CN107011344A - Method and composition for studying, being imaged and treating pain - Google Patents

Abstract

The application is related to saxitoxin similar compound, composition, pharmaceutical composition, the synthetic method of saxitoxin analog, imaging method, treatment method, and the treatment method includes the method for the treatment of pain.Saxitoxin (STX), gonyatoxin (GTX) and atelopidtoxin and STX variant compounds are combined with sodium channel, and effectively can reduce or block sodium ion to be flowed through from the passage.The blocking of the passage can influence N＆M to act, and effectively can reduce or block sensation of pain, make loosening all muscles, reduction muscle cramp and reduce wrinkle.It may be also useful for positioning, imaging or mark sodium channel that STX analogs are combined with sodium channel, be also useful in research sodium channel and sodium channel disease, and in diagnosing and treating the patient with sodium channel disease therefore.

Description

Method and composition for studying, being imaged and treating pain

It is on May 7th, 2010 applying date that the application, which is, entitled " to be used for the side for studying, being imaged and treating pain The Chinese patent application No.2010800306322 of method and composition " divisional application.

Cross-reference to related applications

According to 35U.S.C. § 119 (e), entitled " the METHODS AND submitted this application claims on May 7th, 2009 COMPOSITIONS FOR STUDYING, IMAGING, AND TREATING PAIN " (are used to studying, be imaged and treating pain The method and composition of pain) U.S. Provisional Patent Application 61/176,172 priority, by reference by its whole During content is incorporated herein.

The application is related to the serial number XX/XXX, XXX that Du Bois et al. submit on May 7th, 2010, entitled " METHODS AND COMPOSITIONS FOR STUDYING, IMAGING, AND TREATING PAIN " (and be used for study, into As and treatment pain method and composition) U.S. patent Nos application, by reference by entire contents simultaneously Enter in the application.

The statement of research is subsidized on federal government

The present invention be governmental support contract/approval number for 5R01NS045684-07 (by NIH Issue) project under make.Government possesses the present invention specific right.

Background technology

" red tide " waters is flooded with noxious material, wherein most notorious is paralytic shellfish poisoning (PSP) (PSP) (referring to text Offer Seafood and Freshwater Toxins：Pharmacology, Physiology, and Detection；Botana, L.M., Ed.；Marcel Dekker：New York, 2000).Form and function all very unique small molecule biguanides base junctions Structure --- saxitoxin, N-STX and gonyatoxin --- represents PSP main component.(referring to forward position skill Art is summarized：(a) Llewellyn, L.E.Nat.Prod.Rep.2006,23,200-222；(b) Hall, S.；Strichartz, G.；Moczydlowski, E.；Ravindran, A.；Reichardt, P.B.ACS Symp.Series 1990,418,29- 65.) these it is highly polar rich in heteroatomic compound be smart design can play obstruction ion stream pass through it is valtage-gated Na⁺Passage (Na_V) effect plug, so as to suppress the electrical conduction in cell.(referring to document：For example, Tetrodotoxin, Saxitoxin, and the Molecular Biology of the Sodium Channel；Eds.C.Y.Kao； S.R.Levinson；Ann.New York Acad.Sci.：New York, Vol 479,1986；Tikhonov, D.B.； Zhorov, B.S.Biophys.J.2005,88,184-197, and references cited therein.)

Complicated molecular shape and these toxin common to these toxin are used as the pharmacology work for studying ion channel The importance of tool, has encouraged people to make great efforts to carry out from the beginning assembling (de novo assembly) to them.Three prior art texts Offer and saxitoxin (STX) and a kind of preparation of deaminizating formyloxy form has been described.(referring to document：For example, (a) Tanino, H.；Nakata, T.；Kaneko, T.；Kishi, Y.J.Am.Chem.Soc.1977,99,2818-2819；(b) Kishi, Y.Heteroeycles 1980,14,1477-1495；(c) Jacobi, P.A.；Martinelli, M.J.；Polanc, S.J.Am.Chem.Soc.1984,106,5594-5598；(d) Martinelli, M.J.；Brownstein, A.D.；Jacobi, P.A.；Polanc, S.Croat.Chem.Acta 1986,59,267-295；(e) Fleming, J.J.；Du Bois, J.J.Am.Chem.Soc.2006,128,3926-3927；(f) Fleming, J.J.；McReynolds, M.D.；Du Bois, J.J.Am.Chem.Soc.2007,129,9964-9975.After their reporting, Kishi etc. describes asymmetric conjunction Into (-)-deaminizating formyl saxitoxin, referring to document：Hong, C.Y.；Kishi, Y.J.Am.Chem.Soc.1992, 114,7001-7006；Referring also to document such as Iwamoto, O.；Koshino, H.；Hashizume, D.；Nagasawa, K.Angew.Chem.Int.Ed.2007,46,8625-8628.)

Pain includes the pain of sharp pain, dull pain, pain and other forms.Pain is that a kind of intensity and duration may Different sensation, and may be caused by a variety of causes.For example, pain is probably acute, such as damage, disease or wound Caused pain；It is probably chronic, pain caused by such as chronic disease or illness, inflammation, cancer or other reasonses；May It is focal or diffused；It is probably low-intensity, moderate strength or high intensity.Therefore, pain is different sensations, bag Include such as Acute Pain, chronic ache, splanchnodynia, surgical pain, arthralgia, ostalgia, backache, headache, neuropathic pain, phantom limb pain And the pain of other various forms and experience.However, in general, regardless of intensity or duration, regardless of For which kind of reason, pain is more preferably mitigated or eliminated in people as far as possible.

Often can by subject's medication come pain of alleviation.It is block nerves signal to mitigate one of mechanism of pain Transmission, is such as realized by block nerves impulsion along the conduction of nerve fibre.For example, by reduction, blocking or can change The activity of sodium channel blocks this nerve impulse.Such as saxitoxin, gonyatoxin, atelopidtoxin and other known to The compound of " 1 sodium channel blockers " etc can influence sodium channel.(referring to document such as Llewellyn, Nat.Prod.Rep.23：200-222(2006)).But as " toxin " one word in its title is implied, this kind of point Son also has the influence in addition to blocking pain.It is desirable to identify effectively mitigate pain without dangerous side-effects Molecule.

Chronic ache is main, universal health problem, and is one of most common reason when people see a doctor." it is What my meeting (chronic) pain" " it is what causes my pain" or " pain from where", these problems are typically to suffer from The individual of chronic ache makes great efforts to obtain the part in the problem of answer.Lack objectively diagnosis detection, limited effective doctor Equipment and safe drugs are treated, certainly make the caregiver of chronic pain patient and they more worried.Suffer from currently used for diagnosis The method of person's pain is very subjective and depends on the self-report of patient.Individual need with chronic ache is stood substantial amounts of Detection, and be often conveyed to receive experience analgesic detection and surgical operation, and this kind for the treatment of is based only upon limited objective base. Further, since the medical imaging technology of the existing level used at present such as X-ray, computed tomography (CT), ultrasonic examination Anatomical abnormalities are largely dependent upon with conventional MRI imaging (MRI), therefore they are in assessment of chronic pain syndrome Aspect still suffers from weak point.Significant interverbebral disc can be found for example with routine MRI in the asymptomatic subjects of 27%-31% It is abnormal.In addition, the nature process of the degenerative disc disease of individual and the no relevance of progress of pain symptom, and pierce Swash property discography and the morphometry based on MR only has faint relevance with backache event.Recently, it has been found that there is disease In shape and the knee of asymptomatic a middle-aged person and the elderly, MRI detects meniscus tear almost equally generally (~60%), this The relevance strengthened again between MR results and pain cause is weaker.However, in spite of this kind of opposite evidence, but unfortunately , for a variety of diseases, these based on anatomical imaging results with the important composition portion for being still treatment protocol Point！Therefore, substantial amounts of patient is by unnecessary operation and unsuitable treatment.

