferase UGT1A1.

Therefore, the present invention makes the micromolecule therapeutic agent possibly be used for the active side effect that these individual and acomia receptors of being conigenous activating agent and enzyme mediate first or has the risk of this side effect of generation of remarkable minimizing at least.

Some A/B cis spirostane ruscogenin and Saponin have been disclosed among WO-A-99/48507, WO-A-99/48482, WO-A-01/23406, WO-A-01/23407, WO-A-01/23408 and the WO-A-01/49703 before estrogen, androgen, progesterone, glucocorticoid and the testosterone receptor non-activity.Although shown muscarinic receptor number and the synthetic evidence that increases, do not show that identical agent is to muscarinic receptor non-activity/do not combine.The evidence of the evidence of the muscarine and the number normalization of alpha 1 beta-adrenergic 2 receptors-not relevant-show in WO-A-02/079221 and WO-A-03/082893 with dose dependent or activity/combination.

The number of nicotine receptor has been disclosed WO-A-99/16786 (EP-A-1024146 before via the evidence that some A/B cis furostan Saponin, zhimusaponin BII dose dependent increase; US-A-6593301) in.The said dose of activity to these receptors/bonded degree obviously do not measured.Existing other evidence shows that the effect of reporting in the prior art comes from the synthetic of NF and/or their receptor or discharges the increase of adjusted and/or the minimizing on the degradation speed.

The present invention can be used to handle or need prevent its mankind or the neurodegeneration among the non-human animal and do not induce to relate to and above walk to receptor listed in the 18th page of the 29th row and one or more receptor or the side effect of enzyme mediation in the enzyme for the 16th page the 19th.

Said method can be the compositions that compositions therapeutic or non-therapeutic and said can be medicine or non-medicine, and is as described in greater detail below.For example, the purposes of non-therapeutic can be neurological or the psychological function that improves at the individuality of crowd in normal range.Term " disease ", " patient's condition " and " speciality " should correspondingly be understood.Although other application route are provided, activating agent is preferably by Orally administered, like what hereinafter describe in detail more.

New purposes (iii)-the new combination of agent

Activating agent among the present invention can be learned the activating agent combination with known or doubtful other biological for the abnormal level (promptly unusual low or unusual high level) that possibly cause NF or NFr among the curee and use, or can be on the basis of prevention with unknown or do not suspect or one or more other biologicals activating agents of not being detected the probability that causes this abnormal level use.These other biologic activity agent comprises for example pharmaceutical preparation of activity chemistry agent, Profilin or polynucleotide (for example antibody, antibody fragment such as F (ab) or F (ab) ₂Fragment, siRNA or antisense DNA) specific-binding agent and active mass stem cell for example.

With this method, can be used to resist any detrimental effect that said other biological is learned activating agent according to of the present invention dose.

The present invention can be used for being applied to the mankind or non-human animal curee to handle or prevention patient's a certain illness or the composition or the composition cover group (set) (group of collocation) of the patient's condition are used; Said composition or cover group comprise first kind of bioactivator of the potentiality that are used for handling or prevent the said illness or the patient's condition and have the abnormal level that causes curee NF or NFr; Resist any this unusual NF that the curee induces or the activating agent of the present invention of NFr level with the mode that is used for regulating with self; Unusual NF or NFr level resisted described in the curee thus, preferably tends to normal N F or NFr level.

New purposes (iv)-new situation used

The present invention can be used to wherein to use or the close clinic control non-availability or infeasible situation of treatment sequence in.

The properties of combination of the time lengthening of the patience of the processing of self regulating of excessive administration and response is beneficial to controlling using of activating agent under the relative inadequate situation, and for example self uses or non-therapeutic is used.The program that is used for according to processing method of self regulating of the present invention will be to handle wider patience than corresponding prior art effectively.

Use in the method for the present invention any therefore can be in the situation of the clinic control of not using program, especially self use or situation that non-therapeutic is used in use.

Any aspect of the present invention can with other aspects of the present invention in any one or a plurality ofly carry out simultaneously or use, and any instance that illustrates for one aspect of the present invention or preferably will be applied to any other aspect of the present invention identically.

" handle or prevention "

The employed statement of this paper " is handled or prevention " and similar terms is meant the health care that is intended to remove or avoid disease or alleviates the form of ownership of its symptom; Comprise preventative, healing property and the property alleviated, as any in the test that can get judge according to putting into practice according to popular medical science and psychiatry.Be intended to reasonable expectation obtain particular result but the intervention that always do not realize said result be included in the statement " handle or prevent ".The intervention that successfully slows down or stop the process of disease is included in during statement " handles or prevention ".

Some neurological, psychiatry, inflammatory, allergia and immune disorders are counted as " spectrum (spectrum) " patient's condition, wherein individually can show many some or all of in maybe symptoms, perhaps can only show the said disease of slight form.And many neurological, psychiatry, inflammatory, allergia, immunity and the tumprigenicity patient's condition are progressive, start from slight relatively abnormal symptom and proceed to more serious abnormal symptom.The present invention includes the processing and the prevention of neurological, psychiatry, inflammatory, allergia, immunity and the tumprigenicity patient's condition of all NF mediations, and no matter the said patient's condition is in which kind of type and stage.

" be prone to suffer from "

The employed statement of this paper " be prone to suffer from " refers in particular to the individuality that is in the risk higher than the normal risk that develops medical science, health, Ankang or psychiatric disorders or personality changes with similar terms, is used for like use that the known risks and assumptions of individuality or disease assesses.These individualities can, for example, be classified as remarkable risk with development one or more particular disorder or personality changes, reach and will this individuality opened medicine and/or give the degree of special diet, life style or similar recommendation.

Toxicity and side effect

Have limited and manageable side effect and in use be avirulence or essentially no toxic according to of the present invention dose.

Under the background of pharmacy (comprising the veterinary) purposes; The physiology that this hint is said dose is acceptable; Consequently; In the scope of good medical science and veterinary's judgement; Said dose is suitable for using with the effective dose side effect that no abnormal toxicity, stimulation, allergia respond, do not expect with the cells contacting of the mankind, mammal and other animals; And generable these detrimental effects are considered to over-drastic maybe can not be managed through aid in treatment, can not be equivalent to rational benefit/risk ratio.

For example functional cosmetics and dermatological or other contact skin or eye contact under the background of goods at functional food especially grain, food supplement (comprising dietary supplements), beverage and beverage fill-in and external preparation; The corresponding assessment of this hint benefit/risk and side effect is suitable for use in the safety and the toxicity criterion of particular composition or goods and its special-purpose that is provided.

" non-therapeutic method "

The non-therapeutic purposes is usually by there not being under the medical supervision human subject that the selectivity self of physiologically active agent in the compositions is used, and is normally oral, characterize.Usually; The benefit of resultant expection is and the following patient's condition or relevant Ankang or the general health benefit of the patient's condition awared; The said patient's condition is (i) informal diagnosis; Therefore (ii) not diagnosable according to clinical practice, or (iii) at healthy population in normal range and be not considered to disease.

The non-therapeutic purposes also can be got involved by the stage medical science in curee's purchase or acquisition compositions or auxiliary the existence characterizes.

Also additionally, the non-therapeutic purposes can be characterized by the supplier's who does not have compositions medical requirement, so that self uses and be not to be ordered about by the special intention of handling the disease diagnosed.

For example, can suitably can be comprised, for example by neurological's function of non-therapeutic influence; Cognitive (comprising thinking, reasoning, memory, memory, the imagination and study); Attention and concern are particularly towards slighter end of patient's condition scale and slight Deviant Behavior or personality trait.The psychological function of can be suitably being handled by non-therapeutic can comprise for example behavior of men, emotion, personality and social functions, such as sexual behaviour, sexual dysfunction, sorrow, anxiety, depression, poikilothymia, gloomy, teenager emotion, the sleep pattern of interruption, lively dream (vivid dreaming), nightmare and sleep-walking.

Except the accessible neurological of non-therapeutic method according to the present invention and psychological function that above provide, but, corelation behaviour or thinking can not be considered to non-therapeutic processing according to the present invention according to the slight form of not diagnosable neurological of clinical practice and psychiatric disorders because causing individual serious distress or he or she daily function not have to interrupt yet.

The slight form of inflammatory, allergia and immune disorders or unknown cause or the inflammatory, allergia and the immune disorders that do not have to accept formal diagnosis owing to other reasons also can be considered to the patient's condition that but non-therapeutic is handled according to the present invention.

But benign tumor sexually transmitted disease (STD) disease or unknown cause or also can be considered to non-therapeutic processing according to the present invention because other reasons not have to accept the tumprigenicity disease of formal diagnosis.

" normalization "

Employed statement of this paper " normalization " and similar terms (for example " stable state ") refer in particular to the physiological regulation towards the situation that is characterized as general normal health.Best normal condition can come example by the healthy young adult mankind or non-human animal's situation.

In fact whether " normalization " comprises the process of regulating towards normal condition thus, no matter to reach to be characterized as being normal situation.

Neurological's disease

Employed statement of this paper " neurological's disease " and similar terms comprise for example neurodegeneration (comprising the neurodegeneration of the symptom with impaired cognition and the neurodegeneration that does not have the symptom of impaired cognition), nervimuscular degeneration and motion sense organ neurodegeneration.

The instance of neurological's disease that the present invention pays close attention to includes but not limited to: dementia; The cognitive impairment that age is relevant; Alzheimer; The alzheimer disease of Alzheimer type (SDAT); Dementia with Lewy body; Vascular dementia; Parkinson; Postencephalitic parkinsonism; Parkinson with the reason except that postencephalitic and parkinson; Muscular dystrophy comprises facioscapulohumeral muscular dystrophy (FSH); Duchenne muscular dystrophy; Duchenne muscular dystrophy and Bu Lusishi muscular dystrophy; Fuchs; Myotonic dystrophy; Cerneal dystrophy; Reflex sympathetic dystrophy syndrome (RSDSA); The neural blood vessel malnutrition; Myasthenia gravis; The Eton, Lambert is sick; Huntington Chorea; Motor neuron disease comprises amyotrophic lateral sclerosis (ALS); Baby's spinal cord amyotrophy; Multiple sclerosis; Postural hypotension; Pain; Neuralgia; Traumatic neurodegeneration is for example after apoplexy or after unexpected (for example traumatic head or brain injury or spinal cord injury); Crust Teng Shi is sick; Cockayne syndrome; Mongolism; The degeneration of cortex ganglion basal; Multiple system atrophy; Brain atrophy; Olivopontocerebellar atrophy; Dentato rubral atrophy; Pallidum subthalamic nuclei atrophy (pallidoluysian atrophy); Spinobulbar atrophy; Optic neuritis; Sclerosing panencephalitis (SSPE); Attention deficit disorder; Virus back encephalitis (post-viral encephalitis); Post poliomyelitis syndrome; The Fa Er Cotard, Joubert syndrome, guillain-Barre syndrome; Agyria; Moyamoya, the neuron obstacle of dividing a word with a hyphen at the end of a line, lonely syndrome; Poly glumine is sick; Niemann-Pick disease, progressive multifocal leukoencephalopathy, pseudotumor cerebri; Refsum; The Ze Weige syndrome, supranuclear paralysis, family ataxia; 2 type spinocebellar ataxias; The Rhett syndrome, shy-Drager syndrome, tuberous sclerosis; Creutzfeldt jakob disease; Chronic fatigue syndrome, neuropathy comprise that hereditary neuropathy becomes; Diabetic neuropathy and mitosis neuropathy (mitotic neuropathy) comprise Creutzfeldt-Jakob disease (CJD) based on the neurodegeneration of Protein virus; Anomaly CJD; New anomaly CJD; Mad cow disease (BSE); GSS; FFI; Kuru disease and Alper syndrome; Yue Sefushi is sick; Acute disseminated encephalomyelitis, arachnoiditis, central nervous system's vascular lesion; The forfeiture of limbs function of nervous system; Charcot Marie Tooth, it is sick to restrain cured Bi Shi, leukodystrophy; Be prone to suffer from heart failure; Asthma, epilepsy, auditory nerve degeneration; Degeneration of macula, the optic nerve that retinitis pigmentosa and glaucoma cause is degenerated.

Psychiatric disorders

Statement " psychiatric disorders " comprises everyone mattoid disease, and it is influential to personality and behavior, and especially the ability of getting along with the mankind's thinking, sensation, emotion with other people is relevant.Therefore; Have some overlapping between " neurological " and " psychiatric " disease, and in the present invention when remain through the present invention handle or " psychiatric disorders " of prevention especially true will be directly or indirectly relevant the time with the potential neurological deficit that directly or indirectly influenced by NF or NFr.

Generally speaking, mental illness is not diagnosed as " psychiatric disorders ", removes irrelevant behavior or thinking and causes individual serious distress or he or she daily function to be interrupted.Therefore, diagnosable disease and its processing similar but more not serious or that interrupt have boundary (vide infra) between should being considered to the physiologic function of non-therapeutic.

The instance of the psychiatric disorders that the present invention pays close attention to includes but not limited to: anxiety disorder (acute stress disorder for example; Panic disorder; Agoraphobia; Social phobia; Specific phobia disease; Obsessive compulsive disorder; Posttraumatic stress disorder; Body dysmorphic disorder and generalization anxiety disorder); Property anxiety disorder (vulvismus for example; Male erectile dysfunction; Male orgasmic disorder and female orgasmic disorder); The childhood period disease (hyperkinetic syndrome (ADHD) for example; A Si Burger syndrome; Autism; Conduct disorder; Oppositional defiant disorder; Separation anxiety sexual disorders and Tu Leiteshi disease); Eating disorders (for example nervous anorexia and bulimia nervosa); The dysthymic disorder is (for example depressed; Serious depressibility obstacle; Bipolar disorder (manic depression); Seasonal affective disorder (SAD); Cyclothymic disorder and dysthymic disorder); Sleep disorder; Cognitive psychiatric disorders (delirium for example; Amnesia); Personality disorder (paranoid personality disorder for example; Schizoid personality disorder; The schizotypal personality disorder; Antisocial personality disorder; Borderline personality disorder; The histrionic personality disorder; Narcissistic personality disorder; Avoidant personality disorder; Dependent personality disorder and obsessive-compulsive personality disorder), mental disorder (schizophrenia for example; Delusional disorder; Brief psychotic disorder; Schizophreniform disorder; Schizoaffective disorder and shared psychotic disorder) and the relevant disease (substance-related disorder) of material (alcohol dependence for example; The dependence of amphetamine-type stimulants; Cannabis relies on; Cocaine relies on; Hallucinogen dependence; Inhalant dependence; Nicotine dependence; Opioid dependence; Phencyclidine dependence and tranquilizer rely on).

Inflammatory and allergic conditions

The instance of accessible inflammatory and allergic conditions comprises cough according to the present invention, and pruritus (referring to Johansson, people such as O, Arch.Dermatol.Res.; 2002,293, the 614-619 page or leaf), food intolerance; Psoriasis, croup, irritable bowel syndrome, tinnitus; Prunus mume (sieb.) sieb.et zucc. Ni Ershi disease, the inductive ulcer of the ulcer of stress-induced or aspirin, allergic rhinitis, allergic dermatitis; Conjunctivitis, inflammation, inflammatory bowel, ileitis; Pancreatitis, cholecystitis, anallergic rhinitis; Esophagitis, osteoarthritis, rheumatoid arthritis; Hay Fever is to the allergic effect reaction of room demodicid mite (house mite), to the allergic effect reaction of house pet; Huntington Chorea, acute inflammation pain, Encelialgia; Tooth pain and headache, inflammatory hyperpathia, sense of touch hyperpathia is (referring to for example Ma; People such as QP, Neuroreport 1997,8; The 807-810 page or leaf), allergic skin reaction, the reaction of allergia eye; Asthma (referring to Bonini, people such as S, Proc.Natl.Acad.Sci.USA; 1996,93, the 10955-10960 page or leaf; Braun, people such as A, Am.J.Respiratory Cell Mol.Biol., 1999,21,537-546 page or leaf), atherosclerosis, arthritis, chronic ulcer (for example relevant chronic angionoma) and eczema with rheumatoid arthritis.

Relevant non-therapeutic processing according to the present invention comprises keeps eupnea (maintaining normal breathing); Alleviate throat pain and cough; Normal stool is kept in help, alleviates stomach discomfort, helps to restore from flu and influenza; As decongestant; Alleviate headache, alleviate myalgia, relax slight painful and pain; Alleviation to toothache is provided, provides the alleviation of oral cavity or gastric ulcer and the joint of keeping health.

Immune disorders

The instance of the immune disorders of accessible NF mediation comprises the disease that the normalization of effect that can be through immune cell function listed in the table 2 (the 8th page) of NF to people's publications such as Vega of above quoting is handled according to the present invention.These diseases comprise for example AIDS (wherein the normalization of NF will improve curee's immunocompetence) of the immunodeficiency patient's condition; Immunity is the patient's condition (wherein the normalization of NF is more special to external dose with the skeptophylaxis system) of sexually transmitted disease (STD) condition (wherein the normalization of NF will be reduced curee's immune system) and impaired immunologic opsonin excessively, for example autoimmune disease such as systemic lupus erythematosus (sle) (SLE).

The tumprigenicity disease

The instance of the malignant neoplastic disease of accessible NF mediation comprises the unify cancer of skin (like melanoma and Kaposi sarcoma) of mammary gland, thyroid, colon, lung, ovary, skin, muscle, pancreas, prostate, kidney, genitals, blood, immune system (for example spleen, thymus and bone marrow), brain, peripheral nervous system according to the present invention.

Answering of neuronal function in infringement or unusual tissue or relevant with infringement or unusual tissue Former or normalization

The recovery or the normalization of neuronal function in infringement or unusual tissue or relevant with infringement or unusual tissue are provided among the present invention in one aspect.Said tissue can be brain tissue or the outer tissue of brain, for example skin, bone, eye or muscular tissue.

Of the present invention this on the one hand can, for example, unite use with the recovery of nerve after operation, incised wound, wound, unexpected injury, contusion, scratch, burn, cold injury, the fracture.

Wound healing

Auxiliary wound healing is provided among the present invention in one aspect.Wound can be any skin lesion, comprises chronic (for example ulcer property) skin lesion and polarity skin lesion.These sick reasons of decreasing are a lot of and variable.In general all skin lesions can use the useful processing of the present invention.

Measurement comprises the closing speed of wound with the aspect of the wound healing of the quality of assessment healing; The reproduction speed of skin histology on the wound; The color of the wound of the healing relevant with cutaneous pigmentation on every side; The mechanical strength of the wound of the healing relevant with surrounding skin intensity; Cicatricial tissue or other skin histologies with unusual texture or rugosity are being retained in the degree on the wound after the healing to greatest extent; Wound stops to ooze out or the exudate flow reduces the used time; The physical appearance of wound or effluent and smell, and in the agglutination different time pain, scratch where it itches or other uncomfortable degree.

Contrast all these standards, the present invention provides with prior art and handles the benefit of comparing.Do not have must adding of external source NF, the stable state of self regulating of curee's natural NF is expected at the human and non-human mammal skin lesion of useful influence under all employed standards.

According to the of the present invention dose of processing that can be used for wound by external or systemic application.If external is used, they can be sent from any suitable compositions or structure, for example are used for the dressing of wound or are applied to emulsifiable paste or other preparations of wound.The other details of delivery system is provided in hereinafter.

The Ankang and the general health of promotion or aid in tissue

The present invention provides in yet another aspect and promotes muscle and its hetero-organization to restore and raising endurance and muscle endurance (for example competitiveness or noncompetitive motion) and the tired sensation of minimizing from taking exercise, work or consuming.

And usually, the Ankang of tissue and general health (in brain with brain outside both) can be assisted according to the present invention.

In an example, will improve the healthy and safe and comfortable sensation of replenishing of new Skin Cell and general so aid in skin or eye according to of the present invention dose of cosmetics, eye or dermatological applications to skin.Referring to for example Alber, people such as K.M., Neuroscientist 2007,13, the 371-382 page or leaf.The method according to this invention relates to the stable state of self regulating of skin NF, and it avoids health is used toxic agents, and the natural NF that the substitute is adjusting curee oneself is so that handle.

Although be not exclusively, these purposes are non-therapeutic normally, and main targeting is healthy human.

Mammal

Useful except the mankind are handled, the present invention is useful equally in many mammals that can receive neurological and psychology/psychiatry patient's condition influence equally.These mammals comprise inhuman primates (for example apes, monkey class and Prosimiae) for example at the zoo in; Companion animals is cat class or dog class for example; Work or motion animal be dog class, horse class and pony class for example; Farm-animals is Swine, sheep, goat class, deer, bovine and domestic animal for example, and laboratory animal for example Lagomorpha or Rodents (such as rat, mice, hamster, gerbil jird or Cavia porcellus).