The content of the invention

In short, embodiment of the present invention includes compound, composition, pharmaceutical composition, the method, right for studying pain Method, method for the treatment of pain that pain is imaged etc..

In one embodiment, the invention provides a kind of structure as shown below A, B, C or D compound or its can Medicinal salt, isomer, dynamic isomer or pro-drug.

In embodiments, the invention provides be related to the sufficient toad of saxitoxin (STX), gonyatoxin (GTX) and spot The compound of poison, and STX variant compounds.In embodiments, the STX variant compounds include such conjugates, When being applied to subject, compared with STX, the extended serum half lives of the conjugates, and when being applied to subject, with STX is compared, the acting duration extension of the conjugates.

In one embodiment, the invention provides a kind of method for treating subject, wherein methods described include pair Subject applies the compound of any one structure in having structure A, B, C or D for treating its pain effective dose or its is pharmaceutically acceptable Salt, isomer, dynamic isomer or pro-drug.

In embodiments, should the invention provides a kind of method that subject for making needs receive treatment mitigates pain Method includes applying subject in having structure A, B, C or D of effective dose the compound of any one structure or its is pharmaceutically acceptable Salt, isomer, dynamic isomer or pro-drug, so as to mitigate the pain of the subject.

In one embodiment, the invention provides a kind of voltage-gated sodium channel enhancing activation pain for treating subject The method of pain approach, this method includes applying subject any one in having structure A, B, C or D for treating its pain effective dose Plant the compound or its pharmaceutically useful salt, isomer, dynamic isomer or pro-drug of structure.

In one embodiment, the invention provides a kind of method for preparing saxitoxin analog, including：Make nine The guanidine reaction generation C13-Troc carbonate of yuan of rings；In the presence of a lewis acid, close the guanidine ring in the product of (i)；With And the product of (ii) is aoxidized and is deprotected, so as to generate saxitoxin analog.

In one embodiment, the invention provides a kind of method for preparing saxitoxin analog, including：Make 1- Serine methylester reacts to form aldehyde；The aldehyde of (i) and amine are condensed；The product of (ii) is set to react and effectively ring is closed and generated Carbamide compound；The product of (iii) is set to react in such a process, the process includes allyl deprotection and protected and isothiourea Formation；And by the product amination of (iv), so as to generate saxitoxin analog.

Compound disclosed by the invention can be used for the pain for treating subject, including pain of alleviation, reduction as anodyne The degree and elimination pain and/or pain of pain.

For example they can be used for the treatment of the types of pain of following (and non-limiting) pain to be treated：Acute Pain, anal fissure Pain, arthritis ache, backache, blepharospasm pain, cancer pain, chronic ache, dentistry pain, fibromyalgia, joint Bitterly, neuropathic pain, postoperative pain, friendship after antimigraine, cervical pain, splanchnodynia, neuropathic pain, labor pain, herpes zoster The neural maintenance pain of sense, herpes zoster pain, tension headache, trigeminal neuralgia pain, myositis pain, musculoskeletal pain；Pain in the back, Sprain and pull pain；Integrated with functional intestines problem such as non-ulcer dyspepsia, NCCP and irritable bowel Levy the pain of correlation；The pain related to myocardial ischemia；Toothache；And pain caused by dysmenorrhoea.

Saxitoxin similar compound disclosed herein can be also used for treating following disease and mitigate and these diseases Sick related pain and discomfort：Blepharospasm, arrhythmia cordis, epilepsy, focal dystonia, ephidrosis, muscle cramp and Atony of bladder.

Saxitoxin similar compound disclosed herein can be connected to label, oligonucleotides, protein, lipid, Steroids, antibody or antibody fragment.Label can be for (for example) selected from the group by following material composition：Radio isotope, Fluorescence structure part, chemiluminescent structure part, enzyme, antibody, antibody fragment, magnetic-particle and quantum dot.

Above-mentioned connection can be covalent bond or other connections.Containing with label, oligonucleotides, protein, antibody or antibody The conjugated compound of the saxitoxin similar compound of fragment connection, can be detected, and can application of by analysis It is detected in subject's body of the conjugated compound.It is described containing with oligonucleotides, protein, lipid, steroids, antibody Or antibody fragment connection saxitoxin similar compound conjugated compound, can be directed into required position, organ-tissue, Cell, Cytology Lab.For example, the oligonucleotides of the conjugated compound, protein, lipid, steroids, antibody or antibody fragment knot Saxitoxin analog effectively can be directed to specific voltage-gated sodium channel isoform (isoform) or table by structure part Up to Na_VCell type, or effectively saxitoxin analog can be positioned or anchored near voltage-gated sodium channel.

The pharmaceutical composition of the analog disclosed herein containing saxitoxin includes containing saxitoxin as described herein The pro-drug of any one in similar compound or its isomer, its dynamic isomer or these materials or these The pharmaceutical composition of the pharmaceutically useful salt of any one in material.

Compound and pharmaceutical composition as described herein can be used in the method for the treatment of subject, and methods described is included with energy The amount of subject's illnesses is enough effectively treated, saxitoxin similar compound or its isomerism are applied to subject The drug regimen of any one in the pro-drug or these materials of any one in body, its dynamic isomer or these materials Thing or pharmaceutically useful salt.The disease for example can be pain.Pain disease to be treated includes (for example) Acute Pain, anal fissure pain Bitterly, arthritis ache, backache, blepharospasm pain, cancer pain, chronic ache, dentistry pain, fibromyalgia, arthralgia, Neuropathic pain, postoperative pain, sympathetic god after antimigraine, cervical pain, splanchnodynia, neuropathic pain, labor pain, herpes zoster Through maintenance pain, herpes zoster pain, tension headache, trigeminal neuralgia pain, myositis pain, musculoskeletal pain；Pain in the back, sprain With pull pain；With functional intestines problem such as non-ulcer dyspepsia, NCCP and irritable bowel syndrome phase The pain of pass；The pain related to myocardial ischemia；Toothache；And pain caused by dysmenorrhoea.

Treating the method for subject also includes following methods, and this method includes leading to can effectively treat voltage gated sodium The amount of the enhanced disease in road, saxitoxin similar compound or its isomer, its tautomerism are applied to subject The pharmaceutical composition or pharmaceutically useful of any one in the pro-drug or these materials of any one in body or these materials Salt.In embodiments, the enhanced disease of the voltage-gated sodium channel is selected from the group being made up of following disease：Acute Pain, Anal fissure, arthritis, backache, chronic ache, dentistry pain, fibromyalgia, arthralgia, antimigraine, cervical pain, nerve pain Bitterly, neuropathic pain, postoperative pain, herpes zoster, tension headache or trigeminal neuralgia pain after labor pain, herpes zoster, Blepharospasm, cancer, arrhythmia cordis, epilepsy, focal dystonia, ephidrosis, muscle cramp and atony of bladder.

Treat subject method also include following methods, this method include with can effectively treat following disease, with And mitigate the pain related to these diseases and uncomfortable amount, saxitoxin similar compound is applied to subject or its is same The medicine of any one in the pro-drug or these materials of any one in enantiomers, its dynamic isomer or these materials Compositions or pharmaceutically useful salt, the disease are selected from (for example)：Blepharospasm, arrhythmia cordis, epilepsy, focal Muscle tensility barrier Hinder, ephidrosis, muscle cramp and atony of bladder.

Other method also includes the neuronal activity of reduction subject or causes the method for the loosening all muscles of subject, bag Include can be effectively reduced its neuronal activity or make the amount of its loosening all muscles, it is similar to apply saxitoxin to subject The pro-drug or these materials of any one in compound or its isomer, its dynamic isomer or these materials In any one pharmaceutical composition or pharmaceutically useful salt.

Other method also includes the method for the treatment of subject, and it is included so that the amount of wrinkle can be effectively mitigated or eliminated, Apply any in saxitoxin similar compound or its isomer, its dynamic isomer or these materials to subject The pharmaceutically useful salt of any one in the pro-drug of one or these materials.

Other method also includes the method that is diagnosed to subject, and this method is including with can effectively will be valtage-gated The enhanced disease in sodium channel is positioned at the amount of specific region in subject's body, and the similar chemical combination of saxitoxin is applied to subject Appoint in the pro-drug or these materials of any one in thing or its isomer, its dynamic isomer or these materials The pharmaceutically useful salt for one of anticipating.For example can (such as before or during cat scan process, PET sweeps before or during imaging process Before or during retouching process, before or during MRI processes and before or during SPECT imaging processes), subject is applied Saxitoxin similar compound, it can be the saxitoxin similar compound of mark.

Prepared as shown in Figure 1 there is disclosed herein synthetic method, including by the chemical synthesis process shown in Fig. 4 or Figure 10 The saxitoxin analog naturally occurred method.For example there is disclosed herein the method for preparing gonyatoxin, such as scheme Shown in 10.

Brief Description Of Drawings

Fig. 1 shows saxitoxin analog as described herein (including saxitoxin, the sufficient toad of gonyatoxin and spot Poison) structure.

Fig. 2A shows position of the saxitoxin when being incorporated into sodium channel.

Fig. 2 B further illustrate the diagram for the saxitoxin molecule being in place when being incorporated into sodium channel.

Fig. 3 A show the sodium current (value shown in figure recorded when cell is exposed to the saxitoxin of various concentrations With receive rub (nM) metering).

Fig. 3 B are shown in Chinese hamster ovary celI, and the influence of the saxitoxins of various concentrations to sodium current is with Saxidomus poison The concentration of element and the curve (top curve) that changes and with saxitoxin analog N, N- dimethyl STX (compound 2) Concentration and the curve (bottom curve) that changes.

Fig. 4 is shown available for the synthetic schemes for preparing saxitoxin analog.

Fig. 5 shows saxitoxin analog, and there is provided these analogs confrontation sodium current (rNa_V1.4) effect Value.

Fig. 6 shows the connection strategy (ligation strategy) for modifying saxitoxin analog.

Fig. 7 is shown by being covalently attached the saxitoxin analog to modify from different fluorophors (label) Example.

Fig. 8 shows the saxitoxin analog STX- Malaysias acyl for the tape label for being not present and existing various concentrations Sodium current in the case of imines (compound 19 shown in Fig. 7).

Fig. 9 shows an example of the synthetic schemes of synthesis saxitoxin analog GTX-3 (GTX3).

Figure 10 provides the further diagram of an example of the synthetic schemes to synthesizing saxitoxin analog GTX3 Explanation.

Figure 11 shows the structure of saxitoxin analog GTX3 and GTX-2 (GTX2) and pass therebetween System.

Figure 12 provides the conjunction for being used for synthesizing saxitoxin analog being suitable for imaging research, being connected to label Into an example of scheme.

Figure 13 is provided compared with control rats, and rat is applied by microneedle patch (microneedle patch) Illustrating for the result of comparative study is carried out with STX (0.44 μ g).Compared to control group, the skin handled through STX is to harmful Stimulating the response of (heat) significantly reduces.

Figure 14 A show the experimental result of influence of the STX forms of several C-13 modifications to sodium current (in Chinese hamster ovary celI rNa_V1.4 electric currents).Figure 14 A show received by 0,1 rub (nM), caused by 3nM and 5nM STX sodium current reduction.

Figure 14 B show the figure made with normallized current to Drug level, and it is similar with saxitoxin to be labelled with STX The IC of thing N, N- dimethyl-saxitoxin (compound 10 shown in Figure 14 B)₅₀Value.

Figure 14 C list the IC of several saxitoxin analogs with substituent R as depicted₅₀Value.

Figure 15 shows the method for synthesis fluorescence STX conjugates, and shows the sodium channel being attached in Chinese hamster ovary celI This fluorescence divided.

Figure 16 is shown is injected directly into mouse rear solid end by STX-Cy5 conjugates (shown in red) or STX (shown in blueness) In after carry out mechanical stimulus measured by relative local anaesthesia result.

Figure 17 show N- succinimides 4- [¹⁸F] fluorobenzoate ([¹⁸F] SFV) radio chemistry synthesis and Chemo-selective is connected to NH₂- STX, TTX-s Na are directed to generate_VThe in-vivo imaging medicament of isoform.

Figure 18 shows that left side retains the micro- PET-MRI images (PET of neurotrosis (SNI) rat：Positron emission is broken Layer scanning；MRI：Magnetic resonance imaging).

Figure 19 shows the synthetic schemes of synthesis saxitoxin analog such as gonyatoxin.

Figure 20 shows the synthetic schemes of the STX analogs of synthesis saxitoxin analog such as R7- substitutions.

Figure 21 shows a series of saxitoxin analog of N7- substitutions, and is labelled with IC₅₀Data, the data are Block to measure with sodium current caused by the compound.

Figure 22, which is shown, effectively to provide the synthetic schemes of n-propyl in R3 positions, and further illustrates preparation GTX3 C10- replaces the chemical step of analog.

Detailed description of the invention

Before the present invention is described in more detail, it should be appreciated that present disclosure is not limited to institute The specific embodiment of description, thus, what embodiment of the present invention can certainly change.It is also understood that due to this hair Bright disclosure is only limited by appended claims, thus term as used herein is intended merely to describe specific reality Scheme is applied, without being intended to limit.

Unless otherwise defined, otherwise all technologies used herein and scientific terminology and one of ordinary skill in the art are usual The implication of understanding is identical.

The all publications and patents being related in this specification are hereby incorporated herein by, as specifically designed for every A publication or patent, which are separately indicated, to be incorporated by reference herein equally, and by reference Be incorporated herein is for the disclosure and description method related to cited publication and/or material.

After present disclosure has been read, it will be apparent to one skilled in the art that herein Each embodiment for describing and showing has the composition and feature of separation, is not departing from scope and spirit of the present invention In the case of, it can be easily separated from one another or be combined with the feature of any one in some other embodiments.Described Any method can be to describe the sequencing of event or feasible other are sequentially implemented in logic.

Except as otherwise noted, otherwise the embodiment of the application by using the organic synthesis in the range of art technology , biochemistry, biology, molecular biology, DNA recombinant techniques, pharmacology etc..This kind of technology has abundant theory in the literature It is bright.