When the disease that remains to be processed, the patient's condition, speciality or function when being unique, should be understood that so the mammal that remains to be processed is human to the mankind.Identical understanding is applied to any other mammalian species respectively, if the disease that remains to be processed, the patient's condition, feature or function are unique to those species.

Agent

The activating agent that this paper uses usually but be not the molecular weight that has basically less than about 800, for example less than about 700, for example less than about 600, for example less than about 500, for example less than about 450.

According to the standard name from steroid class chemistry, 6 yuan of rings of left-hand side are named as the A ring, with the ring called after B ring of A ring adjacency.According to the standard name from steroid class chemistry, carbon atom is numbered shown in hereinafter, so that the line that condenses between the ring is present between 5 and 10 carbon atoms equally.

In A/B cis steroidal furostan/furan steroid alkene or spirostane/spirostene ruscogenin, become the β location 5 with the substituent group or the hydrogen atom of 10 carbon atoms, it is positioned at the plane (top) of molecule.

This plane kinking that has molecule seems as the hereinafter effect of the pharmacophoric group in three-dimensional picture with generation.Substituent group or hydrogen atom on 10 carbon atoms are marked as " a " in picture, and substituent group or hydrogen atom on 5 carbon atoms are marked as " b "; C ring only part is showed:

This is an A/B cis primitive.

The instance that is disclosed in A/B cis furostan/furan steroid alkene and spirostane/spirostene ruscogenin and their derivative form among WO-A-99/48482, WO-A-99/48507, WO-A-01/23407, WO-A-01/23408, WO-A-02/079221, WO-A-03/082893, WO-A-2005/105825 and the WO-A-2006/048665 can be used as the activating agent that is used for purposes of the present invention and is mentioned especially.Chemical compound and the independent chemical compound of certain set that is disclosed in said type chemical compound of representative A/B cis furostan/furan steroid alkene or spirostane/spirostene ruscogenin and its ester, ether, ketone and glycosylated form in these publications incorporated this paper by reference into.

The ester of A/B cis furostan/furan steroid alkene and spirostane/spirostene ruscogenin, ether, ketone and glycosylated form so consequently one or more esters, ether, ketone and glycosylation group can be present in the molecule.In general, ester, ether, ketone or glycosylation group can use traditional chemical synthesis process partly to form at any one or a plurality of OH of A/B cis spirostane/spirostene ruscogenin.

According to the instance of activating agent of the present invention is the A/B cis-compound by following formula representative: the formula I among the WO-A-01/23406 (disclosed PCT application the 6th page); Formula II among the WO-A-01/23406 (disclosed PCT application the 7th page); Formula I among the WO-A-01/23407 (disclosed PCT application the 6th page); Formula II among the WO-A-01/23407 (disclosed PCT application the 6th page); Formula I among the WO-A-01/23408 (disclosed PCT application the 6th page); Formula I among the WO-A-01/49703 (disclosed PCT application the 7th page); Formula II among the WO-A-02/079221 (disclosed PCT application the 6th page); Formula I among the WO-A-03/082893 (referring to the page 4 of disclosed PCT application); Formula II among the WO-A-03/082893 (referring to the page 4 of disclosed PCT application); Formula III among the WO-A-03/082893 (referring to the page 5 of disclosed PCT application); Formula II among formula I among the EP-A-1024146 (referring to the page 4 of disclosed EP application) and the EP-A-102416 (apply for referring to disclosed EP the 8th page).

For example, molecule chinaroot greenbrier sapogenin and smilagenin are useful for purposes of the invention activating agents with their corresponding esters, ether, ketone and Saponin (glycosylation) derivant.The chemical compound zhimusaponin BII that is A/B cis furostan Saponin is useful for purposes of the invention activating agent.

Be used for other useful activating agents of the present invention and comprise table chinaroot greenbrier sapogenin, table smilagenin, metagenin, samogenin, the cautious ruscogenin of buchu (diotigenin), strange land cautious ruscogenin difficult to understand, texogenin (texogenin), yonogenin, mexogenin and markogenin and their corresponding esters, ether, ketone and saponin derivative.

Crystal that said activating agent is can be any suitable or noncrystalline form and any suitable anhydrous, hydration or solvation form are used.The further details of these forms of chinaroot greenbrier sapogenin and smilagenin and their derivant provides in WO-A-2005/105825 that particular reference considers is pointed to and WO-A-2006/048665.

Lipid can comprise especially 3 esters for example carboxylate (like cathylate (ethoxy carbonyl oxygen base); Acetas; Succinate; Cinnamate; Ferulic acid ester; Propionic ester; Butyrate; Isobutyrate; Valerate; Isovalerate; Alkyl caproate; Dissident's acid esters; The diethacetic acid ester; Caprylate; Decanoin; Laurate; Myristinate; Cetylate; Stearate; Benzoate; Phenylacetate; Phenpropionate; Cinnamate; The Nitrodracylic acid ester; 3; 5-dinitrobenzoic acid ester; To chlorobenzene carboxyphenyl ester; 2; 4-chlorobenzene carboxyphenyl ester; To bromobenzene carboxyphenyl ester; Between bromobenzene carboxyphenyl ester; To the methoxybenzene carboxyphenyl; The phthalyl ester; Glycinate; Alanine ester; L-valine ester; Phenylalanine ester; The isoleucine ester; The methionine ester; Arginine ester; The agedoite ester; Aspartate; Cysteine ester; Glutamate; The histidine ester; The lysine ester; Proline ester; Serine ester; The threonine ester; The tryptophan ester; Tyrosine ester; Fumarate; Maleate), phosphonate ester and sulphonic acid ester.

Ether can be particularly including 3 ethers alkoxyl derivatives (for example methoxyl group, ethyoxyl, positive propoxy, secondary propoxyl group, n-butoxy, sec-butoxy, tert-butoxy) for example.

Ketone (Saponin ketone (sapogenone)) is the 3-ketone derivatives of corresponding ruscogenin normally, although other ketone derivatives that form at the different OH load carbon atoms place of loop systems also are possible.The instance of 3-ketone Saponin ketone comprises pariglin ketone (sarsasapogenone), smilagenone, table pariglin ketone (episarsasapogenone) and table smilagenone.

The instance of the saponin that is fit to comprises that wherein the carbon atom of 3-position (is R ₃The carbon that connects) be loaded with replacement R ₃The O-sugar moieties, for example, monosaccharide, disaccharide or trisaccharide or higher polysaccharide or its acidylate form.These sugared examples of groups comprise and are selected from glucose; Mannose; Fructose; Galactose; Maltose; Cellobiose; Sucrose; Rhamnose; Xylose; Arabinose; Fucose; Quinovose; Celery sugar; Lactose; Galactose-glucose; Glucose-arabinose; Fucose-glucose; Rhamnose-glucose; Glucose-glucose-glucose; Glucose-rhamnose; Mannose-glucose; Glucose-(rhamnose)-glucose; Glucose-(rhamnose)-rhamnose; Glucose-(glucose)-glucose; The glycosyl group of galactose-(rhamnose)-galactose and its acidylate (for example acetylation) derivant.

Pseudo-saponin (unit) class is the derivant of the ring opening of corresponding spirostane/spirostene ruscogenin or Saponin, and wherein the F ring is opening and locking.Pseudo-saponin (unit) class can have the saturated or unsaturated of C20-C22 key place.Saturated form is called as " the pseudo-saponin of dihydroxy (unit) class " form sometimes.

Being used for activating agent of the present invention can be used separately or with desired combination.

Other adjuvant or auxilliary composition

The compositions of using among the present invention can comprise one or more adjuvant and/or one or more auxilliary compositions if desired, as relevant with compositions and application route hereinafter in greater detail.

Especially, can use chemical compound, the energy that can change into the TCA intermediate in metabolism adjuvant, the chemical compound (ketogenic compounds) that increases the ketoboidies level, tricarboxylic acids (TCA) intercycle body, the body to strengthen chemical compound or its any mixture.

The metabolism adjuvant comprises vitamin (for example vitamin E); Mineral, the chemical compound that antioxidant is relevant with other (for example ascorbic acid, biotin, calitriol, vitamin B12, folic acid, nicotinic acid, pantothenic acid, vitamin B6, vitamin A, axerophthal (retinal), tretinoin, riboflavin, thiamine, alpha-tocopherol, vitamin K1, Menaquinone K6, calcium, magnesium, sodium, aluminum, zinc, potassium, chromium, vanadium, selenium, phosphorus, manganese, ferrum, fluorine, copper, cobalt, molybdenum, iodine or its any combination.

Ketogenic compounds increases receptor's endogenous lipid metabolism (oxidation) usually and increases blood ketone level thus, and comprises for example C _3-8Ketone is such as acetone, D-beta-hydroxybutyric acid, the metabolic precursor thereof of D-beta-hydroxybutyric acid (for example acetoacetyl based precursor such as acetoacetyl-1,3 butylene glycol, acetoacetyl-D-beta-hydroxybutyric acid and acetoacetyl base glycerol; Ester is D-beta-hydroxybutyric acid and the pure ester of monohydroxy, dihydroxy or trihydroxy for example; Or the polyester of D-beta-hydroxybutyric acid for example has about 2 and gathers the D-beta-hydroxybutyric acid such as about 3 to about 10 the multiple D-of gathering beta-hydroxybutyric acids or terminal oxidation to about 100 repetitions), the metabolic precursor thereof of acetoacetic acid or its any combination.

The TCA intermediate comprises citric acid, equisetic acid, 1-Hydroxy-1,2,3-propanetricarboxylic acid., α-Tong Wuersuan, succinic acid, fumaric acid, malic acid, oxo acetic acid or its any combination.

The chemical compound that can change into the TCA intermediate in the body comprises 2-ketone-hydroxyl propanol, 2; 4-dihydroxy butanols, 2-ketone-4-hydroxyl butanols, 2,4 hydroxybutyric acid, 2-ketone-4 hydroxybutyric acid, aspartate, list and two-alkyl-methyl-oxalacetic ester, pyruvate, glucose-6 phosphate ester or its any combination.

Energy strengthens chemical compound and comprises for example coenzyme CoQ-10, creatine, creatine derivant, L-carnitine, n-acetyl-carnitine, L-carnitine derivant or its any combination.These chemical compounds produce through many mode enhanced energy.Carnitine will increase the metabolism of fatty acid.Play the effect of electron carrier in the electron transfer process of CoQ-10 in mitochondrion.Correspondingly, add these chemical compounds and activating agent for example medium chain triglyceride (MCT) will increase metabolism and render a service, especially in the individuality that can be removed nutrition.

Said adjuvant can metabolic precursor thereof when existing form for example provide in therapy or nutrition, to use with one or more cationic complex or as salt.Cation comprises sodium, potassium, magnesium, calcium salt with the instance of typical physiology salt, and for example L-lysine, L-arginine, methyl glucoside amine or other materials as known in the art come balance to physiological equilibrium's ion of cation through forming salt complex in each instance.The preparation of these metabolic precursor thereof and use are described among WO-A-98/41201 and the WO-A-00/15216, and its open method is incorporated this paper by reference into.

Compositions and application route

Activating agent can comprise the form of the compositions of activating agent and any suitable other composition and used.Compositions can for example be pharmaceutical composition (medicine), food, food supplement or beverage.This compositions can comprise the mixture of specific chemical compound and/or the acceptable ester of their physiology, amide, salt, solvate, analog or other derivants that are fit to.Usually, mention among this paper a kind of activating agent and/or compositions other compositions exist in two or more the existence of mixture that comprises these agent and/or composition in its scope.

Pharmaceutical composition can through any suitable route use, include but not limited to oral, the nose stomach, rectum, transdermal, parenteral (in for example subcutaneous, intramuscular, intravenous, the marrow and the injection or the infusion of Intradermal), intranasal, saturating mucosa, inculcate, vagina, external, through cheek with the Sublingual.

Many as in the activating agent, using the site can be the typical characteristic of the use of some micromolecule lipophilic agent away from the mammiferous brain that remains to be processed, said dose is passed blood brain and/or blood-nerve barrier through the blood migration.

Term " pharmaceutical composition " means the compositions that the character that depends on method of application and dosage form comprises activating agent and comprises additionally pharmaceutically acceptable carrier, diluent, adjuvant, adjuvant or vehicle such as preservative agent, filler, disintegrating agent, buffer agent, preservative agent, penetration enhancers, humidizer, emulsifying agent, suspending agent, sweeting agent, flavoring agent, aromatic, antibacterial agent, antifungal, lubricant and dispersant in the context of the present invention.The dosage form that is fit to comprises for example tablet, lozenge, powder, elixir, syrup, and liquid preparation comprises suspension, spray, inhalant, tablet, dragee, Emulsion, solution, granule, capsule and suppository and be used for injection liquid preparations and comprise Liposomal formulation.Technology and preparation usually can be at Remington, Pharmaceutical Sciences (pharmacy), and Mack Publishing Co., Easton, PA finds in the latest edition.

Term used herein " food ", " food supplement ", " beverage " and " beverage fill-in " have the normal implication of these terms and are not restricted to pharmaceutical preparation.These compositionss are applicable to oral absorption.Fill-in compositions (for example food supplement or beverage fill-in) is arranged to add to F&B and is ingested with them.Food can comprise usually gives birth to hot material for example fat, oils and fats and carbohydrate and protein and mineral and fibre source.The instance of compositions comprises the food that is the basis with milk product, frumentum, vegetable, meat, fish, poultry or fruit.The instance of beverage comprises carbonated beverage and noncarbonated beverage products, and fruit juice, infusion beverage is coffee or tea such as herbal tea, fruit juice, SEN CHA or India or Chinese tea for example.Compositions can comprise the composition in milk or milk source, for example milk powder and/or lactose and/or casein.The composition in milk or milk source preferably derives from milch cow or goat.The milk that can use plant origin is bean milk for example.Edible compositions can comprise the component of one or more fermentations.Compositions can comprise Yoghurt.Food supplement can for example contain vitamin, mineral, caffeine, ephedra alkaloid.

Orally administered composition

The instance of the form of taking in that is fit to includes but not limited to solid, has the dosage form of liquid, powder or solid core; Can chew or oral disintegrating tablet; The plaster applying a plaster for therapeutic purpose sheet; The natural gum tablet; The coated granules that has the salivation derivant in foam tablet and coating and/or the granulating substrate.In one embodiment, dosage form is that design is to comprise a certain composition for example hereinafter solid, semisolid or the fluid composition of the specific scheduled volume of defined active component (being dosage).The dosage form that is fit to can be comprise be used for Orally administered, use or those pharmacy drug delivery system of mucosal delivery through cheek; Or the compositions that is used to send mineral, vitamin and other dietetic products, oral cavity nursing agent, flavoring agent and analog.In one embodiment, dosage form of the present invention is counted as solid; Yet they can comprise liquid or semi-solid components.In another embodiment, dosage form is the Orally administered system that is used for to the mankind's gastrointestinal tract delivery of pharmaceutically active composition.The adjuvant that is fit in the compositions can comprise analgesic; Antiinflammatory; Anti-arthritic; Anesthetis; Hydryllin; Cough medicine; Antibiotic; Anticarcinogen; Anti-allergic agent; Anti-infective; Antiviral agent; Anticoagulant; Antidepressants; Antidiabetic; Bendectin; Antiflatulent; Antifungal agent; Spasmolytic; Appetite suppressant; Bronchodilator; Cardiovascular agents; Central nervous system agent; Central nervous system stimulant; Immune system analeptic; Decongestant; Diuretic; Expectorant; The gastrointestinal tract agent; The migraine preparation; Carsick product; Mucolytic; Muscle relaxant; The osteoporosis preparation; Polydimethylsiloxane; Breathe agent; Sleeping pill; Urinary tract agent and composition thereof.The oral cavity nursing agent that is fit to can exist, for example breath freshener, dental bleaching agent, antimicrobial, tooth mineralizer, decayed tooth inhibitor, local anesthetic, mucosa protective agent (mucoprotectant) and analog.The flavoring agent that is fit to comprises menthol, Oleum menthae, mint flavouring, fruit flavor, chocolate, Rhizoma et radix valerianae, bubble gum flavor, coffee flavour, sweet wine spice and combination and analog.The instance of the gastrointestinal tract agent that is fit to that also can exist comprises antacid for example calcium carbonate, magnesium hydroxide, magnesium oxide, magnesium carbonate, aluminium hydroxide, sodium bicarbonate, mincid; Irritant purgative for example bisacodyl, cascara sagrada, dantron, Folium Sennae, phenolphthalein, Aloe, Oleum Ricini, castor oil acid and dehydrocholic acid with and composition thereof; H ₂Receptor antagonist is famotidine, ranitidine, cimetidine, nizatidine for example; Proton pump inhibitor is omeprazole or lansoprazole for example; The gastrointestinal cytoprotection agent is sucralfate and misoprostol for example; The gastrointestinal tract activator is prucalopride for example; The antibiotic that is used for helicobacter pylori (H.pylori) is clarithromycin, amoxicillin, tetracycline and metronidazole for example; Diarrhea is diphenoxylate and loperamide for example; Glycopyrrolate; Bendectin is ondansetron for example; Analgesic is mesalazine for example.Be selected from analgesic, the agent of anti-inflammatory agent and febrifuge also can exist: for example NSAID (non-steroidal anti-inflammatory drug) (NSAID) comprises that propanoic derivatives for example: ibuprofen, naproxen, ketoprofen and analog; Acetogenin is indomethacin, diclofenac, sulindac, Tolmetin and analog for example; Fenamic acid derivative is for example: mefenamic acid, meclofenamic acid, flufenamic acid and analog; The xenyl carboxylic acid derivates is for example: diflunisal, flufenisal and analog and former times the health class for example: piroxicam, sudoxicam, isoxicam, meloxicam and analog.In one embodiment, auxilliary active (coactive) composition can be selected from propanoic derivatives NSAID: for example ibuprofen, naproxen, flurbiprofen, fenbufen, fenoprofen, indoprofen, ketoprofen, fluprofen, pirprofen, carprofen, oxaprozin, pranoprofen, suprofen and its pharmaceutically acceptable salt, derivant and combination.In another embodiment of the present invention, active component can be selected from acetaminophen, aspirin, ibuprofen, naproxen, ketoprofen, flurbiprofen, diclofenac, cyclobenzaprine, meloxicam, rofecoxib, celecoxib and its pharmaceutically acceptable salt, ester, isomer and mixture.

In another embodiment, auxilliary activating agent can be selected from pseudoephedrine, phenylephrine, phenylpropanolamine, chlorphenamine, dextromethorphan, diphenhydramine, guaifenesin, astemizole, terfenadine, fexofenadine, loratadine, Desloratadine, doxylamine, norastemizole, cetirizine, benzocaine, its mixture and its pharmaceutically acceptable salt, ester, isomer and mixture.In another embodiment, auxilliary active component can be other activating agents of methylphenidate, modafinil and suitable hyperkinetic syndrome or attention deficit hyperactivity disorder; Oxibutynin; 'Xiduofeng ' and cyclobenzaprine.Active component is present in dosage form of the present invention to treat effective amount, and said treatment effectively amount is the amount that the desired treatment of Orally administered generation is responded and can be confirmed by those of skill in the art in this area at an easy rate.When confirming this tittle, as as known in the art, the specific active ingredient of being used, the bioavailability characteristic of active component, dosage regimen, patient's age and body weight and other factors must be considered.In one embodiment, dosage form comprises the active component at least about 85 percentage by weights.Active component can be present in the dosage form in any form.For example, active component can be on molecular level be disperseed for example fusion or is dissolved in the dosage form, or can particulate form, and it is coating or not coating then.If active component is particulate form, granule (no matter coating or not coating) have usually about 1 micron to about 2000 microns average particle size.In one embodiment, these granules are to have about 1 micron crystal to about 300 microns average particle size.In also having another embodiment, these granules are to have about 50 microns to about 2000 microns, for example from about 50 microns to about 1000 microns or pellet or granule from 100 microns to about 800 microns average particle size.

In one embodiment, Orally administered composition of the present invention is a food compositions, for example the mankind or pet food.In certain embodiments, compositions is a food compositions, and except activating agent, also comprising every kind is about 15-50% albumen, about 5-40% fat, about 15-60% carbohydrate, the 5-10% ash on basis with the dry weight, and has the water capacity of 5-20%.In certain embodiments, the food expection provides necessary completely diet needs.Also provide as snacks, house pet snack (for example cookies), nutrition bar and be used for food product or the useful compositions of other forms of dietary supplements, comprise tablet, capsule, gel, paste, Emulsion, lozenge and analog as hereinafter being discussed.Randomly, food compositions can be anhydrous composition (coarse grain that for example is used for pet food), half wet compositions, moisturizing compositions or its any mixture.

Compositions of the present invention can be the food product that is in particular the human consumption preparation.These will comprise the expection supply mankind's the necessary diet needs and the food and the nutriment of other human dietary supplements.In one embodiment, for the food product of human consumption preparation is completely and nutritive equilibrium, and in other, the dietary supplements of their expection conducts and the balance relevant use of diet good or preparation.

Compositions can be a food supplement, for example gravy, drinking water, beverage, concentrated solution, gel, yoghourt, powder, granule, paster, suspension, chaw, delicacies (morsel), snack, snacks, pill, pill, capsule, tablet or any other delivery form.Term " food supplement " comprises dietary supplements.Dietary supplements can be specific species or even for example companion animals or human consumption and special preparation of individual animals.In one embodiment, dietary supplements can comprise relatively and to concentrate dose of active so that fill-in and can be applied to animal or can dilution before being applied to animal in a small amount.In certain embodiments, dietary supplements or other comprise active compositions and can mix with water or analog before the animal being applied to, for example with adjustment dosage, so that its better to eat or permission is with more frequent the using of littler dosage.

Compositions of the present invention can be cooled or be freezing.Activating agent can with other composition premixs of compositions so that needed favourable amount to be provided; Can be by emulsifying; On coating to pet food composition, dietary supplements or the food product, or can add to compositions before consuming it or it is offered animal (for example using powder or mixture) for the human consumption preparation.

In one embodiment, compositions comprises activating agent with the amount that animal or human's apoplexy due to endogenous wind of effectively being used in compositions has desired physiology or psychology or ethological effect.For pet food be the food product of human consumption preparation, as the amount of the activating agent of the percentage ratio of compositions compositions about 1% to 30% who with the dry is the basis, although littler or higher percentage ratio can be provided.In different embodiments; Amount be with the dry be the basis compositions about 1.0%; 1.5%; 2.0%; 2.5%; 3.0%; 3.5%; 4.0%; 4.5%; 5.0%; 5.5%; 6%; 6.5%; 7%; 7.5%; 8%; 8.5%; 9%; 9.5%; 10%; 10.5%; 11%; 11.5%; 12%; 12.5%, 13%; 13.5%; 14%; 14.5%; 15%; 15.5%; 16%; 16.5%; 17%; 17.5%; 18%; 18.5%; 19%; 19.5%; 20%; 20.5%; 21%; 21.5%; 22%; 22.5%; 23%; 23.5%; 24%; 24.5%; 25%; 25.5%; 26%; 26.5%; 27%; 27.5%; 28%; 28.5%; 29%; 29.5%; 30% or more.Can be with the dietary supplements preparation to comprise the activating agent of several times of high concentrations; Being applied to animal or human's class or dilution before using with tablet, capsule, concentrated solution or other similar dosage forms; For example through in water, diluting, spray or be sprayed at house pet or the human food goes up and the using of other similar manners.For dietary supplements, activating agent can directly be applied to animal or human's class separately or be directly applied to the conventional food of animal or human's class.

Compositions can randomly comprise supplementation material for example mineral, vitamin, salt, condiment, pigment and antiseptic.The limiting examples of the mineral that replenishes comprises calcium, phosphorus, potassium, sodium, ferrum, chloride, boron, copper, zinc, magnesium, manganese, iodine, selenium and analog.The limiting examples of the vitamin that replenishes comprise in vitamin A, the category-B vitamin any, vitamin C, vitamin D, vitamin E and vitamin K, comprise aforesaid various salt, ester or other derivants.Also can comprise other dietary supplements, any form of nicotinic acid, pantothenic acid, inulin, folic acid, biotin, aminoacid and analog for example, with and salt and derivant.In addition, compositions can comprise favourable long-chain polyunsaturated fatty acid for example (n-3) and/or (n-6) fatty acid, arachidonic acid, eicosapentaenoic acid, clupanodonic acid and docosahexenoic acid with and the combination.

The compositions that this paper provides randomly comprises and promotes or to keep general neurological healthy or further strengthen one or more supplementation material of cognitive function.These materials comprise for example choline, Phosphatidylserine, alpha-lipoic acid, CoQ10, acetyl-L-camitine and for example comprise a herb ingredients or the extract from one or more compositions of the plant of dripping flower (Leucojum aestivum) such as Semen Ginkgo (Ginko biloba), Herba Bacopae monnieri (Bacopa monniera), the Scolopendra of crawling (Convolvulus pluricaulis) and/or snow.

In different embodiments, it is from 15% to about 50% the crude protein on basis that food that this paper provides or food/meal supplement compositions preferably comprises with the dry.The crude protein material comprises one or more albumen in animal, plant or other any source from no matter.For example, for example Semen Glycines, Semen Gossypii and Semen arachidis hypogaeae are the purposes that is suitable for this paper to vegetable proteins.Animal and milk protein be casein, albumin and meat proteins for example, comprises that Carnis Sus domestica, lamb meat, Equus caballus (L.), poultry meat, the flesh of fish or its mixture are useful.

It is about 5% to about 40% the fat on basis that compositions can further comprise with the dry.Compositions also can further comprise carbohydrate source.It is about 15% to about 60% the carbohydrate on basis that compositions comprises with the dry usually.These examples of carbohydrates comprise corn or grain for example Oryza sativa L., corn, Sorghum vulgare Pers., Herba Medicaginis, Fructus Hordei Vulgaris, Semen sojae atricolor, Brassica campestris L, Herba bromi japonici, Semen Tritici aestivi or its mixture.Compositions also randomly comprises other compositions, and it comprises the for example anhydrous milk surum of carbohydrate and other breast system product or side-product.

Compositions also can comprise at least a fibre source.In the various soluble or insoluble fiber that is suitable in food or feedstuff, using any can be utilized and these are that those skilled in the art are known.The fibre source that is fit to comprises oligofructose, mannan oligofructose, Semen Glycines fiber, arabinogalactan, galacto-oligosaccharide, araboxylan or its mixture outside beet pulp (from Radix Betae), Radix Acaciae senegalis, talha, Psyllium, Testa oryzae, carob, citrus pulp, pectin, the short chain oligofructose.Selectively, fibre source can be fermentable fiber.Be described the immune benefit that provides companion animals before fermentable fiber.Provide prebiotics (prebiotic) compositions fermentable fiber or other compositionss known to those skilled in the art can be impregnated in compositions equally to help to strengthen the benefit that provides to the immune system of animal by the present invention to strengthen prebiotics microbial growth in the intestinal.In addition, prebiotics microorganism such as lactobacillus (Lactobacillus) or bacillus bifidus (Bifidobacterium) species for example, can be added to compositions.

In another embodiment, Orally administered composition of the present invention is a carbonated beverage compositions, comprises concentrate thus.These compositionss can prepare through the method for knowing in this area.

In this embodiment, because carbon dioxide (in water, forming carbonic acid), beverage is normally tart.Yet possible for this beverage is " by acidify ", promptly is conditioned so that it comprises the other acid of the said type that remains in " intensive " beverage, to be found.Instance can comprise that phosphoric acid and acid condiment (being called " wholesome acid " sometimes) are such as citric acid, maleic acid, fumaric acid and tartaric acid.Fruit, fruit juice and fruit extract comprise acid condiment, and the beverage that therefore contains these compositions can be considered to acidifying.

Beverage can be nonalcoholic.Instance comprises cola drink, Citrus beverage, Fructus Citri Limoniae beverage, lemonade, nourishing water, root medicated beer, Rhizoma Zingiberis Recens malt liquor and ginger beer.

Beverage can be alcoholic, has 3-9%wt/wt ethanol usually.Instance comprises applejack and what is called " Eaux-De-Vie ", and it has fruity, normally the carbonated admixture of vodka or other alcoholic beverage.Beverage can be alcoholic a little, has 0.1-3%wt/wt ethanol usually.Instance comprises root medicated beer, ginger beer and the lemonade that mixes wine (shandy) and some fermented type.

Soda pop can be the non-dairy product, does not for example have the beverage of suckling or not having yoghourt.Soda pop can be fat free basically.

Beverage can be the beverage that the water with seasoning is the basis.

Soda pop can be clarifying or muddy or chaos or opaque.

Soda pop can contain vitamin, for example one or more in A, B, C, D, E and the K group vitamin.Can add the vitamin the vitamin that for example exists in the fruit juice except other components.Vitamin B that can be water-soluble and C are fit closely beverage ingredient.Lipid-soluble vitamin A, D, E and K are poor slightly.Preferably the vitamin E or derivatives thereof is not present in the beverage.Preferably vitamin A and K or derivatives thereof do not exist in beverage.

Soda pop can contain sweeting agent.Sweeting agent can be natural or synthetic sweeting agent, for example sugar, corn syrup, sugar alcohol (for example Sorbitol, xylitol, mannitol, maltose alcohol or hydroxyl isomaltulose) or intensive sweetener (for example glucide, sucralose, neotame, acesulfame potassium or aspartame) or its any combination.

Topical composition

In another embodiment, compositions of the present invention is a topical composition, for example the compositions of cosmetics, eye or dermatological.

Preparation in any suitable manner is used for the topical composition of sending of activating agent.Topical composition can be formulated as wound adjuvant or other medical applications systems in a conventional manner.

Compound active agent described herein can be on one or more cosmetics and/or dermatological acceptable carrier and therefore randomly the other treatment composition prepare and send.Carrier should be can be received, because they are compatible with any other composition of compositions and are harmless to its receptor.Carrier also can reduce by any side effect of not expecting of said dose.These carriers or vehicle composition are as known in the art.Referring to Handbook of Cosmetic Science and Technology (cosmetic science and technical manual) Taylor & Francis Group, 2006, it incorporates this paper by reference in full into.

Be used for that compositions that external according to the present invention uses can be used for the part and/or system uses, the active component and zone of using and the frequency that depend on wherein to be provided.Therefore, should be regarded as to the following discussion of external preparation with degree descriptive system preparation comprising activating agent that can the external systemic application.

The compositions that is used for using in the external that combination of the present invention is used can be impregnated in usually any medicine, cosmetics, eye or dermatology preparation used and that can exist in a variety of forms.For example, being used for the compositions that external uses can be solution, Water-In-Oil (W/O) type emulsion, oil-in-water (O/W) type emulsion or multiple emulsion for example W/O/W (W/O/W) or Water-In-Oil bag oil (O/W/O) emulsion, aqueous dispersions or fat dispersion liquid, gel, emulsifiable paste, solid bar or aerosol.According to Emulsion of the present invention, for example,, be easily and comprise for example fat, oil, wax and/or other lipids with the form of emulsifiable paste, lotion or the breast of making up, and water and be normally used for one or more emulsifying agents of the preparation of this type like them.

In certain embodiments, be used for the composition that compositions that external according to the present invention uses can be depending on them and be used as for example protective skin cream, facial milk cleanser, sun-proof lotion, nourishing cream, day cream or late frost and analog.

Be used for the compositions that external uses and comprise effective ingredient, cosmetics supplement and/or cosmetics additive on the cosmetics that are generally used for these preparations.These comprise for example antioxidant; Antiseptic; Antibacterial; Thickening agent; Filler; Defoamer; Spice; Quintessence oil; Pigment (fumed silica for example; Microfoam pigment such as oxide and silicate comprise the ferrum oxide of choosing coating wantonly; Titanium dioxide; Boron nitride and barium sulfate); Ceramide (no matter being the function plan thing of natural material or natural neural acyl); Surfactant; Emulsifying agent; Phospholipid; Cholesterol; Phytosphingosine; Other active component for example vitamin or protein (such as retinyl palmitate or retinyl acetate; Vitamin B for pantothenylol and derivant thereof; Vitamin E for tocopherol acetas; Be the vitaminF of esters of polyunsaturated fatty acids) such as the gamma-Linolenic acid ester; Opacifier (comprising chemical sunscreen agents and dispersive physical sunscreen); Stabilizing agent; Anthelmintic; Alcohols; Plasticizer; Polyhydric alcohol; Polymer; Foam stabiliser; Electrolyte; Organic solvent; Silicone derivative; Humidizer and/or wetting agent; Oils and fats; Oil; Wax; Water; Salt; Proteolysis or cutin hydrolysing activity material and analog.These additives can be present in and be used for Dermatology or the cosmetic composition that external is used.

Like what above write down, except being used for activating agent that external sends, topical composition of the present invention also can comprise one or more other activating agents or the material that beneficial effect is provided.For example, in specific embodiment, topical composition can comprise sunscreen product.These preferably comprise at least a other UVA filtrate and/or at least a UVB filtrate and/or at least a inorganic pigment except active component used according to the invention.

The UVB filtrate can be soluble in oil or water.The instance of soluble material for example is in oil: 3-benzylidene Camphora and derivant thereof; 3-(4-methylbenzene methylene) Camphora for example; 4-aminobenzoic acid derivative, preferably 2-ethylhexyl 4-dimethylaminobenzoic acid ester, amyl group 4-dimethylaminobenzoic acid ester; Cinnamate, preferably 2-ethylhexyl 4-Methoxycinnamate, isopentyl 4-Methoxycinnamate; Salicylate, preferably 2-Ethylhexyl salicylate, 4-isopropyl benzyl salicylate, homosaligenin (homomethyl salicylate); Benzophenone derivates, preferably 2-hydroxyl-4-methoxy benzophenone, 2-hydroxyl-4-methoxyl group-4 '-menthyl benzophenone, 2,2 '-dihydroxy-4-methoxy benzophenone; Benzylidene malonate, preferably two (2-ethylhexyl) 4-methoxybenzene methylene malonate; 2,4,6-triphen amido-(to carbonyl-2 '-ethyl-1 '-hexyloxy)-1,3,5-trizane.

Soluble favourable material is in water: 2-Phenylbenzimidazole-5-sulfonic acid and its salt, for example sodium, potassium or triethanolamine salt, the sulfonic acid of benzophenone, preferably 2-hydroxyl-4-methoxy benzophenone-5-sulfonic acid and its salt; The sulfonic acid of 3-benzylidene Camphora is for example such as 4-(2-oxo-3-bornyl subunit-methyl) benzenesulfonic acid, 2-methyl-5-(2-oxo-3-bornyl ylidenylmethyl) sulfonic acid and their salt.Natively, above mention can be used according to the invention the tabulation of UVB filtrate be not mean restrictive.

The instance of UVA filtrate that can be used according to the invention comprises the DBM derivant, 1-(4 '-tert-butyl-phenyl)-3-(4 '-methoxyphenyl) propane-1 especially, 3-diketone and 1-phenyl-3-(4 '-isopropyl phenyl) propane-1, the 3-diketone.

The instance of inorganic pigment that can be used according to the invention comprise oxide and their mixture of titanium, zinc, ferrum, zirconium, silicon, manganese, aluminum, cerium and wherein oxide be the modification of activating agent.Especially, preferably, they are to be the pigment on basis with titanium dioxide.

Favourable antioxidant that can be used according to the invention is that concerning cosmetics and/or eye and/or dermatological applications, be fit to or conventional all that antioxidant.Antioxidant advantageously is selected from the group of being made up of following: aminoacid (glycine for example; Histidine; Tyrosine; Tryptophan) and their derivant; Imidazoles (for example urinating Canis familiaris L. acid) and their derivant; Peptide is D for example; L-carnosine (D; The L0 carnosine); The D-carnosine; L-carnosine and their derivant (for example anserine); Carotenoid; Carotenoid (alpha-carotene for example; Beta-carotene; Lycopene) and their derivant; Aurothioglucose; Propylthiouracil and other mercaptan (thioredoxin for example; Glutathion; Cysteine; Cystine; Cystamine and their glycosyl; The N-acetyl group; Methyl; Ethyl; Propyl group; Amyl group; Butyl and lauryl; Palmityl; Oil base; γ-Ya oil base; Cholesteryl and glyceryl ester) and their salt; Two ten diester of thiodipropionic acid; The two octadecyl esters of thiodipropionic acid; Thiodipropionic acid and its derivant (ester for example; Ether; Peptide; Fat; Nucleotide; Nucleoside and salt) and sulphur oxime (sulphoxime) chemical compound of very low tolerance dose (for example pmol to μ mol/kg) (fourth methyllanthionine sulphur oxime for example; Homocysteine sulphur oxime; Fourth methyllanthionine sulfone; Penta; Oneself; Heptan methyllanthionine sulphur oxime); (the alpha-hydroxy fatty acid for example of (metal) chelating agen in addition; Palmic acid; Phytic acid; Lactoferrin); Alpha-hydroxy acid (citric acid for example; Lactic acid; Malic acid); Humic acid; Bile acid; Bile extract; Bilirubin; Biliverdin; EDTA; EGTA and their derivant; Unsaturated fatty acid and their derivant (gamma-Linolenic acid for example; Linoleic acid; Oleic acid); Folic acid and derivant thereof; The alanine acetoacetic acid; Flavonoid; Polyphenol; Catechol; Ubiquinone and pantothenylol and their derivant; Vitamin C and derivant (ascorbyl palmitate for example; The magnesium ascorbate phosphate ester; The ascorbic acid acetas); The coniferyl benzoate of tocopherol and derivant (for example vitamin e acetate) and benzoin resin; Rutinic acid and its derivant; Ferulic acid and its derivant; Butylated hydroxytolyene; Butylated hydroxyanisole; Nor-dihydroguaiaretic acid; Nordihydroguaiaretic acid; Trihydroxybutyrophenone; Uric acid and its derivant; Mannose and its derivant, zinc and its derivant (ZnO for example; ZnSO ₄); Selenium and derivant (for example SLM); Those derivants (for example salt, ester, ether, sugar, nucleotide, nucleoside, peptide and lipid) of stibene and its derivant (for example stibene oxide, trans stibene oxide) and the above-mentioned active component of mentioning that is fit to according to the present invention.

When providing, be used for the compositions that external uses and comprise solvent: water or aqueous solution by following example description as solution, emulsion or dispersion liquid; Oil is capric acid or sad triglyceride, preferably Oleum Ricini for example; Fat, wax and other natural and synthetic lipid, the alcohol of fatty acid and low C number preferably, for example with isopropyl alcohol, the ester of propylene glycol or glycerol; Aliphatic alcohol and low C count alkanoic acid or with the ester of fatty acid (fatty accord); The alcohol of low C number; Glycol or polyalcohols and their ether; Preferably ethanol, isopropyl alcohol, propylene glycol, glycerol, ethylene glycol, ethylene glycol monoethyl ether or ethylene glycol monobutyl ether, propylene glycol monomethyl ether, dihydroxypropane single-ether or propylene glycol monobutyl ether, diethylene glycol monomethyl ether or diethylene glycol monoethyl ether and similar products.And, can use the mixture of the above-mentioned solvent of mentioning.When especially mentioning alcoholic solvent, water can be other composition.

Advantageously be selected from saturated and/or undersaturated, the side chain and/or the unbranched alkanoic acid of chain length according to the oil phase of emulsion of the present invention, oleogel or aqueous dispersions or fat dispersion liquid and have the group that the ester of saturated and/or undersaturated, side chain and/or unbranched alcohol of the chain length of 3 to 30 carbon atoms is formed, be selected from the group that the ester of saturated and/or undersaturated, side chain and/or the unbranched alcohol of aromatic carboxylic acid and the chain length with 3 to 30 carbon atoms is formed with 3 to 30 carbon atoms.In this case; These esters oil can advantageously be selected from the group of being made up of following: isopropyl myristic acid ester; Isopropyl palmitate; Isopropyl stearate; Acid isopropyl; N-butyl stearate; The just own ester of lauric acid; Oleic acid ester in the positive last of the ten Heavenly stems; The different monooctyl ester of stearic acid; Stearic acid ester in the different ninth of the ten Heavenly Stems; Isononyl isononanoate; 2-ethylhexyl cetylate; 2-ethylhexyl laurate; 2-hexyl decyl stearate; 2-octyl group lauryl cetylate; Cetiol; Oleyl erucate; The erucyl alcohol oleate; Synthesizing of erucyl alcohol eruciate and these esters; Semi-synthetic and natural mixture, for example Simmondsia chinensis oil.

And; Oil phase can advantageously be selected from side chain and unbranched Hydrocarbon and Hydrocarbon wax; The group of silicone oil, dialkyl ethers, the group of the group of saturated or undersaturated side chain or unbranched alcohol and fatty acid triglycercide (promptly have 8 to 24 especially saturated and/or unsaturated, the side chain of the chain length of 12-18 C atom and/or the triglyceride of unbranched alkanoic acid).For example, fatty acid triglycercide can advantageously be selected from the for example group of olive oil, Oleum Helianthi, Oleum Glycines, Oleum Arachidis hypogaeae semen, Oleum Brassicae campestris, almond oil, Petiolus Trachycarpi oil, Oleum Cocois, palm-kernel oil and analog of synthetic, semisynthetic and natural oil.Any mixture of these oil and wax component also can advantageously use according to the present invention.If be fit to, use wax, for example cetyl palmitate, be favourable equally as unique lipid component of oil phase.