List embodiment herein, so as to provided for those skilled in the art how to implement it is described herein and require protect The method of shield and illustrative disclosure and the description for how using compound described herein and claimed.Unless another Indicate outside, otherwise number in terms of parts by weight, temperature by DEG C in terms of, pressure is atmospheric pressure or close to atmospheric pressure.Standard temperature Degree and pressure are defined as 20 DEG C and 1 atmospheric pressure.

Before the embodiments of the present disclosure are described in detail, it should be appreciated that except as otherwise noted, otherwise the present invention not Specific material, reagent, reaction material, preparation process etc. are confined to, thus these can all change.It is also understood that Term as used herein is intended merely to describe specific embodiment, without being intended to limit.In addition it is possible that being disclosed herein The step of it is logically feasible in the case of can carry out in a different order.

It must be noted that in this specification and appended claims, unless context clear stipulaties, otherwise odd number shape Formula " one kind ", " one ", "the" and do not indicate that the mode of quantity covers and has a case that multiple signified things.Thus, for example, When mentioning " compound ", including there are multiple compounds.It will be related in this specification and appended claims And many terms, unless substantially have opposite meaning represents that otherwise these terms are defined as following implication.

Definition

When theme disclosed in the present application is described and claimed as, following term is used according to being defined as below.

Term " saxitoxin analog " includes the sufficient toad of saxitoxin, N-STX, gonyatoxin and spot Malicious AB and related compound.The structure of the related compound will be discussed in further detail below, it should be appreciated that be Including compound shown in Fig. 1 and related compound, the related compound includes the variant related to molecule described herein Compound.As described below, saxitoxin analog is combined with sodium channel." saxitoxin " is abbreviated as " STX ".Knee ditch algae poison Plain is abbreviated as " GTX ".

Term " the saxitoxin analog naturally occurred " refers to saxitoxin, the new Saxidomus found in nature Toxin, gonyatoxin and other related compounds.

Other compounds that can be combined with sodium channel include：Tetraodotoxin (TTX)；And local anesthetic, such as Sai Luoka Because of (xylocaine), Bupivacaine (bupivacaine) and lidocaine (lidocaine).Although most of sodium channels are to river Tetrodotoxin sensitivity (TTX-s), but also have some sodium channels to TTX " insensitive ".

As used herein, " pain " generally refers to physiology and sensation or the impression of psychology of body or physiological pain.Such as this Literary used, " pain " also includes nociception, and it is the life by other aspect mediations of acceptor and neurotransmitter and nervous system The pain of thing impression.Therefore, as used herein, " pain imaging " refers to body or the physiological pain sense for the subject being imaged That receives visually indicates form." pain " can especially be positioned at injury, or be probably systemic；Equally, the figure of pain As can intuitively show the overall status that pain is received, or it can specifically indicate the position or source of pain.Pain includes But it is not limited to：Acute Pain, chronic ache, splanchnodynia, surgical pain, arthralgia, ostalgia, backache, headache, neuropathic pain, The pain of phantom limb pain and other forms.Therefore, pain to be treated includes but is not limited to：Acute Pain, anal fissure pain, joint Scorching pain, backache, blepharospasm pain, cancer pain, chronic ache, dentistry pain, fibromyalgia, arthralgia, antimigraine, Neuropathic pain, postoperative pain, sympathetic nerve are maintained after cervical pain, splanchnodynia, neuropathic pain, labor pain, herpes zoster Property pain, herpes zoster pain, tension headache, trigeminal neuralgia pain, myositis pain, musculoskeletal pain；Pain in the back, sprain and pull Pain；To functional intestines problem such as non-ulcer dyspepsia, the NCCP pain related with irritable bowel syndrome Bitterly；The pain related to myocardial ischemia；Toothache；And pain caused by dysmenorrhoea.

Saxitoxin similar compound disclosed herein can also be used for treatment blepharospasm, cancer, arrhythmia cordis, insane Epilepsy, focal dystonia, ephidrosis, muscle cramp and atony of bladder, and mitigate the pain related to these diseases and It is uncomfortable.

Neuropathic pain syndrome is traditionally according to facilitating the disease or situation of the syndrome to classify.Nerve is ached Pain syndrome includes：Diabetic neuropathy；Sciatica；Nonspecific low back pain；Multiple sclerosis pain；Fibromyalgia； The DPN related to AIDS；PHN；Trigeminal neuralgia；And by physical trauma, amputation, cancer, The pain that toxin or chronic inflammatory condition are caused.These situations are difficult to treat, although and known to have several drugses to have limited Curative effect, but seldom can completely control pain.The symptom of neuropathic pain has incredible heterogeneity, and past It is past to be described as spontaneous acutely shouting pain, or persistent burning pain.In addition, also having the pain related to conventional non-sensation of pain, example Such as, " on tenterhooks " (cacesthesia and insensitive), the sensitiveness enhancing (hyperesthesia) to touch, after fanout free region sexual stimulus The pain (dynamic and static or heat anomaly pain) of generation, the sensitiveness to noxious stimulation strengthen (hot, cold, mechanicalness pain sensation mistake It is quick), remove the sensation approach missing of post-stimulatory constant pain sense (hyperalgia) or selectivity or not enough (pain sensation subtracts Move back).

As used herein, term " therapy ", " treatment " and " treatment " is defined as using compound disclosed in the present application or group Compound mitigates or improved pain and/or generation, sensation, perception and/or the influence of its symptom.As used herein, " therapy " covers Any therapy for the treatment of main body (such as mammal, the non-human animal generally paid close attention to for people or animal doctor) pain, and including： (a) possibility that pain occurs for subject is reduced, (b) hinders the startup of pain, and (c) to mitigate pain, i.e. make pain regression And/or alleviate one or more pain symptoms.Therefore, the statement of term " therapy ", " treatment " and " treatment " and the like Including：Pain of alleviation, the pain that mitigates, limitation pain, reduction pain, analgesia, improvement pain, blocking pain, inhibition of pain, and Other act on sensation of pain, pain perceive, propagation of the pain signal in nervous system or other act on the row of pain For mode.

As used herein, term " preventative treatment " or " prophylactic treatment " refer to completely or partially inhibition of pain or its Symptom, and/or can play treatment in partially or completely pain of alleviation and/or in terms of mitigating the adverse effect as caused by pain Effect.

As used herein, term " main body ", " subject " or " patient " include people and mammal (such as mouse, rat, Pig, cat, dog and horse).The typical body of compound disclosed in the present application can be applied for mammal, particularly primate, The especially mankind.For veterinary application, it is all suitable to have diversified subject, is, for example,：Livestock, such as ox, sheep, Goat, milk cow, pig etc.；Poultry, such as chicken, duck, goose, turkey；And domestic animal, particularly pet, such as dog and cat.It is medical For disconnected or research application, it is all suitable subject to have diversified mammal, including rodent is (such as mouse, big Mouse, hamster), rabbit, primate and pig (such as inbred pig).Term " living main body " refer to main body mentioned above or its His lived organism.Term " main body living " refers to whole main body or organism, rather than one only cut from main body living Divide (such as liver or other organs).

Term " compound of separation " is separated in referring to generally naturally occur other compounds of when institute's association from it Out, or relative to other compounds of association the compound being enriched with.The compound purity of separation is generally at least about 80 weights Measure %, at least about 90 weight %, at least about 98 weight % or at least about 99 weight %.Disclosure is intended to include diastereomeric different The pure form and its pharmaceutically useful salt for the enantiomerism that structure body and their racemic form and fractionation are obtained.

As used herein, term " unit dosage forms " refers to be suitable as the thing of the unit dose for people and/or animal subjects Manage separation unit, each unit comprising scheduled volume compound and therewith associated with pharmaceutically useful diluent, carrier or tax Shape agent, the scheduled volume is to be enough to produce the amount of intended effect by calculating.The specification of unit dosage forms depends on the specific of use Compound, method of administration and frequency, the effect to be reached, and pharmacodynamics of each compound in main body.

" pharmaceutically useful excipient ", " pharmaceutically useful diluent ", " pharmaceutically useful carrier " or " pharmaceutically useful adjuvant " refers to Such excipient, diluent, carrier and/or adjuvant, its can be used for preparing it is generally safe and nontoxic and without biology or The pharmaceutical composition of other undesirable situations of aspect, and including can be excipient for animals and/or human pharmaceutical use, diluent, Carrier and adjuvant.In the specification and claims " pharmaceutically useful excipient, diluent, carrier and/or adjuvant " used Include the situation of excipient as one or more, diluent, carrier and adjuvant.

As used herein, " pharmaceutical composition " is intended to include being adapted to be applied to subject (such as mammal, particularly people) Composition." pharmaceutical composition " is usually sterile, and preferably without those subject can be caused to produce adverse reaction Pollutant (for example, the compound in pharmaceutical composition is pharmaceutical grade).Pharmaceutical composition can be designed to be able to by it is a variety of not With method of administration be applied to and need the subject or patient of the pharmaceutical composition, method of administration include it is oral, intravenous, contain Clothes, rectum, parenteral, intraperitoneal, intracutaneous, tracheal strips, intramuscular, subcutaneous, suction etc..

As used herein, term " therapeutically effective amount " and " effective amount " alternatively use, refer to it is being applied, be enough to make Compound (alternatively referred to as medicament, medical compounds or medicine, and be contained in composition or medicine group that intended application comes into force In compound) amount, the intended application includes but is not limited to treatment pain or treats disease or illness.For example, effective dose Compound can alleviate one or more symptoms of treated pain, illness or disease to a certain extent, and/or can be Suppress to a certain extent the existing positive main body for receiving treatment or one kind of pain, illness or disease for possibly producing or A variety of symptoms.Therapeutically effective amount according to intended application (external or in vivo) or subject's (body weight of such as subject and age), with And pain, illness or disease (such as light and heavy degree of pain, illness or disease), the method for administration treated can be different, Those skilled in the art can readily determine that therapeutically effective amount.The term is also applied for inducing the particular responses of target cell Dosage.According to selected particular compound, use administering mode, whether with other compound administering drug combinations, administration time, give Medicine tissue and the physics delivery system for carrying compound, specific dosage can be different.

Herein, term " serum half-life " and " plasma half-life " use as is understood in the art, and Refer to that amount or concentration of the material in the serum of subject drop to time used in the half of initial value after administration.Serum half Decline phase and plasma half-life can be used for determining or infer compound (such as medical compounds, tracer or other can be applied to it is tested The compound of person) acting duration；Longer serum half-life and longer plasma half-life show the compound tool There is longer acting duration.

As used herein, term " acting duration " refers to after administration, and the compound produces aobvious to the subject of medication Write the time duration of effect.For example, in the case where compound has anesthetic effect, acting duration, which includes applying, to be somebody's turn to do Subject undergoes the time of anesthetic effect after compound.The compound than reference compound with longer acting duration is Such compound, the effect (as anaesthetized) of the compound is longer than effect (as anaesthetized) duration of reference compound.

As used herein, " sodium channel " is any one in a major class macromolecular, and it is naturally occurring in biomembrane (such as god Through cell membrane) in.When being present in the film that nature just has, sodium channel is sensitive to the voltage differences of these film both sides, and can be with Sodium ion is allowed to pass through cell membrane.Be combined with some toxin, can block sodium ion by (otherwise, if the not no toxin, Then sodium ion will pass through).Spontaneous sodium channel has diversified forms (such as to identify coding in mammalian cell Ten kinds of unique Na⁺Passage isoform (Na_V1.1-1.9, Na_X, wherein x refers to sodium channel hypotype) gene (referring to document Hille, B.Ion Channels of Excitable Membranes, the third edition, Sinauer：Sunderland, MA, 2001. 73- Page 78).Term " sodium channel " used herein includes sodium channel variant, and either genetic mutation, splice variant, glycosylation becomes Body, post translational processing variant or other variants (including artificial variants).

Sodium channel on film can transmit charged ion (particularly sodium ion), and can produce can be by various experimental techniques " sodium current " of measurement.The amplitude and time course of sodium current can by external factor (including apply medicine and toxin) shadow Ring.STX compounds (including saxitoxin analog disclosed herein) can reduce the amplitude of sodium current.This reduction can be described as " blocking " or " obstruction " or the term of other expression amplitude reductions, and partially or completely ion can be blocked to be flowed through from sodium channel.

Sodium channel is the key that N＆M system normally functions.The blocking of sodium current (such as passes through Saxidomus poison Plain analog is combined to block with sodium channel) energy block nerves conduction, as blocked along pain sensation fiber and other nerve fibres Conduction, prevention contraction of muscle, therefore sensation, motion and other physiological attributes of subject can be influenceed.The resistance of pain sensation fiber Plug can cause analgesia and anaesthesia；The blocking of muscular movement can cause reduction or the paralysis of impacted muscle, and energy Cause the relaxation of impacted muscle.The relexation is effectively reduced or eliminated wrinkle of skin, wherein impacted muscle bag Include the muscle near skin or be connected to the muscle of skin.The blocking of sodium current (is such as led to by saxitoxin analog and sodium Road combines to block) cardiovascular system, gastrointestinal system, bladder, heart, sense organ and other organs and organ system can be influenceed System.Subject, which is applied, effectively can reduce or eliminate its sodium channel activity (either in specific location, or at a certain group Knit, in organ, tract, or whole body) saxitoxin analog, be beneficial to treat or improve disease, imbalance and Illness and the mitigation pain and discomfort related to these imbalances, including pain, muscle cramp, epilepsy, blepharospasm, office Stove myodystony, ephidrosis and atony of bladder, cardiac disorder and illness (including arrhythmia cordis), alimentary canal imbalance and disease Disease, sensory disorder and illness, cancer, skin disorder, other imbalances disclosed herein and illness.

As used herein, " acceptor " refers to the molecule of binding partner or a part for molecule；For example lead to when STX is attached to sodium During road, sodium channel is used as STX acceptors.Different molecules is combined with different specificity with acceptor；Some parts are with high special Property is combined with acceptor.

As used herein, term " part " refers to the molecule for being attached to acceptor；In embodiments, part may be than it With reference to acceptor be more easy to movement.In embodiments, part can be smaller than the acceptor of its combination or lower point of quality Son.For example STX molecules generally, quality smaller than sodium channel is lower in physiological conditions and is more easy to movement.

As used herein, term " receptor-specific " refers to the specificity that particular ligand is combined with acceptor.Receptor-specific Measurement result provide interphase interaction for acceptor and part pharmacokinetic properties understanding and details.