Oil phase advantageously is selected from the group of being made up of following: 2-ethylhexyl isostearate, octyldodecanol, the different pelargonate of different three decyls, Fancol ID, 2-ethylhexyl cocos nucifera oil acid esters, C12-15-alkyl benzoate, caprylic/capric triglyceride, two caprylyl ethers.Particularly advantageous mixture be C12-15-alkyl benzoate and 2-ethylhexyl isostearate those, those and C12-15 alkylbenzoic acid, 2-ethylhexyl isostearate and the different pelargonate of different three decyls of C12-15 alkyl benzoate and the different pelargonate of different three decyls those.Relevant with Hydrocarbon, liquid paraffin, squalane and Squalene can advantageously use according to the present invention.Oil phase can advantageously comprise ring or linear silicone oils in addition or become by these line of oils fully, but preferably uses the other content of another oil-phase component different with said silicone oil.Cyclomethicone (octamethylcy-clotetrasiloxane) is advantageously used for and remains silicone oil used according to the invention.Yet, can other silicone oil advantageously used according to the invention, for example hexamethyl cyclotrisiloxane, polydimethylsiloxane, gather (methyl phenyl siloxane).And particularly advantageous mixture is those of those and cyclomethicone and 2-ethylhexyl isostearate of cyclomethicone and the different pelargonate of different three decyls.

If be fit to; Water advantageously comprises low C number alone or in combination in each instance the alcohol that contains according to preparation of the present invention; Glycol or how pure and mild their ether; Ethanol preferably; Isopropyl alcohol; Propylene glycol; Glycerol; Ethylene glycol; Ethylene glycol monoethyl ether or ethylene glycol monobutyl ether; Propylene glycol monomethyl ether; Dihydroxypropane single-ether or propylene glycol monobutyl ether; Diethylene glycol monomethyl ether or diethylene glycol monoethyl ether and similar products; The alcohol of low C number in addition; Ethanol for example; Isopropyl alcohol; 1, the 2-propylene glycol; Glycerol and one or more thickening agents especially, it advantageously is selected from by silicon dioxide; Aluminium silicate; Polysaccharide and their derivant be hyaluronic acid for example; Xanthan gun; The group that hydroxypropyl emthylcellulose is formed; Be selected from the group of polyacrylate advantageous particularly, preferably be selected from so-called Carbopol for example 980; 981; 1382; The polyacrylate of the group that 2984 and 5984 type Carbopol form.

Gel used according to the invention comprises the alcohol of low C number usually; For example ethanol, isopropyl alcohol, 1; 2-propylene glycol, G & W; Or the oil of above mentioning under the situation that thickening agent exists; Its preferably silicon dioxide or aluminium silicate in the instance of oil-containing-spirituosity gel, and in the instance of moisture-spirituosity or alcoholic gel its polyacrylate preferably.

Solid bar comprises for example natural or synthetic wax, aliphatic alcohol or fatty acid ester.The base substance that is suitable for use as according to the routine of cosmetic stick of the present invention is liquid oil (for example liquid paraffin, Oleum Ricini, isopropyl myristic acid ester), semi-solid composition (for example vaseline oil, lanoline), solid composition (for example Apis cerana Fabricius, ceresine and microwax or paraffin (ozocerite)) and dystectic wax (for example carnauba wax, candelilla wax).

Be used for according to cosmetics of the present invention and/or dermatological preparation can be the propellant Hydrocarbon (propane, butane, isobutene .) for example of conventional known volatile, liquefaction from the propellant that is fit to of aerosol container ejection, it can be separately or as each other mixture use.Air pressurized also can advantageously be used.Certainly, those of skill in the art are familiar with having the fact that is suitable for being committed to the form of aerosol preparations the nontoxic propellant of practice of the present invention in principle; Yet, because their situation subsidiary to the unacceptable influence of environment or other, suggestion without these-particularly fluorohydrocarbon and fluoro chlorohydrocarbon (FCHC)-processing.

Being used for the compositions that external according to the present invention uses also can be the form of gel, its not only comprise effective dose according to active component of the present invention and the conventional thus solvent that uses, also comprise organic thickening agent.The instance of these thickening agents comprises arabic gum, Xanthan gun, sodium alginate, cellulose derivative; Preferably methylcellulose, hydroxy methocel, hydroxyethyl-cellulose, hydroxypropyl cellulose, hydroxypropyl emthylcellulose or inorganic thickening agent, for example aluminium silicate is for example such as the mixture of bentonite or Polyethylene Glycol and polyglycol distearate or polyglycol distearate.

Comprising above, the instance of the acceptable cosmetics/dermatological carrier formulation of the active component of record can comprise following ingredients: Xanthan gun, glycerin 99.7%, sequestrene Na4, tristerin and PEG-100 stearate (ARLACEL ^TM165), cetyl alcohol, isopropyl palmitate, BHT (BHT), methyl parahydroxybenzoate, propylparaben and deionized water.Comprising above, another instance of the acceptable cosmetics/dermatological carrier formulation of the active component of record can comprise following inert fraction: stearic acid, cetyl alcohol, laureth 9 (Laureth) 4, CARSONOL ^TMSles, propyl p-hydroxybenzoate, ascorbyl palmitate, propylene glycol, CARBOPOL ^TM974P, methyl hydroxybenzoate, KOH (10%) and H ₂O.

Above Ji Lu compositions can prepare through process for example as hereinafter:

1. make up and the thawing oil phase: stearic acid, cetyl alcohol, laureth4, propyl p-hydroxybenzoate and ascorbyl palmitate;

2. in glass beaker, with propylene glycol and water combination, with high-speed screw dispersed with stirring methyl hydroxybenzoate and CARBOPOL ^TM

3. with CARSONOL ^TMSles adds to the product of step (2);

4. the product to 65 of heating steps (3) is ℃-70 ℃;

5. under mixing, the product of step (1) is added to the product of step (4) and mix;

6. mixture is cooled to 40 ℃;

7. it is good to add solvent part and manual mixing;

8. add KOH solution with neutralization; And

9. lucifuge.

General introduction

In each instance; Compositions can compatibly contain one or more other activating agent; It can be selected from A/B-cis spirostane or spirostene steroid sapogenines and its ester, ether, ketone and glycosylated form, other Saponin (unit), other non-Saponin (unit) activating agent or its any combination.Compositions can contain one or more biological inert compositions; For example diluent, carrier and excipient; Its for the physiology go up effective composition preparation, use or send relevant purpose and serve, or the benefit of the associating different with the physiological role of active ingredient is provided to the curee.Carrier can comprise for example soya bean protein of vegetable material.Depend on the mode used and the character of dosage form, compositions can for example also comprise any or multiple in preservative agent, filler, disintegrating agent, humidizer, emulsifying agent, suspending agent, sweeting agent, flavoring agent, aromatic, antibacterial agent, antifungal, lubricant and the dispersant.

The compositions that is used for using in the present invention, especially pharmaceutical composition can be unit dosage forms, and these dosage forms of a certain quantity can be applied to the curee in the section sometime according to the patient's condition that pending or prevention are arranged thus.Selectively, compositions can be a bulk form, thus according to having the patient's condition pending or prevention to measure the bulk composition of a certain weight or volume and being applied to the curee in the section sometime.

Yet, do not think that toxicity is the problem of these activating agents, even at higher dosage.In the selection of suitable dosage so those skilled in the art's in the art the ability, and there is not over-drastic burden.The about 0.3mg/kg body weight of the dosage of the activating agent of being used, preferably once-a-day administration.More generally, dosage will about 0.1 and about 25mg/kg between, for example about 1 and about 10mg/kg between, preferably use once or twice every day.For human use of growing up, dosage can be conventional about 10 and about 700mg between every day.

The compositions that is used for using in the present invention can compatibly comprise other treatment and/or non-treatment bioactivator, and is as discussed above.

The compositions that is used for using in the present invention can be a unit dosage forms, and thus, these dosage forms of a certain quantity can be applied to the curee in the section sometime according to the patient's condition that pending or prevention are arranged.Selectively, compositions can be a bulk form, thus according to having the patient's condition pending or prevention to measure the bulk composition of a certain weight or volume and being applied to the curee in the section sometime.

The needed dosage of activating agent will depend on that the severity of the symptom that pending or prevention are arranged changes to a great extent.The concentration that flies in mole to the micro-molar range is effectively, and for example about 1fM is to about 5 μ M.The experimental work of report shows the external EC of the smilagenin of anti-nervous lesion in the culture among the embodiment 12 ₅₀Be 13.4fM.Usually; Plasma concentration is normally preferred in the body in picomole to the micro-molar range (for example at nanomole to micro-molar range); For example be higher than about 1pM, for example at about 1pM to about 5 μ M, for example about 1pM is to about 3 μ M, extremely about 700nM, about 0.1nM extremely in the scope of about 500nM for example of about 10pM for example.Below picomole, the activity in vivo of activating agent often reduces.More than the micromole, the curee will simply mean activating agent to self regulating of overtreatment with relevant resistance and be wasted.Yet, as among the application embodiment showed, do not think that toxicity is the problem of these activating agents, even at higher dosage.In the selection of the dosage that is fit to so those skilled in the art's in the art the ability, and there is not over-drastic burden.The application dosage of activating agent can be for example to be higher than about 0.1mg/kg body weight, for example is higher than about 0.3mg/kg body weight, preferably uses once every day.More generally, dosage about 0.1 and about 25mg/kg between, for example about 1 and about 10mg/kg between, preferably use once every day.For human use of growing up, dosage is conventionally about 10 and about 700mg between every day.

For the composition forms that is fit to and dosage and according to the present invention the further details of the instance of the accessible patient's condition and disease, please refer to WO-A-99/48482, WO-A-99/48507, WO-A-01/23407, WO-A-01/23408, WO-A-02/079221, WO-A-03/082893, WO-A-2005/105825 and WO-A-2006/048665.

Activating agent is adapted at preparing with one or more carriers, excipient and/or diluent in the compositions.Generally speaking, can use to be used for pharmaceutical composition, Orally administered composition is food, food supplement and beverage for example, or topical composition for example any conventional carrier, excipient and/or the diluent of cosmetics, eye or skin preparation.

Many in the activating agent be lipophilic relatively and in this instance dissolving and/or suspension and/or dispersant applicable to keeping dissolving or suspension or the dispersion of activating agent in compositions.

Two groups of dissolvings can mentioning especially and/or suspension and/or dispersant are MCT and medium-chain fatty acid (MCFA).These be have chain length about 4 and about 12 carbon atoms between the lipophilic compound of fatty acid chain.

The preferred examples of MCT is represented by following general formula (I):

Wherein, Ra, Rb and Rc are independently from each other the saturated or unsaturated fatty acid residue that in carbon backbone chain, has 4 to 12 carbon atoms.

The preferred examples of MCFA is represented by following general formula (II):

HO-CO-Rd

(Ⅱ)

Wherein Rd is the saturated or unsaturated fatty acid residue that in carbon backbone chain, has 4 to 12 carbon atoms.

The instance of Ra, Rb, Rc and Rd comprises the residue of caproic acid (C6:0), sad (C8:0), capric acid (C10:0) and lauric acid (C12:0).In the nomenclature of standard, the length of the numeral carbochain after the letter C and the number of colon (:) numeral unsaturated bond afterwards.These MCT and MCFA can for example Oleum Cocois, palm-kernel oil and Camphora drupe (fruit) obtain from natural origin in known manner.Can exist a kind of in commercially available MCT or the MCFA product or more than a kind of residue of fatty acid.

The MCT that is used for using in the present invention can for example be selected from three-C6:0MCT, three-C8:0MCT and three-C10:0MCT.

Industrial usability and function

The present invention makes the disease of NF mediation in human and the non-human mammal and the therapeutic of function and the method for self regulating that non-therapeutic is handled get first; Wherein physiological responses in the scope relatively widely of the dosage of activating agent be not dose dependent but self regulate, do not have disadvantageous side effect with expected useful physiological role simultaneously or toxic mode provides narrow relatively " treatment window ".Therefore said processing can tolerate overtreatment in the scope relatively widely.This character make said processing be suitable for self using or clinical scenarios outside other situation, this is that neurological is handled a kind of characteristics that can't realize so far with other.Said dose is the true potential function of the present invention outside clinical scenarios of further supporting of micromolecule rather than peptide (for example albumen), and wherein being used for the peptide activating agent maybe non-availability to the elaborate delivery apparatus of directly using of brain or CNS.

Can be old relatively or be in bad relatively general health that they usually are prone to suffer from some other diseases of these minutes apoplexy due to endogenous wind owing to suffer from many patients of neurological, psychiatry, inflammatory, allergia, immunity or tumprigenicity disease.Which usually can't will occur with in any definitiveness many other diseases of expection or the patient's condition.Before the present invention, these other the disease or the patient's condition or individual liability to them make the taboo that is treated as of protopathy disease, because very frequent processing will have the material risk that promotes these other diseases among such patient or the patient's condition.Therefore, the present invention with than before be more prone to represent these healthy key areas Chinese medicine of human and animal and the health care important advance aspect putting into practice with simple and the patient's that can be applied to organize widely the effectiveness of mode in handling the disease and the patient's condition.

The accompanying drawing summary

For further exemplary illustration, will describe now and support data of the present invention, it only is exemplary and nonrestrictive.

In the accompanying drawing of enclosing:

Fig. 1 shows that neuronic smilagenin pretreatment is to the anti-MPP of neuroprotective unit ⁺The effect of inductive infringement;

Fig. 2 shows that chinaroot greenbrier sapogenin is to (a) compound muscle action potential (CMAP) amplitude in carrying out property motor neuron (pmn) mice, and (b) grid is tested and (c) influence of survival rate; With

Fig. 3 shows that chinaroot greenbrier sapogenin, smilagenin and 4-methyl catechol are in time to CMAP (a) amplitude, (b) incubation period and (c) influence of persistent period in the harmful mice of nerve damage.

Embodiment and detailed description of the drawings

In the following example and detailed description of the drawings, used following abbreviation: h=hour; Min=minute; S=second; S.c=is subcutaneous, and p.o=is oral.The percentage ratio of the composition of the compositions of solid bag solid (solid in solid) or liquid bag solid (solid in liquid) or solid bag liquid (liquid in solid) is the weight meter.The percentage ratio of the composition of the compositions of liquid bag liquid (liquid in liquid) is stereometer.

Embodiment 1

Chinaroot greenbrier sapogenin and smilagenin do not combine with several enzymes and receptor

The research chinaroot greenbrier sapogenin combines receptor listed in the active influence of enzyme listed in the table 1 hereinafter and chinaroot greenbrier sapogenin and the table 1 hereinafter.

Use following method research enzymatic activity to regulate: chinaroot greenbrier sapogenin is added that with every kind of enzyme the specific substrate of every kind of enzyme hatches.Reaction is stopped after incubation period, and measure the minimizing of specific substrate under the situation that chinaroot greenbrier sapogenin do not exist and exists or the increase of specificity product, and calculate the percentage ratio inhibition of reacting under the situation that chinaroot greenbrier sapogenin exists.The amount of employed enzyme, incubation conditions, employed substrate and quantitative methods depend on every kind of particular assay and change.

Use following method investigation receptors bind: with chinaroot greenbrier sapogenin with express interested receptor tissue or cell homogenates and have to the radiolabeled chemical compound of concentration known of affinity of interested receptor hatch.To there be bonded radiolabeled chemical compound to remove and the quantitative bonded amount of specificity after incubation period.The relatively bonded amount of specificity under chinaroot greenbrier sapogenin existence and the non-existent situation, and calculating chinaroot greenbrier sapogenin suppresses the bonded percentage ratio of radiolabeled chemical compound.The source of receptor, incubation conditions, employed radiolabeled chemical compound depend on every kind of particular assay and change.

The result is shown in hereinafter in the table 1.

Table 1 chinaroot greenbrier sapogenin is to the influence of enzyme and receptors bind mensuration

NS=does not have remarkable response.Significance is adopted as＞30% stimulation or inhibition.

Use and above describe the same method, study receptor listed in smilagenin (1 μ M) and the table 2 hereinafter combine and smilagenin to the active influence of enzyme listed in the table 2 hereinafter.

Table 2 smilagenin is to the influence of receptors bind mensuration and enzyme

Target	Species	Influence (%)
			In conjunction with measuring
Adensonine?A 1	Human	NS
			Adensonine?A 2A	Human	NS
Adensonine?A 3	Human	NS
			Adrenergic α 1A	Rat	NS
Adrenergic α 1B	Rat	NS
			Adrenergic α 1D	Human	NS
Adrenergic α 2A	Human	NS
			Adrenergic α 2C	Human	NS
Alpha 1 beta-adrenergic 1	Human	NS
			Alpha 1 beta-adrenergic 2	Human	NS
Target	Species	Influence (%)
			Alpha 1 beta-adrenergic 3	Human	NS
Adrenomedullin AM 1	Human	NS
			Adrenomedullin AM 2	Human	NS
Aldosterone	Rat	NS
			C5a anaphylatoxin	Human	NS
Androgen (testosterone) AR	Rat	NS
			Angiotensin AT 1	Human	NS
Angiotensin AT 2	Human	NS
			APJ	Human	NS
Atrial natriuretic factor	Cavia porcellus	NS
			Bombesin BB1	Human	NS
Bombesin BB2	Human	NS
			Bombesin BB3	Human	NS
Kallidin I B 1	Human	NS
			Kallidin I B 2	Human	NS
Calcitonin	Human	NS
			Calcitonin-gene-related peptide CGRP 1	Human	NS
Calcium channel L-type, benzothiazepine	Rat	NS
			Calcium channel L-type, dihydropyridine	Rat	NS
Calcium channel L-type, phenylalkylamine	Rat	NS
			Calcium channel N-type	Rat	NS
Cannabinoid CB 1	Human	NS
			Cannabinoid CB 2	Human	NS
Target	Species	Influence (%)
			Chemotactic factor CCR 1	Human	NS
Chemotactic factor CCR2B	Human	NS
			Chemotactic factor CCR 4	Human	NS
Chemotactic factor CCR 5	Human	NS
			Chemotactic factor CXCR 1	Human	NS
Chemotactic factor CXCR 1(IL-8R B)	Human	NS
			Cholecystokinin CCK 1(CCK A)	Human	NS
Cholecystokinin CCK 2(CCK B)	Human	NS
			Colchicine		NS
Corticotropin-releasing factor (CRF 1)	Human	NS
			Dopamine D 1	Human	NS
Dopamine D 2S	Human	NS
			Dopamine D 3	Human	NS
Dopamine D 42	Human	NS
			Dopamine D 5	Human	NS
Endothelin ET A	Human	NS
			Endothelin ET B	Human	NS
Epidermal growth factor (EGF)	Human	NS
			Erythropoietin EPOR	Human	NS
Estrogen (ER α)	Human	NS
			Estrogen (Er β)	Human	NS
G protein coupled receptor GPR103	Human	NS
			G protein coupled receptor GPR8	Human	NS
Target	Species	Influence (%)
			GABA AChloride channel, TBOB	Rat	NS
GABA AFlunitrazepam, maincenter	Rat	NS
			GABA A5-aminomethyl-3-hydroxyisoxazole, maincenter,	Rat	NS
GABA B1A	Human	NS
			GABA B1B	Human	NS
Gabapentin	Rat	NS
			Galanin GAL1	Human	NS
Galanin GAL2	Human	NS
			Glutamate, Glu, AMPA	Rat	NS
Glutamate, Glu, kainate	Rat	NS
			Glutamate, Glu, NMDA, excitement	Rat	NS
Glutamate, Glu, NMDA, glycine	Rat	NS
			Glutamate, Glu, NMDA, phencyclidine	Rat	NS
Glutamate, Glu, NMDA, polyamines	Rat	NS
			Glycine, strychnine-sensitivity	Rat	NS
Growth hormone secretagogues (GHS, Ghrelin)	Human	NS
			Histamine H 1	Human	NS
Histamine H 2	Human	NS
			Histamine H 3	Human	NS
Histamine H 4	Human	NS
			Imidazoline I 2, maincenter	Rat	NS
Inositoltriphosphoric acid IP 3	Rat	NS
			Insulin	Rat	NS
Target	Species	Influence (%)
			Interleukin I L-1	Mice	NS
Interleukin I L-2	Mice	NS
			Interleukin I L-6	Human	NS
Leptin	Mice	NS
			Leukotriene, BLT (LTB 4)	Human	NS
Leukotriene, cysteinyl CysLT 1	Human	NS
			Leukotriene, cysteinyl CysLT 2	Human	NS
Melanocortin MC 1	Human	NS
			Melanocortin MC 3	Human	NS
Melanocortin MC 4	Human	NS
			Melanocortin MC 5	Human	NS
Melatonin MT 1	Human	NS
			Melatonin MT 2	Human	NS
Motilin	Human	NS
			Muscarine M 1	Human	NS
Muscarine M 2	Human	NS
			Muscarine M 3	Human	NS
Muscarine M 4	Human	NS
			Muscarine M 5	Human	NS
N-formyl peptide receptor FPR1	Human	NS
			N-formyl peptide receptor appearance FPRL1	Human	NS
Neuromedin U MNU1	Human	NS
			Neuromedin U MNU2	Human	NS
Target	Species	Influence (%)
			Neuropeptide tyrosine Y 1	Human	NS
Neuropeptide tyrosine Y 2	Human	NS
			Neurotensin NT 1	Human	NS
Nicotinic acetycholine	Human	NS
			Nicotinic acetycholine α 1, bungarotoxin	Human	NS
Nicotinic acetycholine α 7, bungarotoxin	Rat	NS
			Opium δ (OP1, DOP)	Human	NS
Opium κ (OP2, KOP)	Human	NS
			Opium μ (OP3, MOP)	Human	NS
Nociceptin ORL1	Human	NS
			Buddhist ripple ester	Mice	NS
Platelet activating factor (PAF)	Human	NS
			Platelet-derived somatomedin (PDGF)	Mice	NS
Potassium channel [K A]	Rat	NS
			Potassium channel [K ATP]		NS
Potassium channel [SK CA]	Rat	NS
			Potassium channel HERG	Human	NS
Progesterone PR-B	Human	NS
			Prostanoid CRTH2	Human	NS
Prostanoid DP	Human	NS
			Prostanoid EP 2	Human	NS
Prostanoid EP 4	Human	NS
			The prostanoid thromboxane A 2(TP)	Human	NS
Target	Species	Influence (%)
			Purine can P 2X		NS
Purine can P 2Y	Rat	NS
			Tetanus appearance X receptor RXR α	Human	NS
The rolipram receptor	Rat	NS
			Li Luoding RyR3	Rat	NS
Serotonine, 5-HT 1A	Human	NS
			Serotonine, 5-HT 1B	Rat	NS
Serotonine, 5-HT 2B	Human	NS
			Serotonine, 5-HT 2C	Human	NS
Serotonine, 5-HT 3	Human	NS
			Serotonine, 5-HT 4	Cavia porcellus	NS
Serotonine, 5-HT 5A	Human	NS
			Serotonine, 5-HT 6	Human	NS
σ(sigma)σ1		NS
			σ(sigma)σ2	Rat	NS
The sodium channel receptor, site 2	Rat	NS
			Somat sst1	Human	NS
Somat sst2	Human	NS
			Somat sst3	Human	NS
Somat sst4	Human	NS
			Somat sst5	Human	NS
Tachykinin NK-1 1	Human	NS
			Tachykinin NK-1 2	Human	NS
Target	Species	Influence (%)
			Tachykinin NK-1 3	Human	NS
Thyroxin	Rat	NS
			Throtropin releasing hormone (TRH)	Rat	NS
Transforming growth factor-beta (TGF-β)	Mice	NS
			Carrier, vidarabine	Cavia porcellus	NS
Carrier, choline	Rat	NS
			Carrier, dopamine (DAT)	Human	NS
Carrier, GABA	Rat	NS
			Carrier, monoamine	Rabbit	NS
Carrier, norepinephrine carrier (NET)	Human	NS
			Carrier, serotonine (SERT)	Human	NS
Tumor necrosis factor (TNF), non-selective	Human	NS
			Urotensin II	Human	NS
Rhizoma et radix valerianae is plain	Rat	NS
			VEGF (VEGF)	Human	NS
Vasoactive intestinal peptide, VIP 1	Human	NS
			Vassopressin V 1A	Human	NS
Vassopressin V 1B	Human	NS
			Vassopressin V 2	Human	NS
Vitamin D 3	Human	NS
			Functional examination		NS
The albumen serine/threonine kinase, AKT1 (PRKBA)	Human	NS
			The albumen serine/threonine kinase, AKT3 (PRKBG)	Human	NS

NS=does not have remarkable response.Significance is adopted as >=30% stimulation or inhibition.

The activity that chinaroot greenbrier sapogenin and smilagenin and many receptors do not combine and do not regulate many enzymes.Because known these receptors and enzyme are participated in nerve, consciousness and motor path; Infer in the scope of the knowledge that these experiments obtain, the activity that chinaroot greenbrier sapogenin and smilagenin be anti-to have the patient's condition and a disease in nerve, consciousness and motion source is not via receptors bind or enzyme adjusting and take place.

Embodiment 2

Chinaroot greenbrier sapogenin and smilagenin be instantaneous increasing in the neuron of cultivating under the external primary condition Add neurotrophic factor mRNA

Use special culture medium and condition, the neuron of fresh separated can In vitro culture; External environment is different with physiological environment, cause neuron be more stress and damaged by neuron.The level of neuron infringement will depend on employed accurate condition and between culture medium and culture medium, change.Can be after the level of neuron infringement through adding pathology agent (for example amyloid beta or MPP ⁺) and significantly increase.

Through the method culture of rat cortical neuron of describing before revising (Singer waits the people, Neuroscience Letters, 1996,212,13-16 page or leaf).Cultivated beginning 12 days afterwards; Add the derivant 4-methyl catechol (0.5mM) that chinaroot greenbrier sapogenin (30nM), smilagenin (30nM), NGF and BDNF discharge (people such as Saporito, Experimental Neurology., 1993; 123, the 295-302 pages or leaves; People such as Nitta, Journal of Pharmacology and Experimental Therapeutics, 1999,291,1276-1283 page or leaf) or vehicle (dimethyl sulfoxide, DMSO, 0.25%) 1,3 or 6h.After hatching, use the quantitatively total Messenger RNA (mRNA) of real-time reverse transcriptase polymerase chain reaction (rtRT-PCR).

The result is shown in hereinafter in the table 3.

Table 31,3 and 6h hatch the influence that back chinaroot greenbrier sapogenin, smilagenin and 4-methyl catechol are expressed BDNF in the rat cortical neuron and trkB mRNA

Chinaroot greenbrier sapogenin and smilagenin all instantaneous in the cortical neuron of fresh separated (behind the 3h) increase the level of the mRNA of BDNF and BDNF receptor trk-B (tyrosine receptor kinase neurotrophin receptor).

In different experiments, through the method culture of rat cortical neuron of describing before revising (Eckenstein and Sofroniew, Journal of Neuroscience, 1983,3,2286-2291 page or leaf).At the 8th day, culture medium is changed to the culture medium 48 hours that contains vehicle (DMSO, 0.5%) or smilagenin (10 μ M) and passes through the level that rt RT-PCR measures the BDNF mRNA in the cortical neuron.

The result is shown in hereinafter in the table 4.

Table 4 uses chinaroot greenbrier sapogenin to hatch the influence of BDNF mRNA in the rat cortical neuron being expressed in 48 hours

Condition	The relative quantity of BDNF mRNA (% of contrast)
		Contrast (DMSO, 0.5%)	?100.0±0.0
Smilagenin (10 μ M)	?103.9±6.5

Meansigma methods ± standard error of mean; N=3.

Hatch the level of BDNF mRNA in the cortical neuron that did not increase fresh separated in 48 hours with smilagenin (10 μ M).The data consistent that shows in this and the table 3 shows that smilagenin and chinaroot greenbrier sapogenin are at 3h rather than 6h increase BDNF mRNA.In addition, the snap (table 3) of smilagenin is not overcome by the smilagenin (table 4) of high concentration.

Embodiment 3

Smilagenin causes neurotrophy in the neuron of the external cultivation that is exposed to the pathology agent Factor mRNA expresses significantly to be increased

Smilagenin increases the BDNFmRNA in the cortical neuron that is exposed to amyloid-beta before.

Through the method culture of rat cortical neuron of describing before revising (Eckenstein and Sofroniew, Journal of Neuroscience, 1983,3,2286-2291 page or leaf).At the 8th day, culture medium is changed to the culture medium that contains vehicle (DMSO, 0.5%) or smilagenin (10 μ M).At the 10th day, the primary cortical neuron of rat was exposed to amyloid-beta nearly 48 hours and in the level of next measuring the BDNF mRNA in the cortical neuron in 48 hours through rt RT-PCR in 37 ℃.

The result is shown in hereinafter in the table 5.

Table 5 increases the BDNF mRNA in the rat cortical neuron with being exposed to amyloid-beta after the smilagenin preincubate 48h

Meansigma methods ± standard error of mean; N=3, ^*=p＜0.01, ^*=p＜0.05 is compared with the corresponding time point of independent amyloid-beta.Through student t check carrying out statistical analysis.

Be exposed to smilagenin preincubate 48h and amyloid-beta afterwards and produce remarkable and lasting increase in the expression of the BDNF mRNA in the rat cortical neuron.

Before increasing, smilagenin is exposed to MPP ⁺Dopaminergic neuron in GDNF mRNA

Use revise a little before the method described prepare rat dopaminergic neuron people such as (, Journal of Neuroscience, 1992,12, the 1409-1415 page or leaf) Brouard.At culture medium MPP ⁺After 5 days, added special dopaminergic nerve toxin or vehicle (saline) 48 hours in (2 μ M).Afterwards culture medium is replaced with the fresh culture that contains smilagenin (10 μ M) or vehicle (DMSO, 0.5%), and after 10min and 2,24,48h, pass through the level that rt RT-PCR measures the GDNF mRNA in the dopaminergic neuron.

The result is shown in hereinafter in the table 6.

The increase of table 6 smilagenin is exposed to MPP ⁺GDNF mRNA in the back rat dopaminergic neuron expresses

Meansigma methods ± standard error of mean; N=3, * *=p＜0.01, *=p＜0.05 is compared with contrast.Carry out the statistical analysis of the GDNF mRNA between each time point contrast and the smilagenin through student t check.

Be exposed to MPP ⁺The remarkable increase that the back uses the smilagenin processing to cause the GDNF mRNA in the rat dopaminergic neuron to express.When being increased in 24h maximum and afterwards 48 with 72h after reduction.

Embodiment 2 and 3 proof smilagenins and chinaroot greenbrier sapogenin increase neurotrophic factor mRNA expresses.And the influence that neurotrophic factor mRNA is expressed of smilagenin and chinaroot greenbrier sapogenin is depended on neuronic situation and on its degree (persistent period and intensity), is changed to some extent.In the neuron that is to cultivate under the basic condition; Smilagenin and chinaroot greenbrier sapogenin produce the instantaneous increase (nearly 140% of contrast) of neurotrophic factor rna level, and it is observed when 3h (table 3) but does not observe when 6h (table 3) or during 48h (table 4).On the contrary, be exposed to pathology agent (for example amyloid-beta or MPP ⁺) the neuron of cultivation in, smilagenin produce neurotrophic factor mRNA express with (table 5 and table 6) significantly (up to contrast 3319%) and the increase of last much longer (reaching 72 hours).Result proof depends on that the neurotrophy inducer effect self of degree smilagenin and chinaroot greenbrier sapogenin of the infringement of system regulates that they are own, and promptly smilagenin and chinaroot greenbrier sapogenin be not with self regulatory mechanism interruption of neurotrophic factor or transship.

Embodiment 4

Smilagenin does not change neural battalion in the neuron of external cultivation under primary condition Support the expression of factor protein

Through the method culture of rat cortical neuron of describing before revising (Eckenstein and Sofroniew, Journal of Neuroscience, 1983,3,2286-2291 page or leaf).At the 8th day, culture medium is changed to the culture medium that contains vehicle (DMSO, 0.5%) or smilagenin (10 μ M).At the 12nd day, measure the concentration of BDNF in the culture medium.

The result is shown in hereinafter in the table 7.

Table 7 is hatched the bdnf protein level that does not change in the neuron of cultivating under the external primary condition with smilagenin

Condition	BDNF concentration (pg/ml)
		Contrast (DMSO, 0.5%)	3.66±0.05
Contrast+smilagenin (10 μ M)	3.67±0.05

Meansigma methods ± standard error of mean; N=6

Smilagenin does not increase the BDNF level in the neuron of cultivating under the external primary condition.

Embodiment 5

Smilagenin increases neurotrophy in the neuron of the external cultivation that is exposed to the pathology agent The expression of factor protein

Chinaroot greenbrier sapogenin and smilagenin before be exposed to and increase bdnf protein in the cortical neuron of amyloid-beta and increase neuronal survival and neurite outgrowth

Through the method culture of rat cortical neuron of describing before revising (Eckenstein and Sofroniew, Journal of Neuroscience, 1983,3,2286-2291 page or leaf).At the 8th day, culture medium is changed to the culture medium that contains vehicle (DMSO, 0.5%) or smilagenin or chinaroot greenbrier sapogenin (10 μ M).At the 10th day, the primary cortical neuron of rat is exposed to amyloid-beta (10 μ g/ml) 48h and measures the concentration of BDNF in the culture medium, the number and the neurite outgrowth (only smilagenin) of CAT (ChAT) positive cell in 37 ℃.

The result is shown in hereinafter in the table 8.

Being exposed to amyloid-beta after table 8 external use smilagenin or the chinaroot greenbrier sapogenin preincubate 48h increases the bdnf protein level and prevents neuron infringement and neuronatrophy

N.m.=does not measure; Meansigma methods ± standard error of mean; N=4-8, ^{++ ++}=p＜0.001 is compared with contrast, ^{* * *}=p＜0.001, ^*=p＜0.005 is compared with independent amyloid-beta.Use one way analysis of variance (ANOVA) and Fisher afterwards to check afterwards and carry out statistical analysis.

Chinaroot greenbrier sapogenin and smilagenin increase BDNF horizontal exceeding control level and the inductive neuron infringement of prevention amyloid-beta in cortical neuron.

Smilagenin before be exposed to MPP ⁺Dopaminergic neuron in increase gdnf protein and increase neuronal survival and neurite outgrowth

Use revise a little before the method described prepare rat dopaminergic neuron people such as (, Journal of Neuroscience, 1992,12, the 1409-1415 page or leaf) Brouard.At the 6th day, with culture medium with fresh culture or contain the fresh culture replacement of smilagenin (10 μ M) or vehicle (DMSO, 0.25%).At the 8th day, add MPP ⁺Concentration, neuron infringement and the neurite outgrowth of (2 μ M) or vehicle (saline) and 48 hours GDNF in the culture medium that afterwards dopaminergic neuron dyeed and measure.

The result is shown in hereinafter in the table 9.

Table 9 smilagenin increases amount and the prevention MPP of GDNF in the culture medium ⁺Neuron infringement and neuronatrophy in the rat dopaminergic neuron after exposing

N.m.=does not measure; Meansigma methods ± standard error of mean; N=5-6, ^{++ ++}=p＜0.001 is compared with contrast, ^*=p＜0.05 and independent MPP ⁺Compare.The number that uses one way analysis of variance and Fisher afterwards afterwards to check to carry out the TH positive neuron and the statistical analysis of neurite outgrowth.Check the statistical analysis that carries out GDNF concentration through student t.

Smilagenin increases amount and the prevention MPP of GDNF in the dopaminergic neuron ⁺Inductive neuron infringement.

The data show smilagenin that shows among the embodiment 4 does not increase neurotrophic factor protein expression in the neuron of the cultivation under the external primary condition.On the contrary, embodiment 5 shows that chinaroot greenbrier sapogenins and smilagenin all increase neurotrophic factor protein expression in the neuron of the external cultivation that is exposed to the pathology agent.Therefore; Chinaroot greenbrier sapogenin is similar to the influence of neurotrophic factor mRNA with them to the proteic influence of neurotrophic factor with smilagenin; Be that chinaroot greenbrier sapogenin and smilagenin do not interrupt self regulatory mechanism of neurotrophic factor or overload, but in fact depend on neuronic demand and stand them.

Because known BDNF, trk-B and GDNF participate in nerve, consciousness and motor path; Infer in the scope of the knowledge that these experiments obtain, the activity that chinaroot greenbrier sapogenin and smilagenin be anti-to have the patient's condition and a disease in nerve, consciousness and motion source comprises the gene expression of enhancing neurotrophic factor and their receptor.

Embodiment 6

Chinaroot greenbrier sapogenin and smilagenin are repaired the BDNF concentration in the older animals

With the Orally administered chinaroot greenbrier sapogenin of old Sprague Dawley (SD) rat (20 months big) or smilagenin (18mg/kg/ days) 3 months.Comparing BDNF with the brain of the rat of youth significantly reduces in old rat brain.The SD rat (4 months big) of youth is used as healthy positive control.When the end of handling, brain is shifted out so that use the quantitative BDNF of ELISA.

The result is shown in hereinafter in the table 11.

Table 11 chinaroot greenbrier sapogenin and smilagenin reverse the decline of the BDNF level in the old rat and with the BDNF level to young state reparation

Condition	BDNF (ng/g tissue)
		Young	1.65±0.09
Old	1.21±0.07 ++++
		Old+chinaroot greenbrier sapogenin (18mg/kg/ days)	1.41±0.07*
Old+smilagenin (18mg/kg/ days)	1.34±0.07*

Meansigma methods ± standard error of mean; N=9-10 uses paired single tail student t check carrying out statistical analysis. ^{++ ++}=p＜0.001 is compared with young rats, ^*=p＜0.05 is compared with old rat.

To the Orally administered chinaroot greenbrier sapogenin of old rat or smilagenin 3 months, decline observed level in the young healthy rat of the BDNF of older animals is reversed, that is, compare with old control rats, said dose significantly increases the BDNF level.

These data show that the said dose of influence that BDNF is expressed is the normalization effect under chronic administration, that is, have through recovery being limited to animal that normal condition protection roughly is processed and avoid the long-term adjusting influence to said dose excessive exposure.

This embodiment has replenished the experiment among the embodiment 9 of PCT patent application WO-A-03/082893 number of incorporating this paper by reference into.That experiment proof smilagenin or chinaroot greenbrier sapogenin significantly reduce or reverse the minimizing of the age is relevant in the rat BDNF, dopamine receptor and muscarinic acetylcholinergic receptor.

Embodiment 7

BDNF and GDNF concentration in the striatum of the mice of smilagenin increase MPTP-infringement

Seven weeks, big male C57bl/6RJ mice (C57 mice) was accepted daily injection 1-methyl-4-phenyl-1; 2; 3; 6-tetrahydropyridine (MPTP; 25mg/kg/ days, lumbar injection, continuous 5 days) and Orally administered smilagenin (10mg/kg/ days) or vehicle (hydroxyl third methylcellulose; The HPMC 0.5%w/v that contains tween 80 0.2%v/v) 60 days, the brain with them shifted out striatum level and the use [I that is used to use quantitative GDNF of ELISA and BDNF afterwards ¹²⁵]-RTI combines quantitative dopamine carrier (DAT) level.DAT is the sign that is used for the neuron infringement of dopaminergic neuron.

By neurotoxin MPP ⁺, the for example degeneration (people such as Mytinlineou, Science, 225,529-531 (1984)) of observed nigrostriatal dopaminergic neuron in the parkinson of the metabolite of MPTP, the infringement analog neuron retrograde affection that causes.Change the level that comprises the increase of dopamine and its metabolite in black substance compact part and the caudatum (people such as Burns by the most significant biochemistry of this toxin-induced; Proc.Natl.Acad.Sci.USA; 80; 4546-4550 (1983)) minimizing that dopamine is taken in and in the nigrostriatal synaptosome goods (people such as Heikkila; J.Neurochem.; 44,310-313 (1985)).

Therefore the mice that employed MPTP handles in this experiment provides the acceptable model of parkinson and similar motion-sensory nerve degeneration patient's condition.

The result is shown in hereinafter in the table 12 and 13.

Striatum GDNF and BDNF in the mice of table 12 smilagenin increase MPTP infringement

Meansigma methods ± standard error of mean; N=8-11 ^*=p＜0.01 is compared with the mice of MPTP infringement.Use one way analysis of variance and Tukey afterwards multiple comparisons check carrying out statistical analysis afterwards.

Striatum DAT level in the mice of table 13 smilagenin increase MPTP infringement

Meansigma methods ± standard error of mean; N=6-8 ⁺⁺=p＜0.01 is compared with control mice ^*=p＜0.01 is compared with the mice of MPTP infringement.Use one way analysis of variance and Tukey afterwards multiple comparisons check carrying out statistical analysis afterwards.

The Orally administered smilagenin of mice to the MPTP infringement significantly increased striatum GDNF and BDNF level in 60 days and significantly prevented the bonded loss of the inductive DAT of MPTP.

This embodiment has replenished the experiment in vitro among the embodiment 6 and 7 of PCT patent application WO-A-03/082893 number of incorporating this paper by reference into.Those experiment proofs with smilagenin and chinaroot greenbrier sapogenin to external remarkable prevention of the pretreatment of rat midbrain dopaminergic neuron or reverse MPP ⁺Inductive neurodegeneration.

In similar experiment; 10 weeks, big male C57 mices were accepted pump pickle or MPTP (25mg/kg/ days every day; Lumbar injection) continuous 5 days (1-5 days) and Orally administered smilagenin (10mg/kg/ days) or vehicle (the HPMC 0.5%w/v that contains tween 80 0.2%v/v) 61 days (12-72 days) or 71 days (2-72 days); Afterwards their brain is shifted out the striatum level that is used for DAT quantitatively, DAT is the sign to the degree of the neuron infringement of dopaminergic neuron.