" pharmaceutically useful salt " refers to the biological efficacy for keeping standard items (free base) and other optional characteristics Those salt (organic salt or inorganic salts).Therefore, " pharmaceutically useful salt " includes any pharmaceutically acceptable and with expected pharmacological property Salt." pharmaceutically useful salt " can be derived from inorganic or organic acid or inorganic or organic base any salt.Term is " pharmaceutically useful Anion " refers to the anion of the acid-addition salts.Term " pharmaceutically useful cation " refers to the cation as formed by alkali addition. The salt and/or anion or cation are selected as not having disadvantage in terms of biology or other.Pharmaceutically useful salt can lead to Cross and obtained with inorganic or organic acid reaction, described acid is, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, second Sulfonic acid, p-methyl benzenesulfonic acid, salicylic acid, malic acid, maleic acid, butanedioic acid, tartaric acid, citric acid etc..Such salt includes deriving Dysgenic inorganic or organic acid or inorganic or organic base (including amino acid) will not be also produced in any way from nontoxic Salt.Suitable inorganic salts include with alkali metal (such as sodium, potassium, magnesium, calcium and aluminium) formation salt.Suitable organic salt includes With organic base (alkali of such as amine, such as monoethanolamine, diethanol amine, triethanolamine, tromethamine, N- methylglucamines) formation Those salt.Described salt also include with inorganic acid (such as hydrochloric acid and hydrobromic acid) and organic acid (such as acetic acid, citric acid, maleic acid, And alkane sulfonic acid and arene-sulfonic acids, such as methanesulfonic acid and benzene sulfonic acid) formed acid-addition salts.

" pharmaceutically useful ester " refers to any esters pharmaceutically useful and with desired pharmacological characteristics.Pharmaceutically useful ester includes The ester that the carboxyl present in compound, sulfonyloxy and phosphonato group are formed, such as C_1-6Arrcostab.When in the presence of two acid During property group, pharmaceutically useful salt or ester can be mono-acid mono-salt or mono-acid monoesters or be disalt or diester；Similarly, presence is worked as During more than two acidic-groups, part or all of acidic-group can be into salt or esterification.

In the case of disclosed particular compound forming salt, these salt are also in the range of disclosure.Unless It is otherwise noted, the compound for otherwise mentioning this paper arbitrary structures formulas should be read to include and mention its salt.Term used herein " salt " refers to the acid salt and/or basic salt with the formation of inorganic and/or organic bronsted lowry acids and bases bronsted lowry.In addition, when compound is simultaneously comprising alkali When property structure division and acid structure part, amphion (" inner salt ") can be formed, this is also included within term used herein In the range of " salt ".It is preferred that pharmaceutically useful salt (such as nontoxic, physiologically acceptable salt), but other salt be also it is useful, For example used in the isolated or purified step that preparation process may be used.The salt of compound for example can shape in the following manner Into：All if in making the medium that salts out or in an aqueous medium, make the compound and a certain amount of acid or alkali (for example Equivalent) reaction, then freeze.

Medicament comprising basic moieties can be with various organic or inorganic acid forming salts.Exemplary acid-addition salts include Acetate (such as with acetic acid or the salt of three halogen acetic acids (such as trifluoroacetic acid) formation), adipate, alginates, ascorbate, day Winter propylhomoserin salt, benzoate, benzene sulfonate, disulfate, borate, butyrate, citrate, camphor salt (camphorate), camsilate, cyclopentane propionate, digluconate, lauryl sulfate, esilate, prolong recklessly Rope hydrochlorate, glucose enanthate, glycerophosphate, Hemisulphate, enanthate, caproate, hydrochloride (salt with hydrochloric acid formation), hydrogen Bromate (salt with hydrogen bromide formation), hydriodate, 2- isethionates, lactate, malate are (with malic acid formation Salt), maleate (salt with maleic acid formation), esilate (salt with ethyl sulfonic acid formation), mesylate is (with methanesulfonic acid shape Into salt), 2- naphthalene sulfonates, nicotinate, nitrate, oxalates, pectate, persulfate, 3- phenylpropionic acids salt, phosphate (salt with phosphoric acid formation), picrate, pivalate, propionate, salicylate, succinate, sulfate are (such as with sulfuric acid The salt of formation), sulfonate (as mentioned in this article those, including the salt formed with p-methyl benzenesulfonic acid), tartrate, thiocyanic acid Salt, toluene fulfonate such as tosilate, undecylate etc..

In the embodiment with feature disclosed in the present application, saxitoxin analog may include or can be formed Salt, including：Chloride, acetate, ascorbate, aspartate, benzoate, benzene sulfonate, disulfate, boric acid Salt, gluconate, citrate, sulfate, tartrate and toluene fulfonate.

Compound containing acid structure part can be with various organic or inorganic alkali forming salts.Exemplary basic salt includes Ammonium salt, alkali metal salt (such as sodium, lithium and sylvite), alkali salt (such as calcium and magnesium salts), with organic base formation salt it is (described organic Alkali is, for example, organic amine, and such as tardocillin, dicyclohexylamine, hydrabamine phenoxy methylpenicillin are (by N, N- pairs of (dehydrogenated rosin) ethylenediamine shapes Into), N- methyl-D-glucosamines, N- methyl-D-glucamides, tert-butylamine) and with amino acid (such as arginine, lysine) The salt of formation.

Nitrogen-containing basic group can be quaternized by following compounds, and the compound is, for example, elementary alkyl halide (such as first Base, ethyl, propyl group and butyl chloride compound and its bromide and iodide), dialkyl sulfate (such as dimethyl, diethyl, two Butyl and diamyl sulfate), long-chain halogenated hydrocarbons (such as decyl, lauryl, nutmeg base and stearyl chlorides and its bromination Thing and iodide), aralkyl halide (such as benzyl bromine and phenethyl bromide).Chemical combination disclosed in the present application is additionally contemplates that herein The solvate of thing.

In one embodiment, compound disclosed in the present application can have one or more chiral centres, thus can quilt Single stereoisomer or stereoisomer mixture is made, it is to use single stereoisomer that this, which depends on original material, Still stereoisomer mixture is used.Unless otherwise stated, the description or name of compound or compound group are intended to together When include single isomers or stereoisomer mixture (racemic or other).Determine that spatial chemistry attribute and separation are three-dimensional The method of isomers is known to those skilled in the art [referring to document March J.：Advanced Organic The discussion of 4th chapter in Chemistry (the 4th edition, John Wiley and Sons, New York, N.Y., 1992)].

Disclosed reactive compound and its salt can in their tautomeric form in the presence of, it is all such Tautomeric forms are also contemplated for the part for present disclosure.

All stereoisomers of compound due to the asymmetric carbon atom on various substituents (for example, there may be Those stereoisomers；Including enantiomeric form (also may be present even if no asymmetric carbon atom) and diastereo-isomerism shape Formula) it is taken into account in the range of disclosure.The independent stereoisomer of compound disclosed in the present application can (for example) Be substantially free of other isomers, or can mix (such as) turn into racemate with it is every other or other selected by Stereoisomer is mixed.The Stereocenter of compound disclosed in the present application can have such as IUPAC 1974Recommendations Defined S or R configurations.