The result is shown in hereinafter in the table 14.

Table 14 smilagenin reverses the inductive minimizing of MPTP of striatum DAT level in the mice

Meansigma methods ± standard error of mean; N=8-12, ^{* * *}=p＜0.001 is compared with control mice, ^{++ ++}=p＜0.001 is compared with the MPTP mice.Use one way analysis of variance and Fisher afterwards multiple comparisons check carrying out statistical analysis afterwards.

Compare with only accepting vectorial control mice, do not change striatum DAT level in 71 days to the Orally administered smilagenin of control mice.Orally administered smilagenin 61 of mice or the significantly inductive minimizing of MPTP of reverse striatum DAT level in 71 days to the MPTP-infringement.

Embodiment 8

Chinaroot greenbrier sapogenin and smilagenin increase the neurite in cortex, motion of the vertebra and the cognition neural unit Take place

Cortical neuron

Use the method culture of rat cortical neuron of revising of describing before (Singer waits the people, Neuroscience Letters, 1996,212,13-16 page or leaf).With chinaroot greenbrier sapogenin, smilagenin, vehicle (DMSO, 0.25%), GDNF, BDNF or NGF cultured cell 24h.For each group, in each visual field, select at random to show neuronic 15 photos of showing neurite and measure for the longest neurite of each neuron.Measure the neurite number through total neuron number in neuron number, the neuron number of not showing neurite and each visual field of counting displaying neurite.Detect six visuals field, every hole.

The result is shown in hereinafter in the table 15, is expressed as the percentage ratio that neuron number that each visual field has neurite accounts for each neuronic total number in the visual field.

The neurite that table 15 chinaroot greenbrier sapogenin and smilagenin increase in the cortical neuron takes place

Meansigma methods ± standard error of mean; Once cultivate, n=3 uses one way analysis of variance and Fisher afterwards to check afterwards and carries out statistical analysis. ^*=p＜0.01; ^{* *}=p＜0.005, ^{* * *}=p＜0.001 is compared with contrast.

Chinaroot greenbrier sapogenin and smilagenin significantly increase the length and the neuronic percentage ratio of showing neurite of existing neurite in the primary cortical neuron of rat.Be exposed to increase neurite outgrowth after chinaroot greenbrier sapogenin and the smilagenin influence can with compare with positive control GDNF, BDNF and NGF are observed.

This embodiment has replenished the experiment among the embodiment 5 of PCT patent application WO-A-03/082893 number of incorporating this paper by reference into.That experiment proof is handled the length and the neuronic percentage ratio of showing neurite that the primary cortical neuron of rat significantly increases existing neurite with chinaroot greenbrier sapogenin or smilagenin.

For whether neurotrophy, neuroprotective and the neural repairing activity of testing chinaroot greenbrier sapogenin and smilagenin depends on the for example existence of BDNF or GDNF of neurotrophic factor, carried out following experiment:

According to above-described method culture of rat cortical neuron.

Different with research before, do not add hyclone (FBS) or tire calf serum (FCS) in the culture medium, show in the culture medium not have neurotrophic factor.Add test compounds 24h.

Under the situation that does not have FBS or FCS, the rat cortical neuron is exposed to chinaroot greenbrier sapogenin, smilagenin (30nM) or vehicle (DMSO, 0.25%) one day.Use the anti-'beta '-tubulin monoclonal antibody of dilution and the anti-mouse immuning ball protein G dyeing cortical neuron of dilution.These antibody staining neuron cell bodies (quantitatively neuroprotective) and neurite (quantitatively neurotrophic effect).The every hole of epifluorescence microscope (amplification x20) with having photographing unit obtains 2 photos (10 photos of every kind of situation).Use the analysis of sum of number and cell of the cell of anti-'beta '-tubulin antibody labeling of LUCIA 6.0 softwares.

The result is shown in hereinafter in the table 16.

Table 16 chinaroot greenbrier sapogenin and smilagenin are to the neuron survival rate of the cortical neuron under serum and any other non-existent situation of neurotrophic factor, cultivated and the influence of neurite outgrowth

Meansigma methods ± standard error of mean; Use each n=12 of cultivation hole, n=2 cultivation.Use one way analysis of variance and Fisher afterwards to check afterwards and carry out statistical analysis, ^{* * *}=p＜0.001 is compared with contrast.

Chinaroot greenbrier sapogenin and smilagenin do not need neurotrophy, neuroprotective and the neural repairing activity of other neurotrophic factor to bring into play them.

The spinal motion neuron

Xaliproden (1-[2-(naphthalene-2-yl) ethyl]-4-(3-trifluoromethyl)-1; 2,5,6-tetrahydropyridine hydrochlorate); Being also referred to as SR 57746A, is chemical compound Orally active, synthetic non-peptide that is used to treat nerve retrograde affection by the Sanofi-Aventis exploitation.Xaliproden penetrates blood brain barrier and has external neurotrophic activity, and wherein it makes NGF can influence neurite outgrowth (people such as Fournier, Neuroscience, 1993,55, the 629-641 page or leaf in the PC12 cell; People such as Pradines, Journal of Neurochemistry, 1995; 64, the 1954-1964 pages or leaves) and increase mice spinal motion neuronal survival (people such as Duong, British Journal of Pharmacology; 1999,128, the 1385-1392 page or leaf).And xaliproden increases athletic performance people such as (, British Journal of Pharmacology, 1998,124, the 811-817 page or leaf) Duong of mean survival time and carrying out property motor neuron mice.The model of action of xaliproden is understood very few.As if yet the neuroprotective of xaliproden does not rely on it to serotonine _1AThe agonism of receptor (people such as Labie, British Journal of Pharmacology, 1999,127, the 139-144 page or leaf).

The nerve of following experiment contrast chinaroot greenbrier sapogenin or the anti-xaliproden of smilagenin takes place and neurite is had an effect.

According to before the method described prepare rat spine motor neuron people such as (, Neuron, 8,737-744,1992) Martinou.With chinaroot greenbrier sapogenin, smilagenin, vehicle (DMSO; 0.25%), xaliproden or BDNF cultivated after 3 days; The spinal motion neuron is cleaned twice in PBS, fixedly 5min and rinsing 3 times in PBS afterwards in the ice solution of ethanol (95%) and acetic acid (5%).Use anti-'beta '-tubulin monoclonal antibody and anti-mouse immuning ball protein G dyeing neuron.These antibody staining neuron cell bodies (quantitatively neuroprotective) and neurite (quantitatively neurotrophic effect).Examine with the fluorescent labeling staining cell.After hatching 1h, cell is cleaned in PBS 3 times.Observe culture with epifluorescence microscope with 20 x magnifications.Use is obtained a series of photos through the photographing unit of computer software control.Under same case, obtain all pictures.Use the analysis of sum (painted cell nuclei) of number and cell of the cell of anti-'beta '-tubulin antibody labeling of LUCIA 6.0 softwares.

The result is shown in hereinafter in the table 17.

The nerve that table 17 chinaroot greenbrier sapogenin and smilagenin increase in the spinal nerves motion takes place and the neurite generation

Meansigma methods ± standard error of mean; Use each n=12 of cultivation hole, n=2 cultivation.Use one way analysis of variance and Fisher afterwards to check afterwards and carry out statistical analysis, ^*=p＜0.05, ^*=p＜0.01, ^{* *}=p＜0.005 draw ^{* * *}=p＜0.001 is compared with contrast.

Data show is compared with contrast and is exposed to xaliproden and significantly increases neuron survival rate and neurite outgrowth.Chinaroot greenbrier sapogenin and smilagenin also significantly increase neuron survival rate and neurite outgrowth in the rat primitive backbone motor neuron.Increase effect that neurite takes place and be with observed those are commeasurable with positive control BDNF.

As if chinaroot greenbrier sapogenin and smilagenin promote neurogenetic effect more remarkable a little than the effect of xaliproden; Although as if compare acting in this research of chinaroot greenbrier sapogenin and smilagenin has reduced with research before.

Used amyotrophic lateral sclerosis (ALS) patient in two III clinical trial phases, to assess effectiveness and the safety (people such as Meininger of xaliproden (1 with 2mg/ days); Amyotrophic Lateral Sclerosis and Other Motor Neuron Disorders; 2004; 5, the 107-117 pages or leaves).In addition, the potentiality that xaliproden is used for Alzheimer (for this indication, do not re-use xaliproden at present and carry out (progress)) have been assessed in testing in the III phase recently.Dose-dependent side effect is main relevant with the exciting character of serotonine (5-HT) of xaliproden.

In the present embodiment, chinaroot greenbrier sapogenin and Smilagenin show to compare with xaliproden and improve or similar active overview.Important ground, chinaroot greenbrier sapogenin and Smilagenin are not 5-HT agonist and the corresponding side effect that does not show xaliproden.

This embodiment has replenished the experiment in vitro of the embodiment 8 of PCT patent application WO-A-03/082893 number of incorporating this paper by reference into.This experiment proof significantly reduces or reverses the inductive neurodegeneration of glutamate, Glu of external rat primitive backbone motor neuron with smilagenin or chinaroot greenbrier sapogenin.

Cognition neural does not have

Obtain rat cognition neural unit from conceived the 15th day Wistar rat embryo.In 37 ℃ at 5%CO ₂Cultured cell in the atmosphere of/95% air.After cultivating 2 days with chinaroot greenbrier sapogenin, smilagenin, vehicle (DMSO, 0.25%) or NGF, with cognition neural unit with twice of PBS rinsing and in 4 ℃ of paraformaldehydes (4%) that are being dissolved in PBS fixing 30min.Change cell and non-specific site is saturated thoroughly with Triton X-100 (0.1%) with hyclone.Before dyeing, with cell be dissolved in the anti-neurofilament 68 of primary antibody of the PBS that contains hyclone 5% and 200 mixture one arises from incubated at room 2h.Before the cleaning, unload microscope slide and clean twice 5min of cell with PBS, be positioned over 1h in the darkroom and be dissolved in the PBS that contains hyclone (5%) with cyanine 3 (Cy3; 1/1600) link coupled secondary antibody: anti-mouse antibodies and with fluoro isothiocyanate (FITC; 1/200) link coupled anti-rabbit antibody incubation.With PBS clean twice 5min of microscope slide and on coverslip with being dissolved in glycerol (22%) with Tris/HCl (0.2mM; PH 8.5) buffered non-oxidizability solution (9%w/v) Mowiol embedding.With microscope slide in the room temperature standing over night with hardening and be stored in the environment of lucifuge.Use has the triple optical filter microscopes of DAPI/FITC/Cy3 of x20 eyepiece and watches microscope slide.Use the every at random hole of digital camera to obtain a series of photos.

The result is shown in hereinafter in the table 18.

Table 18 chinaroot greenbrier sapogenin and smilagenin increase the neuron survival rate in the cognition neural unit

Meansigma methods ± standard error of mean; Use the n=40-49 hole, n=2 cultivation.Use one way analysis of variance and Fisher afterwards to check afterwards and carry out statistical analysis, ^*=p＜0.05 draw ^{* * *}=p＜0.001 is compared with contrast.

Chinaroot greenbrier sapogenin and smilagenin significantly increase the neuron survival rate in the primary cognition neural of the rat unit.

Embodiment 9

Chinaroot greenbrier sapogenin and the smilagenin activation endocellular transduction path identical with neurotrophic factor

The inductive neurite of chinaroot greenbrier sapogenin and smilagenin is suppressed by trk inhibitor K252a, and the neurotrophic effect of expression chinaroot greenbrier sapogenin and smilagenin is by the directly or indirectly mediation of trk receptor.This inhibition experiment is described in hereinafter and the result is shown in hereinafter in the table 19.

Like the cultivation cortical neuron of above describing in detail.Neuron is exposed to vehicle (DMSO, 0.25%) or K252a (100nM) 1h.Behind the 1h, under the situation that continues to exist at K252a, vehicle, chinaroot greenbrier sapogenin, smilagenin (30nM) or BDNF (1.85nM) are added to culture medium.Be exposed to after chinaroot greenbrier sapogenin or smilagenin (30nM), vehicle (DMSO, 0.25%) or BDNF (1.85nM) 24h, neuron cleaned with the saline (PBS) of phosphoric acid buffer and fixing in being dissolved in the glutaraldehyde of PBS (2.5%).(object lens x20 Nikon) obtains 40-60 neuronic photo showing neurite to use the photographing unit that is fixed on the microscope.Analysis to measure neurite lengths through photo.

The result is shown in hereinafter in the table 19.

The inhibition of the neurite outgrowth of the primary cortical neuron of the inductive rat of table 19 chinaroot greenbrier sapogenin and smilagenin

Meansigma methods ± standard error of mean; Use each n=81-105 neuron of cultivating, n=2 cultivation used one way analysis of variance and Fisher afterwards to check afterwards and carried out statistical analysis, ^{++ ++}=p＜0.001 is compared with contrast; ^{* * *}=p＜0.001 is compared with the identical situation that does not have K252a.

Obtain similar result in the independently experiment of use K252a, anti-BDNF antibody or the anti-GDNF antibody in cortex and midbrain neuron.

After the trk receptor activation, cause the signal specific pathway of neuronal survival to be activated, and shown that MEK1/2 participates in this path (Finkbeiner, Neuron, 2000,25,11-14 page or leaf).The inductive neurite outgrowth of smilagenin partly receives MEK1/2 inhibitor PD98059 and suppresses, and the neurotrophic effect that shows smilagenin is partly through the MEK1/2 mediation.This inhibition experiment is described in hereinafter and the result is shown in hereinafter in the table 20.

Like the cultivation cortical neuron of above describing in detail.Neuron is exposed to vehicle (DMSO, 0.25%) or PD98059 (10 μ M) 1h.After the 1h, vehicle, smilagenin (30nM) or BDNF (1.85nM) are added to culture medium under the lasting situation about existing of PD98059.Be exposed to after smilagenin (30nM), vehicle (DMSO, 0.25%) or BDNF (1.85nM) 24h, neuron cleaned with PBS and fixing in being dissolved in the glutaraldehyde of PBS (2.5%).(object lens x20 Nikon) obtains 40-60 neuronic photo showing neurite to use the photographing unit that is fixed on the microscope.Analysis to measure neurite lengths through photo.

The result is shown in hereinafter in the table 20.

The inhibition of the neurite outgrowth of the primary cortical neuron of the inductive rat of table 20 smilagenin

Meansigma methods ± standard error of mean; Use each n=86-109 neuron of cultivating, n=2 cultivation used one way analysis of variance and Fisher afterwards to check afterwards and carried out statistical analysis. ^{++ ++}=p＜0.001 is compared with contrast; ^{* * *}=p＜0.001 is compared with the identical situation that does not have PD98059.

Use the chinaroot greenbrier sapogenin that produces analog result to carry out similar experiment.

CAMP response element conjugated protein (CREB) belongs to transcription factor family and very important in regulating neuronal survival.In addition, after the trk receptor activation, CREB is raised (Finkbeiner, Neuron, 2000,25,11-14 page or leaf).Chinaroot greenbrier sapogenin significantly increases the amount of the CREB (pCREB, the activity form of CREB) of phosphorylation in Chinese hamster ovary (CHO) cell.This experiment is described in hereinafter and the result is shown in hereinafter in the table 21.

CHO is hatched 24h with DMSO (0.5%) or chinaroot greenbrier sapogenin (10 μ M).Afterwards cell is cleaned with cold PBS, in lauryl sulfate (SDS) buffer, dissolve, boil 5min and measure protein content through the Bradford method.Afterwards sample is separated on sds page and be transferred to PVDF (Bio-Rad) film.In 5% defatted milk powder, expose after the 1h, with film in 4 ℃ in primary antibody mice pCREB (Upsdate, 1: 1000) and the middle incubated overnight of mice beta-actin (Santa Cruz, 1: 1000).The secondary antibody that film is closed at the peroxidase yoke in room temperature (Wuhan Boster Biology Technology, China.1: hatched 2000) 1 hour and develop the color afterwards with ECL reagent (Pierce).Divide band with film through in 2 mercapto ethanol (100mM), SDS (2%), Tris HCl (62.5mM), hatching 30min in pH 6.8 and 50 ℃.(Gel Doc 2000, it is quantitative Bio-Rad) to carry out the density of immunostaining with the picture analyzing appearance to use Image J analytical system.With the relative quantity of the immunostaining of the pCREB of every band to the beta-actin band standardization that in identical experiment, moves and be expressed as arbitrary unit.

Table 21 is exposed to the expression of the CREB of phosphorylation behind the chinaroot greenbrier sapogenin 24h in Chinese hamster ovary celI

Meansigma methods ± standard error of mean; N=5, through paired t check carrying out statistical analysis, ⁺⁺=p＜0.001 is compared with contrast.

Embodiment 10

Pretreatment with chinaroot greenbrier sapogenin, smilagenin, table chinaroot greenbrier sapogenin and table smilagenin Reduce the inductive infringement of glutamate, Glu to cortical neuron

The primary cortical neuron of rat is exposed to glutamate, Glu increases lactic acid dehydrogenase (LDH) activity that glutamate, Glu exposes 24h measurement afterwards, shows significant neuron infringement.Use the method culture of rat cortical neuron of revising of describing before (Singer waits the people, Neuroscience Letters, 1996,212,13-16 page or leaf).At the 10th day that cultivates, culture medium is changed to the defined medium of serum-free.At the 12nd day, culture is cleaned and is positioned over 24h in the fresh culture that contains test compounds or vehicle (DMSO, 0.25%).At the 13rd day, neuron is exposed to glutamate, Glu (100 μ M in 37 ℃; 10min).Before measuring LDH, culture is supplemented with other 24h in test compounds or the vectorial fresh culture with being supplemented with the cleaning of test compounds or vectorial fresh culture and being positioned over afterwards.LDH activity through 24h after glutamate, Glu exposes measures in the culture medium is assessed the neuron infringement.

The result is shown in hereinafter table 22 in 26.

Table 22 chinaroot greenbrier sapogenin reduces the inductive infringement of glutamate, Glu in the cortical neuron

Cortical neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±2.23
		Glutamate, Glu (100 μ M)	65.83±2.46 +++
Glutamate, Glu+chinaroot greenbrier sapogenin (1nM)	76.88±2.79***
		Glutamate, Glu+chinaroot greenbrier sapogenin (3nM)	77.23±2.62***
Glutamate, Glu+chinaroot greenbrier sapogenin (10nM)	73.50±3.05*
		Glutamate, Glu+chinaroot greenbrier sapogenin (30nM)	78.91±2.97***
Glutamate, Glu+chinaroot greenbrier sapogenin (100nM)	76.30±4.15***

Meansigma methods ± standard error of mean; Use each n=4 of cultivation hole, 3 cultivations are used one way analysis of variance and Fisher afterwards to check afterwards and are carried out statistical analysis. ⁺⁺⁺=p＜0.005 is compared with contrast ^*=p＜0.05; ^*=p＜0.01; ^{* *}=p＜0.005; Compare with glutamate, Glu.

Table 23 smilagenin reduces the inductive infringement of glutamate, Glu in the cortical neuron

Cortical neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±4.17
		Glutamate, Glu	67.09±3.46 +++
Glutamate, Glu+smilagenin (1nM)	81.53±1.66***
		Glutamate, Glu+smilagenin (3nM)	78.19±1.85**
Glutamate, Glu+smilagenin (10nM)	82.50±1.00***
		Glutamate, Glu+smilagenin (30nM)	89.86±3.55***
Glutamate, Glu+smilagenin (100nM)	82.45±2.18***

Meansigma methods ± standard error of mean; Use the n=4 hole, 1 cultivation.

Table 24 table chinaroot greenbrier sapogenin reduces the inductive infringement of glutamate, Glu in the cortical neuron

Cortical neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±4.17
		Glutamate, Glu	67.09±3.46 +++
Glutamate, Glu+table chinaroot greenbrier sapogenin (1nM)	84.79±2.40***
		Glutamate, Glu+table chinaroot greenbrier sapogenin (3nM)	80.39±5.18*
Glutamate, Glu+table chinaroot greenbrier sapogenin (10nM)	83.80±4.18***
		Glutamate, Glu+table chinaroot greenbrier sapogenin (30nM)	87.17±2.51***
Glutamate, Glu+table chinaroot greenbrier sapogenin (100nM)	86.42±2.95***

Meansigma methods ± standard error of mean; Use the n=4 hole, 1 cultivation.

Table 25 table smilagenin reduces the inductive infringement of glutamate, Glu in the cortical neuron

Cortical neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±5.18
		Glutamate, Glu	70.15±1.07 +++
Glutamate, Glu+table smilagenin (3nM)	82.49±3.93**
		Glutamate, Glu+table smilagenin (10nM)	78.57±2.15
Glutamate, Glu+table smilagenin (30nM)	81.76±2.09**
		Glutamate, Glu+table smilagenin (100nM)	78.39±1.75
Glutamate, Glu+table smilagenin (300nM)	78.86±1.80*

Meansigma methods ± standard error of mean; Use the n=4 hole, 1 cultivation.

Table 26 diosgenin does not reduce the inductive infringement of glutamate, Glu in the cortical neuron

Cortical neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±4.20
		Glutamate, Glu	67.67±4.54 +++
Glutamate, Glu+diosgenin (3nM)	69.43±1.76
		Glutamate, Glu+diosgenin (10nM)	66.51±5.13
Glutamate, Glu+diosgenin (30nM)	68.98±5.39
		Glutamate, Glu+diosgenin (100nM)	70.95±5.03
Glutamate, Glu+diosgenin (300nM)	75.02±2.68

Meansigma methods ± standard error of mean; Use the n=4 hole, 1 cultivation.

In the primary cortical neuron of rat; Compare with the neuron that only is exposed to glutamate, Glu, 24h used chinaroot greenbrier sapogenin, smilagenin, table chinaroot greenbrier sapogenin (1-100nM) and the inductive LDH of table smilagenin (3-300nM) pretreatment significantly minimizing glutamate, Glu to discharge before glutamate, Glu exposed.

On the contrary, 24h used diosgenin (3-300nM) pretreatment not prevent the neuron infringement before glutamate, Glu exposed.

The activity of chinaroot greenbrier sapogenin, smilagenin, table chinaroot greenbrier sapogenin and table smilagenin reaches plateau at nanomolar concentration but does not cause any toxicity.The test compounds of micro-molar concentration is settled out from solution under these experiment conditions.

This embodiment has replenished the experiment in vitro of the embodiment 2 to 4 of PCT patent application WO-A-03/082893 number of incorporating this paper by reference into; Those experiment proofs are used the remarkable prevention of pretreatment of chinaroot greenbrier sapogenin, table chinaroot greenbrier sapogenin, smilagenin, table smilagenin or its 3-ketone or 3-ester or are reversed the inductive neurodegeneration of glutamate, Glu, yet diosgenin does not show such activity.

Embodiment 11

Chinaroot greenbrier sapogenin, table chinaroot greenbrier sapogenin, smilagenin and the different Rhizoma Smilacis Chinensis of table in the dopaminergic neuron The anti-apoptotic effect of ruscogenin

Like the culture of rat dopaminergic neuron above described in detail people such as (, Journal of Neurochemistry, 1988,50, the 1900-1907 page or leaf) Schinelli.At the 5th day; With culture with being supplemented with test compounds (30nM), vehicle (DMSO; 0.25%) or the fresh culture of the combination of BDNF (1.85nM) and GDNF (0.17nM) clean and be positioned over other 24h in the fresh culture of the combination that is supplemented with test compounds (30nM), vehicle (DMSO, 0.25%) or BDNF (1.85nM) and GDNF (0.17nM).

Compare with contrast, the primary dopaminergic neuron of rat is exposed to MPP ⁺(2 μ M 24h) cause reduction on the dopaminergic neuron number.At the 6th day, under the situation that the combination of test compounds, vehicle or BDNF and GDNF exists with MPP ⁺(2 μ M) added to culture other 48 hours.MPP ⁺Induce neuronal death through inhibition complex I and ATP subsequently disappearance in mitochondrion, cause free atomic group to produce and apoptosis induced.After incubation period, that culture is fixing with the paraformaldehyde (4%) that is dissolved in PBS.After fixing, neuron is changed 30min thoroughly with Triton x 100 (0.05%).Afterwards neuron and anti-tyrosine hydroxylase (TH) antibody one are arised from 37 ℃ and hatch 2h.Neuron is cleaned three times with PBS and use goat anti-mouse antibody/Cy3 to hatch 2h afterwards in 37 ℃.With the neuron embedding and use fluorescence microscopy.

The result is shown in hereinafter in the table 27.

Table 27 chinaroot greenbrier sapogenin, smilagenin, table chinaroot greenbrier sapogenin and table smilagenin reduce MPP ⁺Induce the loss of midbrain dopaminergic neuron

Dopaminergic neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±3.20
		+MPP +(2μM)	55.31±3.15 ++++
+MPP ++ chinaroot greenbrier sapogenin (30nM)	88.73±4.39****
		+MPP ++ smilagenin (30nM)	97.91±3.63****
+MPP ++ table chinaroot greenbrier sapogenin (30nM)	95.01±4.52****
		+MPP ++ table smilagenin (30nM)	115.12±4.73****
+MPP ++BDNF(1.85nM)&GDNF(0.17nM)	121.94±6.51****

Meansigma methods ± standard error of mean; Use each n=40 of cultivation or 80 visuals field, hole, n=1 or cultivate for 2 times.Use one way analysis of variance and Dunnett afterwards to check afterwards and carry out statistical analysis; ^{++ ++}=p＜0.005 is compared with contrast, ^{* * *}=p＜0.005; With MPP ⁺Compare.

Chinaroot greenbrier sapogenin, smilagenin, table chinaroot greenbrier sapogenin and table smilagenin significantly prevent MPP ⁺The neuronic minimizing of Induction of dopaminergic.The combination that is exposed to neurotrophic factor BDNF and GDNF also significantly prevents MPP ⁺The neuronic minimizing of Induction of dopaminergic.

Embodiment 12

Glutamate, Glu or MPP ⁺ The nerve reparation of chinaroot greenbrier sapogenin and smilagenin after the inductive infringement The property

Cortical neuron

The important goal of the processing of neurodegeneration disease is not only to prevent to develop to reverse the neurone loss that takes place among the patient in addition.The primary cortical neuron of rat is exposed to glutamate, Glu (100 μ M; 10min) afterwards, chinaroot greenbrier sapogenin and smilagenin (30nM) 24h after handling significantly reverses the inductive infringement of glutamate, Glu.This embodiment has developed the work of report among the embodiment 2 to 4 of PCT patent application WO-A-03/082893 number.

Like the preparation rat cortical neuron of above introducing in detail.At the 13rd day, with culture in confirming the culture medium of composition in 37 ℃ at 5%CO ₂Be exposed to glutamate, Glu (100 μ M) 10min in/95% air atmosphere.After incubation period, culture is cleaned in containing chinaroot greenbrier sapogenin, smilagenin or vectorial fresh culture and keep.Cell, is cultivated glutamate, Glu other 24h and the assessment neuron infringement as above describing in detail afterwards after exposing.

The result is shown in hereinafter in the table 28.

Table 28 chinaroot greenbrier sapogenin and smilagenin reverse the inductive infringement of glutamate, Glu in the cortical neuron

Cortical neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±4.03
		+ glutamate, Glu (100 μ M)	66.32±2.36 ++++
+ glutamate, Glu+chinaroot greenbrier sapogenin (30nM)	103.43±5.10****
		+ glutamate, Glu+smilagenin (30nM)	111.06±3.40****

Meansigma methods ± standard error of mean; Use each n=4 of cultivation hole, 2 cultivations are used one way analysis of variance and Fisher afterwards to check afterwards and are carried out statistical analysis; ^{++ ++}=p＜0.001 is compared with contrast, ^{* * *}=p＜O.001 compare with glutamate, Glu.

The spinal motion neuron

Like the preparation rat spine motor neuron of above introducing in detail.At the 10th day, culture medium removed and with culture in the culture medium of confirming composition in 37 ℃ at 5%CO ₂Be exposed to glutamate, Glu (4 μ M) 10min in/95% air atmosphere.After glutamate, Glu exposes, culture is positioned in the fresh culture that contains chinaroot greenbrier sapogenin, smilagenin, vehicle or BDNF after 37 ℃ are cleaned with DMEM.After the 48h, like the degree of the definite motor neuron infringement above described in detail.This embodiment has developed the work of report among the embodiment 8 of PCT patent application WO-A-03/082893 number.

The result is shown in hereinafter in the table 29.

Table 29 chinaroot greenbrier sapogenin and smilagenin reverse the inductive infringement of glutamate, Glu in the spinal motion neuron

The spinal nerves motion
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±8.87
		+ glutamate, Glu (4 μ M)	75.52±2.58+
+ glutamate, Glu+chinaroot greenbrier sapogenin (0.03nM)	101.09±4.12****
		+ glutamate, Glu+chinaroot greenbrier sapogenin (3nM)	108.10±3.56****
+ glutamate, Glu+chinaroot greenbrier sapogenin (300nM)	120.43±7.46****
		+ glutamate, Glu+smilagenin (0.03nM)	90.98±2.46*
+ glutamate, Glu+smilagenin (3nM)	101.53±3.18****
		+ glutamate, Glu+smilagenin (300nM)	106.61±4.24****
+ glutamate, Glu+BDNF (3nM)	106.60±6.14****

Meansigma methods ± standard error of mean; Use each n=6 of cultivation hole, cultivate and be used for n=1 time of BDNF for 2 times and cultivate, use one way analysis of variance and Fisher afterwards check afterwards and carry out statistical analysis. ⁺=p＜0.005 is compared with contrast, ^{* * *}=p＜0.001, ^*=p＜0.05 is compared with glutamate, Glu.

Rat primitive backbone motor neuron is exposed to glutamate, Glu (4 μ M; 10min) increase the LDH activity that glutamate, Glu exposes 48h measurement afterwards, show significant neuron infringement.Chinaroot greenbrier sapogenin and smilagenin (0.03-300nM) 48h after processing significantly reverses the inductive infringement of glutamate, Glu.

Yet the reverse of the infringement that is provided by the chinaroot greenbrier sapogenin and the smilagenin of least concentration is different between culture, and expression 0.03nM can be an active lower limit in this model.Brain derived neurotrophic factor (3nM) is compared the inductive LDH of remarkable reverse glutamate, Glu as positive control and with the spinal motion neuron that only is exposed to glutamate, Glu active.

Dopaminergic neuron

The primary dopaminergic neuron of preparation rat described above.At the 5th day, in culture medium in 37 ℃ at 5%CO ₂In/95% air atmosphere with MPP ⁺(2 μ M) adds to culture 24h.Compare with contrast, the primary dopaminergic neuron of rat is exposed to MPP ⁺(2 μ M 24h) cause the remarkable reduction of dopaminergic neuron number.The 6th day, culture medium is removed and is added the fresh culture of the combination that contains vehicle (DMSO, 0.25%), chinaroot greenbrier sapogenin, smilagenin or BDNF and GDNF.After the 48h, like the degree of the definite dopaminergic infringement above described in detail.

The result is shown in hereinafter in the table 30.

Table 30 chinaroot greenbrier sapogenin and smilagenin reverse MPP in the dopaminergic neuron ⁺Inductive infringement

Dopaminergic neuron
		Condition	Neuron survival rate (% of contrast)
Contrast	100.00±5.79
		+MPP +(2μM)	75.81±4.00 +++
+MPP ++ chinaroot greenbrier sapogenin (30nM)	104.30±5.63****
		+MPP ++ smilagenin (30nM)	113.44±4.62****
+MPP ++BDNF(1.85nM)&GDNF(0.17nM)	97.85±4.68***

Meansigma methods ± standard error of mean; Use n=40 the visual field, n=1 cultivation used one way analysis of variance and Fisher afterwards to check afterwards and carried out statistical analysis. ⁺⁺⁺=p＜0.005 is compared with contrast, ^{* *}=p＜0.005, ^{* * *}=p＜0.001 and MPP ⁺Compare.

Data show is exposed to chinaroot greenbrier sapogenin and smilagenin (30nM) significantly reverses MPP in the dopaminergic neuron ⁺Inductive decline.Be exposed to the same remarkable MPP in the dopaminergic neuron that reverses of combination of neurotrophic factor BDNF (1.85nM) and GDNF (0.17nM) ⁺Inductive decline.

In similar experiment, its result is shown among Fig. 1, and the variable concentrations of inspection smilagenin reverses MPP in the primary dopaminergic neuron of rat ⁺(2 μ M, 24h) effect of inductive neuron infringement.BDNF, GDNF and vectorial concentration are as stated.The dopaminergic culture is being contained smilagenin (0.3fM to 30nM), hatching 24 hours in the culture medium of the combination of BDNF (1.85nM) and GDNF (0.17nM) or vehicle (DMSO, 0.25%).Afterwards with MPP ⁺(2 μ M) or vehicle add to culture medium and culture are hatched other 48h.Dopaminergic in each visual field (TH positive) neuron number can be combined to himself reference standardization so that data quantitatively and afterwards through immunohistochemistry and fluorescence microscope.Be exposed to MPP ⁺Use smilagenin (3fM-30nM) to handle 48h after the 24h and significantly reverse MPP ⁺Inductive neuron damages, and has the EC of 13.4fM ₅₀

Embodiment 13

Refreshing in the mouse model of the Orally administered improvement nerve damage evil of chinaroot greenbrier sapogenin and smilagenin Recovery (pmn mice) through function

Carrying out property motor neuron (pmn) mice is the genetic model of degenerative motor neuron disease; Comprise atrophy (dying-back) process of following the far-end axonai degeneration and the relative reservation (people such as Schmalbruch of near-end aixs cylinder and cyton; Journal of Neuropathology and Experimental Neurology; 1991; 50, the 192-204 pages or leaves).The homozygous cranium tail (caudio-cranial) of suffering from the motion aixs cylinder of pmn/pmn is degenerated and a few week death after birth; Maybe be because the respiratory muscle denervate (people such as Schmalbruch; Journal of Neuropathology and Experimental Neurology; 1991; 50, the 192-204 pages or leaves; People such as Sendtner, Nature, 1992,358,502-504 page or leaf).Although the pmn mice can not be considered to the accurate animal model (people such as Kennel of any duplicate of any special human motion neuronal disease; Neurobiology of Disease; 1996; 3; But its representative assessment is used for the useful model of the new drug candidates potentiality of nerve retrograde affection the 137-147 page or leaf).This mouse model has been used to confirm (the people such as Sagot of potential mechanism of causing a disease under the motor neuron degeneration; Journal of Neuroscience; 1995; 15; The 7727-7733 page or leaf) and improve and to be used to handle the for example potential therapeutic strategy of ALS, progressive myatrophy, Duchenne-Arandisease, PBP, pseudobulbar palsy and primary lateral sclerosis (people such as Haase, Nature Medicine, 1997 of motor neuron disease; 3, the 429-436 pages or leaves; People such as Sendtner, Nature, 1992,358,502-504 page or leaf; People such as Sagot, Journal of Neuroscience, 1995,15,7727-7733 page or leaf; People such as Sagot, Journal of Neuroscience, 1996,16,2335-2341 page or leaf).This embodiment has developed the work of report among the embodiment 11 of PCT patent application WO-A-03/082893 number of incorporating this paper by reference into.

Affected homozygous+/+pmn (" pmn mice ") mice from Neurofit (Illkirch, the extra toe locus (Xt) of France) breeding+/+breeding colonies of the two heterozygous mices of pmn obtains.From being born beginning in back 10 days just after the initial symptom of disease manifests, raise by force the administration of pmn mice through the oral of every day.Chinaroot greenbrier sapogenin (0.03,0.3 and 3 μ g/kg/ days) is applied to the pmn mice as the suspension in oil (10ml/kg).Use standard N euromatic 2000M electromyograph(EMG instrument to carry out electromyogram (EMG) record according to the guide of U.S. electrodiagnosis association (American Association of Electrodiagnostic Medicine).Carried out the athletic performance of criterion behavior test (grid, rotation and suspension test) assessment pmn mice weekly since the 8th day.

Chinaroot greenbrier sapogenin is assessed (CMAP, the indirect measurement of functional movement neuron number) to the amplitude of the motion response that the influence of motor function is brought out through the record gastrocnemius

The result is shown among Fig. 2 of accompanying drawing.

The pmn matched group shows the rapid reduction of 12 days amplitudes of CMAP when big.Postpone the degeneration (p＜0.001) of neuronal function to pmn mice Orally administered chinaroot greenbrier sapogenin every day (0.3 μ g/kg/ days).The number of times of falling down of control mice performance increased sharply when big from 12 days.Compare with the pmn matched group, significantly postpone the degeneration (p=0.02 MANOVA) on rotation and the grid behavioral test to pmn mice Orally administered chinaroot greenbrier sapogenin every day (0.3 μ g/kg/ days).Compare with the pmn matched group, Orally administered chinaroot greenbrier sapogenin every day (0.3 μ g/kg/ days) significantly increases the survival rate of pmn mice and (compares up to 62% logarithm order, χ with contrast ²=7.36, p=0.006).

By contrast, do not postpone the carrying out of motor neuron degeneration in this genetic model with minimum proof load Orally administered chinaroot greenbrier sapogenin (0.3 μ g/kg/ days) every day after the clinical symptoms outbreak.

These results show, and induced by neurotrophin thing chinaroot greenbrier sapogenin Orally active, non-peptide can postpone the carrying out of motor neuron degeneration in this genetic model.In the pmn mouse model, tested neurotrophic factor (ciliary neurotrophic factor; CNTF; People such as Sagot, Journal of Neuroscience, 1995,15,7727-7733 page or leaf) and GDNF (people such as Sagot, Journal of Neuroscience, 1996,16, the 2335-2341 page or leaf).These researchs show that CNTF (lumbar injection of CNTF secretory cell is used) increases the time-to-live 40% and improves motor function;, do not slow down GDNF disease (people such as Sagot, Journal of Neuroscience, 1996 yet improving the motor neuron survival; 16, the 2335-2341 pages or leaves).Neurotrophic factor has been considered to be used for the processing of motor neuron disease; Yet their clinical widely uses are very problematic as protein.From carry out of birth, improve mouse movement performance and life-span (～50% to the neurotrophy compound S R 57746A (xaliproden) of the Orally administered non-peptide of pmn mice lag motion neurodegeneration; People such as Duong, British Journal of Pharmacology, 1998,124,811-817 page or leaf).And Orally administered CGP 3466B (anti-apoptosis agent) postpones the carrying out of disease and improves pmn mouse life 57% people such as (, British Journal of Pharmacology, 2000,131, the 721-728 page or leaf) Sagot when seizure of disease; Molecule BN 80933 (neuronal nitric oxide synthase and lipid peroxidation inhibitor) improves pmn mouse life 40% people such as (, British Journal of Pharmacology, 2000,131, the 721-728 page or leaf) Sagot

Importantly, when after the symptom in disease during Orally administered chinaroot greenbrier sapogenin, chinaroot greenbrier sapogenin postpones the carrying out of disease and improves the pmn mouse life up to 62% in the body in this model.Obtain similar result with smilagenin.

Embodiment 14

The other mice mould of the Orally administered improvement nerve damage evil of chinaroot greenbrier sapogenin and smilagenin The recovery of function of nervous system in the type (neural extrusion mode)

The sciatic nerve extrusion mode is the reversible model (McMahon and Priestley, Current Opinion in Neurobiology, 1995,5,616-624 page or leaf) that is used for the well-characterized of nerve injury after motor neuron disease and the wound.The nerve damage evil produces through the mechanical pressure that uses mosquito forceps (using twice, the sciatic trifurcation 5mm apart from the right of these mices).The local inflammation of the nerve around this causes the nerve degeneration of two time-of-weeks and continues nearly afterwards.Function of nervous system be lost in mechanical damage after the 4-5 time-of-week restore gradually.

After sciatic nerve is undermined; To C57 mice Orally administered chinaroot greenbrier sapogenin every day (3mg/kg/ days; Be dissolved in and orally in the oil raise by force) and smilagenin (0.3 with 3mg/kg/ days; Be dissolved in and orally in the oil raise by force) 6 weeks significantly improved the recovery of function of nervous system; As measured in the gastrocnemius through CMAP parameter (amplitude, incubation period and persistent period are respectively the functional indirect indicators of active motor fiber, motor nerve conduction velocity and nerve fiber) and sciatic morphological analysis (ratio of the fiber of degeneration).The 4-methyl catechol is used as positive control (people such as Kaechi, Journal of Phamacology and Experimental Therapeutics, 1995,272, the 1300-1304 page or leaf).

The result is shown among Fig. 3 of accompanying drawing.

With ketamine hydrochlorate (60mg/kg, lumbar injection) with C57bl/6 RJ mouse anesthesia.Sciatic nerve is pushed middle burst of horizontal surgical exposure and at the about 5mm of the sciatic trifurcation of distance place.Nerve was pushed twice 30 seconds with mosquito forceps, and 90 degree rotations between each extruding.The local inflammation of the nerve around this causes the nerve degeneration of two time-of-weeks and continues nearly afterwards.Function of nervous system be lost in mechanical damage after the 4-5 time-of-week restore gradually.Like described assessment electromyogram record above.

The result is shown in hereinafter in the table 31.