Term " pro-drug " refers to the nonactive precursor of compound, and it is converted into biologically active form in vivo.Precursor Medicine is typically beneficial, and this is due to that pro-drug is easier administration than parent compound in some cases.Pro-drug It is for example when can be oral available for biology, and parent compound is then really not so.In pharmaceutical composition, pro-drug It is also possible to that there is more preferable dissolubility than parent drug.Pro-drug can be converted into parent drug, the machine by various mechanism System includes enzymatic processes and metabolism is hydrolyzed.Referring to documents below：Harper, N.J. (1962) .Drug Latentiation in Jucker, ed.Progress in Drug Research, 4：221-294；Morozowich etc., (1977) .Application of Physical Organic Principles to Prodrug Design in E.B.Roche ed.Design of Biopharmaceutical Properties through Prodrugs and Analogs, APhA； Acad.Pharm.Sci.；E.B.Roche, ed. (1977) .Bioreversible Carriers in Drug in Drug Design, Theory and Application, APhA；H.Bundgaard, ed. (1985) Design of Prodrugs, Elsevier；Wang etc., (1999) Prodrug approaches to the improved delivery of peptide Drug, Curr.Pharm.Design.5 (4)：265-287；Pauletti etc., (1997) .Improvement in peptide bioavailability：Peptidomimetics and Prodrug Strategies, Adv.Drug.Delivery Rev.27：235-256；Mizen etc., (1998) .The Use of Esters as Prodrugs for Oral Delivery Of β-Lactam antibiotics, Pharm.Biotech.11,：345-365；Gaignault etc., (1996) .Designing Prodrugs and Bioprecursors I.Carrier Prodrugs, Pract.Med.Chem.671-696； M.Asgharnejad (2000) .Improving Oral Drug Transport Via Prodrugs, in G.L.Amidon, P.I.Lee and E.M.Topp, Eds., Transport Processes in Pharmaceutical Systems, Marcell Dekker, p.185-218；Balant etc., (1990) Prodrugs for the improvement of drug Absorption via different routes of administration, Eur.J.Drug Metab.Pharmacokinet., 15 (2)：143-53；Balimane and Sinko(1999).Involvement of Multiple transporters in the oral absorption of nucleoside analogue, Adv.Drug Delivery Rev., 39 (1-3)：183-209；Browne (1997) .Fosphenytoin (Cerebyx), Clin.Neuropharmacol.20(1)：1-12；Bundgaard(1979).Bioreversible derivatization of drugs--principle and applicability to improve the therapeutic effects of Drugs, Arch.Pharm.Chemi.86 (1)：1-39；H.Bundgaard, ed. (1985) Design of Prodrugs, New York：Elsevier；Fleisher etc., (1996) .Improved oral drug delivery：solubility Limitations overcome by the use of prodrugs, Adv.Drug Delivery Rev.19 (2)：115- 130；Fleisher etc., (1985) .Design of prodrugs for improved gastrointestinal Absorption by intestinal enzyme targeting, Methods Enzymol.112：360-81；Farquhar D etc., (1983) .Biologically Reversible Phosphate-Protective Groups, J.Pharm.Sci., 72(3)：324-325；Han, H.K. etc., (2000) .Targeted prodrug design to optimize drug Delivery, AAPS PharmSci., 2 (1)：E6；Sadzuka Y.(2000).Effective prodrug liposome And conversion to active metabolite, Curr.Drug Metab., 1 (1)：31-48；D.M.Lambert (2000) Rationale and applications of lipids as prodrug carriers, Eur.J.Pharm.Sci., 11Suppl 2：S15-27；Wang, W. etc., (1999) Prodrug approaches to the Improved delivery of peptide drugs.Curr.Pharm.Des., 5 (4)：265-87.

As used herein, term " substitution " preferably refers to：Replaced with a kind of specified substituent or a variety of substituents, can permitted The substitution of many kinds of degree, unless otherwise indicated.

As used herein, term " optional " refers to that situation about then describing can occur to occur, and both includes The situation of generation, the also situation including not occurring.

As used herein, term " derivative " and its form of presentation grammatically deformed preferably refer to disclosed in the present applicationization Any chemical derivative of compound, such as ester or acid amides, and be preferably the compound with pharmacy function, it is being applied to the food in one's mouth Can (direct or indirect) offer compound disclosed in the present application or its active metabolite during newborn animal.Such derivative is ability Field technique personnel need not move through excessive experiment and just can be clear from, and refer to document：Burger′s Medicinal Chemistry And Drug Discovery, 5th Edition, Vol 1：Principles and Practice are (to quote The mode content of physiological functional derivative of being instructed the document be incorporated into herein).Such derivative includes So-called prodrug compound, such as through alkyl, acyl group, sugar or peptide (such as oligopeptides) derivatization and easily degrade or be metabolized For reactive compound disclosed in the present application those according to compound disclosed in the present application.It is public that such derivative includes the application The Biodegradable polymer derivative for the compound opened.Suitable polymer and prepare Biodegradable polymer derivative Method is known in the art, for example, see document：Int.J.Pharm.115,61-67 (1995).

Herein, term " lipid " is used as is understood in the art, and refers to a major class chemical combination Thing, it includes but is not limited to (such as) sterol, monoglyceride, diglyceride, triglycerides, phosphatide, sphingolipid, polyketide Thing, fat and wax.

Herein, term " steroids " is used as is understood in the art, and refers to a major class chemicals, One of illustrative examples are cholesterol, and it includes one and includes four fused rings (three cyclohexane rings and a pentamethylene Ring) gonane parent nucleus, in addition to the compound with the ring structure aoxidized in various degree, and be included on ring structure there is institute The compound of possible substituent.

Term " administration " refers to compound disclosed in the present application being introduced into main body.A kind of preferred method of administration is oral Administration.Another preferred approach is intravenous administration.But any method of administration can be used, for example locally, subcutaneously, it is peritonaeum, dynamic In arteries and veins, suction, vagina, rectum, nasal cavity, it is introduced into cerebrospinal fluid or to be instilled into body cavity (body compartment) interior.

Term " aliphatic group " refers to saturation or undersaturated straight or branched hydrocarbyl group, including such as alkyl, alkenyl and Alkynyl group.

Term " alkane " or " alkyl " refer to such straight or branched hydrocarbyl group, and it has 1 to 24 carbon atom or had 1 to 12 carbon atom with 2 to 5 carbon atoms or with 6 to 18 carbon atoms or preferably has 2 to 12 carbon atoms, Such as methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, the tert-butyl group, amyl group, hexyl, heptyl, n-octyl, dodecane Base, octadecyl, amyl group, 2- ethylhexyls etc..Unless otherwise indicated, otherwise alkyl group is optionally selected from by one or more Following substituent group：Aryl (optionally substituted), heterocycle (optionally substituted), carbocyclic ring (optionally substituted), halogen, hydroxyl, Shielded hydroxyl, alkoxy (such as C₁To C₇) (optionally substituted), acyl group (such as C₁To C₇), aryloxy group (such as C₁To C₇) (optionally Substitution), Arrcostab (optionally substituted), aryl ester (optionally substituted), alkanoyl (optionally substituted), aroyl (optionally Substitution), carboxyl, shielded carboxyl, cyano group, nitro, amino, substituted-amino, (monosubstituted) amino, (disubstituted) amino, Shielded amino, acylamino-, carbamate, lactams, urea, urethane, sulfonyl etc..