Table 31 chinaroot greenbrier sapogenin and smilagenin reduce the number of the fiber of degenerating in the mouse model of nerve damage evil

Group	The fiber (% of contrast) of degenerating
		Contrast	0.00±6.71
Neural extruding	10910+265 ++++
		Chinaroot greenbrier sapogenin (3mg/kg/ days)	33.50±12.60****
Smilagenin (0.3mg/kg/ days)	16.70±4.23****
		Smilagenin (3mg/kg/ days)	-8.10±6.03****
4-methyl catechol (10 μ g/kg/ days)	-7.48±1.62****

Meansigma methods ± standard error of mean; Use one way analysis of variance and Dunnett afterwards to check the statistical analysis that carries out modified fibre, n=3-4 afterwards. ^{++ ++}=p＜0.001 is compared with contrast, ^{* * *}=p＜0.001 is compared with the nerve extruding.

Chinaroot greenbrier sapogenin and smilagenin are Orally active and recovery that can improve function of nervous system and stimulate the neuranagenesis in the sciatic nerve extrusion mode.

Embodiment 15

Chinaroot greenbrier sapogenin and smilagenin in old animal, reduce anxiety and repair cognitive competence with The decline of BDNF

To the Orally administered chinaroot greenbrier sapogenin of old Sprague Dawley (SD) rat (20 months big) or smilagenin (18mg/kg/ days) 3 months.The SD rat of youth (4 months big) will old SD rat (20 months big) as healthy positive control be used as the contrast of neurodegeneration.Y type labyrinth is used to assess learning and memory, and according to the model that causes the character of the test of the distress of animal is considered too anxiety.Bottom at each arm in Y type labyrinth is the array (2mmx140mm) of the copper post that the electric current of adjustable voltage is applied to it when needing.Each arm is that 450mm is long, has the 15w bulb endways.After the bimestrial processing, every rat was trained continuous 7 days, once a day.At each training period, rat is placed in the arm in Y type labyrinth and after the 2min electric current is used for the copper post of counterclockwise arm and lights the zone that clockwise bulb is represented non-energising.If rat runs into the arm of lighting, write down once correct response, otherwise the once wrong response of record.This stimulation-response test repeats 20 times every day, has the pause of 5s between each test.The number of times of the correct response of record and total time period of 20 tests.The number of times that calculates correct response is divided by the merchant of total response time and as the index of learning capacity, it is high more to discuss high more learning capacity.Behind the learning test one month (3 months processing), carry out Y labyrinth test once more and the index of memory ability is made in the commercialization that will obtain.When processing finishes, mice killed and brain shifted out and be used for using the quantitative BDNF of ELISA (data of BDNF in above embodiment 3).

The result of Y type maze experiment is shown in hereinafter in the table 32.

Table 32 chinaroot greenbrier sapogenin and smilagenin are repaired cognitive competence (learning and memory) and the decline of BDNF level in the old rat

Meansigma methods ± standard error of mean; N=9-10 uses paired single tail student t check carrying out statistical analysis, ^{++ ++}=p＜0.001 is compared with contrast, ^{* * *}=p＜0.001 is compared with old rat.

By oral administration to old rat nearly 3 months chinaroot greenbrier sapogenin and smilagenin (18mg/kg/ days) minimizing anxiety, with cognitive competence (learning and memory ability) to observed reparation in young rats.

Embodiment 16

Orally administered chinaroot greenbrier sapogenin and smilagenin are delivered to many bodily tissues

That chinaroot greenbrier sapogenin and smilagenin have been proved to be Orally active and be to have measured after Orally administered in Rodents and the non-Rodents their blood plasma, brain, the concentration of spinal cord (with its hetero-organization)

The result is shown in hereinafter in the table 33.

Chinaroot greenbrier sapogenin and smilagenin are dispersed to blood plasma, brain and spinal cord after table 33 single oral administration

Orally administered chinaroot greenbrier sapogenin and smilagenin travel to the neuron position and the blood plasma of health.

Embodiment 17

Orally administered chinaroot greenbrier sapogenin and smilagenin are nontoxic in effective dose

Chinaroot greenbrier sapogenin and smilagenin are external to be that activated and lower concentration is the perhaps variable activity of performance in neuron of non-activity at nanomolar concentration.The toxicity that the higher concentration observation in vitro that is not presented at neurotrophic factor at the chinaroot greenbrier sapogenin and the smilagenin of higher concentration arrives.

Used chinaroot greenbrier sapogenin (in the rat up in 26 weeks and the non-Rodents up to 39 weeks) and smilagenin (in mice and the non-Rodents up to 52 weeks) to carry out secular toxicity research after high dose Orally administered, in a single day toxicity that the high level that does not have demonstration to reach neurotrophic factor will manifest or any sign of adverse events.

Embodiment 18

Parkinson in the macaque of smilagenin minimizing MPTP infringement is also regulated in the shell nuclear GDNF and BDNF concentration

Make 19 female machins (4-6 year is big for Macaca fascicularis, 3.0-4.5kg) adapt to experimental situation and assess the baseline behavior with program 3 months and in all animals.14 female macaques are accepted MPTP (0.2mg/kg/ days, subcutaneous) and significantly stable parkinson disease symptom up to developing.Animal (n=7/ group) is set to two groups at random; (HPMC, 0.5%w/v contains Tween 80,0.2%v/v) for smilagenin (20mg/kg/ days, oral) or vehicle contrast.Use vehicle and be used as matched group to 5 macaques not accepting MPTP.

MPTP uses with the smilagenin or the vehicle in 18 week are used the assessment of carrying out Parkinsonian anergy afterwards afterwards.By estimating the parkinson disease anergy through the postmortem analysis of DVD record to handling unwitting neuropathist.When research finishes, shift out and use multiple ELISA/Aushon to measure the brain to measure the level of unconjugated GDNF and BDNF in the shell nuclear.The macaque that the MPTP that uses in this experiment handles is provided for the model of generally acknowledging of parkinson and similar motion-sensory nerve degeneration patient's condition thus.

The result is shown in hereinafter in the table 34.

The improvement behavior and regulate GDNF and the shell nuclear level of BDNF in the macaque of MPTP infringement of table 34 smilagenin

###=p＜0.001 is used back parkinson disease anergy with MPTP and is compared.The two-way variance analysis of use non-matching and Bonferroni multiple comparisons are checked afterwards and are carried out statistical analysis. ^*=p＜0.01 is compared with the macaque of MPTP infringement.Use one way analysis of variance and Tukey afterwards multiple comparisons check carrying out statistical analysis afterwards.

The level of parkinson's syndrome in the macaque that significantly reduces the MPTP infringement Orally administered 18 weeks of smilagenin of the macaque of MPTP infringement.With receive vectorial macaque and compare GDNF and BDNF level in the shell nuclear of the same macaque that significantly reduces the MPTP infringement of smilagenin to not having a remarkable different level with contrast is observed in the macaque of infringement.

These data show that smilagenin is the normalization effect under chronic administration to the effect of GDNF and BDNF expression; That is, have through level being restored watch for animals and avoid secular regulating action the excessive exposure of GDNF and BDNF to approximately normal state.Remarkable reduction in this variation and the macaque on the parkinson's syndrome level is very relevant.

Discuss

Above embodiment proves that A/B cis spirostane steroid sapogenines chinaroot greenbrier sapogenin and smilagenin are the induced by neurotrophin agent, as through external and stripped data are proved; They are external and interior neuroprotective of body and neural reparation property for Orally administered back.They do not need the existence of neurotrophic factor as derivant and work, and so seemingly real NF derivant rather than NF reinforcing agent.

Using of neurotrophic factor (for example BDNF and GDNF) is the strategy that disease is regulated in depression, schizophrenia, parkinson and other diseases, and this strategy has solid scientific theory.Yet verified be difficult to scientific theory is converted to clinical.This is to draw from the protein properties of neurotrophic factor with based on inherent difficulty the genes matter delivering method of operation, viral vector and cell.The amount of effectiveness that the complicated independent factor of nutritional need possibility restricted passage of neuron obtains and required trophic factors and same at present the unknown of persistent period of the needed processing of acquisition clinical benefit.

The correlation molecule that defines among the induced by neurotrophin agent chinaroot greenbrier sapogenin of Orally active and smilagenin and the application has overcome many in these difficulties.We this shown chinaroot greenbrier sapogenin and smilagenin external be activated at picomole and nanomolar concentration.They do not have the viewed toxicity of higher concentration of display body extrinsic nerve trophic factors.

Disclosed evidence shows together before in the list of references that evidence among the application and this paper quote; With limited and manageable side effect induce NF for example the stable state of self regulating of BDNF and/or GDNF will be selected from A/B-cis furostan through what use effective amount to the curee; Furan steroid alkene; Spirostane and spirostene steroid sapogenines and its ester; Ether; Ketone and glycosylated form at least a dose and obtain, and this will be many disease and patient's condition that are used to handle and prevent NF mediation neurological for example; Psychiatry; Inflammatory; Allergia; Immunity; New and beyond thought benefit are provided in the treatment of tumprigenicity and related conditions and the method for non-treatment.

The present invention above extensively and has without limitation been described.Very obvious variation is included in by in the defined scope of the present invention of appended claim with the modification expection for a person skilled in the art.

Claims (22)

1. in the curee, induce the for example method of the stable state of self regulating of BDNF and/or GDNF of NF through under homeostatic control, regulating curee's natural neurotrophic factor (NF) for one kind with avirulent mode, said method comprise to the curee use effective dose be selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines with and one or more agent of ester, ether, ketone and glycosylated form.

2. method according to claim 1, wherein the stable state of self regulating of NF said induced to follow with the NF limited and manageable side effect that for example NGF excessive induced, excessive stimulation or excessive enhancing are relevant and the side effect relevant with receptor (antagonism) agonism and the side effect relevant with the enzyme combination and takes place.

3. method according to claim 1; The processing of wherein said method and the disease that is used for NF mediation or the method for prevention are united use its human or inhuman mammal of needs; The processing or the prevention of the disease of said NF mediation for example are selected from: (a) processing of neurological's disease or prevention; Said neurological's disease for example is selected from: dementia; The cognitive impairment that age is relevant; Alzheimer; The alzheimer disease of Alzheimer type (SDAT); Dementia with Lewy body; Vascular dementia; Parkinson; Postencephalitic parkinsonism; Parkinson with the reason except that postencephalitic and parkinson; Muscular dystrophy comprises facioscapulohumeral muscular dystrophy (FSH); Duchenne muscular dystrophy; Duchenne muscular dystrophy and Bu Lusishi muscular dystrophy; Fuchs; Myotonic dystrophy; Cerneal dystrophy; Reflex sympathetic dystrophy syndrome (RSDSA); The neural blood vessel malnutrition, myasthenia gravis, the Eton, Lambert is sick; Huntington Chorea; Motor neuron disease comprises amyotrophic lateral sclerosis (ALS), baby's spinal cord amyotrophy, multiple sclerosis; Postural hypotension; Pain, neuralgia, traumatic neurodegeneration are for example after apoplexy or after unexpected (for example traumatic head or brain injury or spinal cord injury); Crust Teng Shi is sick; Cockayne syndrome, mongolism, the degeneration of cortex ganglion basal; Multiple system atrophy; Brain atrophy, olivopontocerebellar atrophy, dentato rubral atrophy; The atrophy of pallidum subthalamic nuclei; Spinobulbar atrophy, optic neuritis, sclerosing panencephalitis (SSPE); Attention deficit disorder; Virus back encephalitis, post poliomyelitis syndrome, Fa Er Cotard; The Joubert syndrome; Guillain-Barre syndrome, agyria, moyamoya; The neuron obstacle of dividing a word with a hyphen at the end of a line; Lonely syndrome, poly glumine is sick, niemann-Pick disease; Progressive multifocal leukoencephalopathy; Pseudotumor cerebri, refsum, Ze Weige syndrome; Supranuclear paralysis; Family ataxia, 2 type spinocebellar ataxias, Rhett syndrome; Shy-Drager syndrome; Tuberous sclerosis, a creutzfeldt jakob disease, chronic fatigue syndrome; Neuropathy comprises that hereditary neuropathy becomes; Diabetic neuropathy and mitosis neuropathy; Neurodegeneration based on Protein virus comprises Creutzfeldt-Jakob disease (CJD); Anomaly CJD; New anomaly CJD; Mad cow disease (BSE); GSS; FFI; Kuru disease and Alper syndrome, Yue Sefushi is sick, acute disseminated encephalomyelitis; Arachnoiditis; Central nervous system's vascular lesion, the forfeiture of limbs function of nervous system, Charcot Marie Tooth; It is sick to restrain cured Bi Shi; Leukodystrophy is prone to suffer from heart failure, asthma; Epilepsy; The auditory nerve degeneration, degeneration of macula, the optic nerve that retinitis pigmentosa and glaucoma cause is degenerated; (b) processing of psychiatric disorders or prevention; Said psychiatric disorders for example is selected from: anxiety disorder (acute stress disorder for example; Panic disorder; Agoraphobia; Social phobia; Specific phobia disease; Obsessive compulsive disorder; Posttraumatic stress disorder; Body dysmorphic disorder and generalization anxiety disorder); Property anxiety disorder (vulvismus for example; Male erectile dysfunction; Male orgasmic disorder and female orgasmic disorder); The childhood period disease (hyperkinetic syndrome (ADHD) for example; A Si Burger syndrome; Autism; Conduct disorder; Oppositional defiant disorder; Separation anxiety sexual disorders and Tu Leiteshi disease); Eating disorders (for example nervous anorexia and bulimia nervosa); The dysthymic disorder is (for example depressed; Serious depressibility obstacle; Bipolar disorder (manic depression); Seasonal affective disorder (SAD); Cyclothymic disorder and dysthymic disorder); Sleep disorder; Cognitive psychiatric disorders (delirium for example; Amnesia); Personality disorder (paranoid personality disorder for example; Schizoid personality disorder; The schizotypal personality disorder; Antisocial personality disorder; Borderline personality disorder; The histrionic personality disorder; Narcissistic personality disorder; Avoidant personality disorder; Dependent personality disorder and obsessive-compulsive personality disorder), mental disorder (schizophrenia for example; Delusional disorder; Brief psychotic disorder; Schizophreniform disorder; Schizoaffective disorder and shared psychotic disorder) and the relevant disease of material (alcohol dependence for example; The dependence of amphetamine-type stimulants; Cannabis relies on; Cocaine relies on; Hallucinogen dependence; Inhalant dependence; Nicotine dependence; Opioid dependence; Phencyclidine dependence and tranquilizer rely on); (c) processing of inflammatory or allergic conditions or prevention, said inflammatory or allergic conditions for example are selected from: cough, pruritus, food intolerance; Psoriasis, croup, irritable bowel syndrome, tinnitus; Prunus mume (sieb.) sieb.et zucc. Ni Ershi disease, the inductive ulcer of the ulcer of stress-induced or aspirin, allergic rhinitis; Allergic dermatitis, conjunctivitis, inflammation; Inflammatory bowel, ileitis, pancreatitis; Cholecystitis, anallergic rhinitis, esophagitis; Osteoarthritis, rheumatoid arthritis, Hay Fever; To the allergic effect reaction of room demodicid mite, to the allergic effect reaction of house pet, Huntington Chorea; Acute inflammation pain, Encelialgia, tooth pain and headache; Inflammatory hyperpathia, sense of touch hyperpathia, allergic skin reaction; The reaction of allergia eye, asthma, atherosclerosis; Arthritis, chronic ulcer (for example relevant chronic angionoma), eczema with rheumatoid arthritis; Keep eupnea; Alleviate throat pain and cough; Normal stool is kept in help; Alleviate stomach discomfort; Help is restored from flu and influenza; As decongestant; Alleviate headache; Alleviate myalgia; Relax slight painful and pain; Alleviation to toothache is provided; Provide the alleviation of oral cavity or gastric ulcer and the joint of keeping health; (d) processing of immune disorders or prevention, said immune disorders for example is selected from: the immunodeficiency patient's condition is AIDS for example, and immunity is patient's condition autoimmune disease such as the systemic lupus erythematosus (sle) (SLE) for example of sexually transmitted disease (STD) condition and impaired immunologic opsonin excessively; (e) processing of tumprigenicity disease or prevention, said tumprigenicity disease for example is selected from: mammary gland, thyroid; Colon, lung, ovary; Skin, muscle, pancreas; Prostate, kidney, genitals; Blood; The unify cancer of skin (like melanoma and Kaposi sarcoma) of immune system (for example spleen, thymus and bone marrow), brain, peripheral nervous system.

4. method according to claim 1; Wherein said method be used for repairing or regenerating nerve unit; Neuronal function or neuroid; Realize regeneration or normalization to neuronic blood flow; The regeneration of injured tissue and healing are for example rebuild after the wound of nerve; Tissue transplantation; In rebuilding after the neural operation; Auxiliary from apoplexy; TIA or other ischemias restore; Auxiliary wound; The healing of bone and muscle; The normalization that neurological's patient's condition or neuron are unusual; Or be used to increase the survival rate of transplanted cells; Increase the fetal cell of the effectiveness of the cell of surviving; Stem cell or other cell therapys, or the method for its combination is united use.

5. method according to claim 1, wherein said method be used to handle or prevent the method for Deviant Behavior or personality trait to unite use.

6. method according to claim 1, wherein said method and wound healing auxiliary are united use.

7. method according to claim 1; Wherein said method be used to improve skin, bone, eye, muscle and other tissue health and for example promote skin from aging, wrinkling or sunlight, wind, rainwater, cold or other damaged the restorative non-Therapeutic Method of influence of the exposure of media; Or for other healthy with safe and comfortable aspects provide comprise muscle and tissue from take exercise, work or consume recovery, the associating use is used in the non-treatment that improves endurance and reduce tired sensation.

8. method according to claim 1, wherein said method be used for handling and prevent normal range and the neurological that be not diagnosable disease of crowd and the non-Therapeutic Method of the psychiatry patient's condition to unite use.

9. according to each described method in the aforementioned claim; Wherein said method is used in the mankind or the animal, and the said mankind or animal are natural overexpression BDNF or GDNF or responsive or interact or the responsive individuality of side effect of the receptor-mediated or enzyme mediation of enzyme interacting medicine to receptor (antagonism) is exciting to the psychiatry side effect of NF simulation or stimulating drug.

10. according to each described method in the aforementioned claim, wherein use said activating agent and do not have the exogenous neurotrophic factor of using.

11. according to each described method in the aforementioned claim, wherein said method is not being used in the situation to the clinic control of curee's the program of using.

12. according to each described method in the aforementioned claim, wherein said activating agent is selected from chinaroot greenbrier sapogenin, smilagenin, table chinaroot greenbrier sapogenin, table smilagenin, zhimusaponin BII, metagenin, samogenin, the cautious ruscogenin of buchu, strange land cautious ruscogenin difficult to understand, texogenin, yonogenin, mexogenin and markogenin and their corresponding esters, ether, ketone and Saponin (glycosylation) derivant.

13. according to each described method in the aforementioned claim, wherein said activating agent is selected from chinaroot greenbrier sapogenin and smilagenin and their corresponding esters, ether, ketone and Saponin (glycosylation) derivant.

14. according to each described method in the aforementioned claim; Wherein said one or more activating agents and one or more adjuvant are united use, and said one or more adjuvant are selected from chemical compound, the energy that can change into the TCA intermediate in metabolism adjuvant, the chemical compound (ketogenic compounds) that increases the ketoboidies level, tricarboxylic acids (TCA) intercycle body, the body and strengthen chemical compound and its any mixture.

15. according to each described method in the aforementioned claim; Wherein said one or more activating agents are used in the compositions that comprises said activating agent and any suitable other compositions, and said compositions is pharmaceutical composition (medicine), food, food supplement or beverage (such as soda pop) or topical composition such as cosmetics, eye or skin (for example dermatological) compositions for example.

16. method according to claim 13, for example medium chain triglyceride (MCT) or medium-chain fatty acid (MCFA) are present in the said compositions to keep dissolving or suspension or the dispersion of said activating agent in said compositions wherein said one or more activating agents with one or more lytic agents and/or suspending agent and/or dispersant.

17. be selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines with and the agent of ester, ether, ketone and glycosylated form, said dose is used for through use one or more said dose of effective dose and the natural NF that under homeostatic control, regulates the curee with avirulent mode to induce the curee method of the stable state of self regulating of NF to the curee.

18. according to claim 17 dose, said dose is used for like each defined method of claim 2 to 16.

19. one kind comprise be selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines with and the compositions of one or more activating agents of ester, ether, ketone and glycosylated form, it is used for inducing in the curee through the natural NF that regulates the curee with avirulent mode to one or more said dose of the effective dose of curee's applying said compositions and under homeostatic control the method for the stable state of self regulating of NF.

20. compositions according to claim 15, said compositions are used for like each defined method of claim 2 to 16.

21. be selected from A/B-cis furostan, furan steroid alkene, spirostane and spirostene steroid sapogenines with and one or more agent of ester, ether, ketone and glycosylated form in the curee, induce the purposes in the medicine of the stable state of self regulating of NF through under homeostatic control, regulating curee's natural NF with avirulent mode in preparation.

22. purposes according to claim 21, wherein said medicine are used for like each defined method of claim 2 to 16.

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