Term " alkenyl " refers to such straight or branched hydrocarbyl group, and it has 2 to 12 carbon atoms or with 2 to 4 Carbon atom with 2 to 5 carbon atoms or with 6 to 18 carbon atoms or preferably has 2 to 12 carbon atoms, and has There are at least one C=C double bonds (cis or trans), such as vinyl.Unless otherwise indicated, otherwise alkenyl group optionally by one It is individual or multiple selected from following substituent group：Aryl (including substituted aryl), heterocycle (including substituted heterocycle), carbocyclic ring (including take For carbocyclic ring), halogen, hydroxyl, alkoxy (optionally substituted), aryloxy group (optionally substituted), Arrcostab (optionally substituted), virtue Base ester (optionally substituted), alkanoyl (optionally substituted), aroyl (optionally substituted), cyano group, nitro, amino, substitution ammonia Base, acylamino-, carbamate, lactams, urea, urethane, sulfonyl etc..

Term " alkynyl " refers to such straight or branched hydrocarbyl group, and it has 2 to 12 carbon atoms or with 2 to 4 Carbon atom with 2 to 5 carbon atoms or with 6 to 18 carbon atoms or preferably has 2 to 12 carbon atoms, and has There are at least one keys of C ≡ C tri-, such as acetenyl.Unless otherwise indicated, otherwise alkynyl group is optionally selected from by one or more Following substituent group：Aryl (including substituted aryl), heterocycle (including substituted heterocycle), carbocyclic ring (including substitution carbocyclic ring), halogen, Hydroxyl, alkoxy (optionally substituted), aryloxy group (optionally substituted), Arrcostab (optionally substituted), aryl ester are (optionally substituted ), alkanoyl (optionally substituted), aroyl (optionally substituted), cyano group, nitro, amino, substituted-amino, acylamino-, amino Formic acid esters, lactams, urea, urethane, sulfonyl etc..

Term " alkoxy " refers to the alkyl group for being connected to oxygen, i.e. R-O-.R represents alkyl group in the formula.One Example is methoxy group CH₃O-。

" organic group " can for functionalization or include additional functionality (such as carboxyl, ammonia being connected with the organic group Base, hydroxyl etc.), it can be shielded or not protected.Satisfy for example, term " alkyl group " not only includes pure open chain With hydrocarbon alkyl substituent, such as methyl, ethyl, propyl group, the tert-butyl group, in addition to other substituents known in the art The alkyl of (such as hydroxyl, alkoxy, alkyl sulphonyl, halogen atom, cyano group, nitro, amino, carbamate, carboxyl) takes Dai Ji.Therefore, " alkyl group " includes ether, ester, haloalkyl, 4-nitro alkyl, carboxyalkyl, hydroxyalkyl, sulfoalkyl etc..

" cyano group " refers to-CN functional groups.

Term " halogen " and " halogen " refer to fluorine, chlorine, bromine or iodine base.There can be one or more (identical or different) halogens Atom.

Term " haloalkyl " refers to the alkyl defined above replaced by one or more halogen atoms.The halogen atom It may be the same or different.Term " dihalo alkyl " refers to by two alkane defined above that can be replaced with identical or different halogen Base.Term " tri haloalkyl " refers to by three alkyl defined above that can be replaced with identical or different halogen.Term is " complete Haloalkyl " refers to the haloalkyl defined above that each hydrogen atom in alkyl group is substituted with halogen atoms.Term " perfluoroalkyl " refers to each hydrogen atom in alkyl group by fluorine-based substituted haloalkyl defined above.

Term " cycloalkyl " refers to full saturation or part is undersaturated single, double or three ring filling rings.The example bag of the group Include cyclopropyl, cyclobutyl, cyclopenta, cyclohexyl, suberyl, adamantyl, cyclooctyl, cis or trans naphthalane, two rings [2.2.1] hept-2-ene", hexamethylene -1- alkenyls, the amyl- 1- alkenyls of ring, Isosorbide-5-Nitrae-cyclo-octadiene base etc..Unless otherwise indicated, otherwise cycloalkanes Base group is optionally selected from following substituent group by one or more：Aryl (including substituted aryl), heterocycle (including replace it is miscellaneous Ring), carbocyclic ring (including substitution carbocyclic ring), halogen, hydroxyl, shielded hydroxyl, alkoxy (such as C₁To C₇) (optionally substituted), acyl Base (such as C₁To C₇), aryloxy group (such as C₁To C₇) (optionally substituted), Arrcostab (optionally substituted), aryl ester be (optionally substituted ), alkanoyl (optionally substituted), aroyl (optionally substituted), carboxyl, shielded carboxyl, cyano group, nitro, amino, take For amino, (monosubstituted) amino, (disubstituted) amino, shielded amino, acylamino-, carbamate, lactams, urea, urine Alkane, sulfonyl etc..

Term " (cycloalkyl) alkyl " refers to the group of naphthene base defined above replaced by alkyl group defined above. The example of the group include (cyclohexyl) methyl, 3- (cyclopropyl)-n-propyl, 5- (cyclopenta) hexyl, 6- (adamantyl) oneself Base etc..Unless otherwise indicated, otherwise (cycloalkyl) alkyl group is optionally selected from following substituent group by one or more：Alkane Base (including substitution alkyl), aryl (including substituted aryl), heterocycle (including substituted heterocycle), carbocyclic ring (including substitution carbocyclic ring), halogen Element, hydroxyl, shielded hydroxyl, alkoxy (such as C₁To C₇) (optionally substituted), acyl group (such as C₁To C₇), aryloxy group (such as C₁Extremely C₇) (optionally substituted), Arrcostab (optionally substituted), aryl ester (optionally substituted), alkanoyl (optionally substituted), fragrant acyl It is base (optionally substituted), carboxyl, shielded carboxyl, cyano group, nitro, amino, substituted-amino, (monosubstituted) amino, (double to take Generation) amino, shielded amino, acylamino-, carbamate, lactams, urea, urethane, sulfonyl etc..

Term " substituted-phenyl " refers to by one or more (being in some cases one, two or three) selected from following The phenyl group that is replaced of structure division：Halogen, hydroxyl, shielded hydroxyl, cyano group, nitro, trifluoromethyl, C₁To C₇Alkane Base, C₁To C₇Alkoxy, C₁To C₇Acyl group, C₁To C₇It is acyloxy, carboxyl, oxygen carboxyl, shielded carboxyl, carboxymethyl, protected Carboxymethyl, methylol, shielded methylol, amino, shielded amino, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, formamide, shielded formamide, N- (C₁To C₆Alkyl) formamide, shielded N- (C₁ To C₆Alkyl) formamide, the N, (C of N- bis-₁To C₆Alkyl) formamide, trifluoromethyl, N- ((C₁To C₆Alkyl) sulfonyl) amino, N- (benzenesulfonyl) amino or phenyl (substituted or unsubstituted, so as to form such as xenyl or naphthyl group).

The example of term " substituted-phenyl " includes：Single (halo) or two (halo) phenyl groups, such as 2,3 or 4- chlorphenyls, 2,6- dichlorophenyls, 2,5- dichlorophenyls, 3,4- dichlorophenyls, 2,3 or 4- bromophenyls, 3,4- dibromo phenyls, the chloro- 4- fluorobenzene of 3- Base, 2,3 or 4- fluorophenyls etc.；Single (hydroxyl) or two (hydroxyl) phenyl groups, such as 2,3 or 4- hydroxyphenyls, 2,4- dihydroxyphenyls, its Shielded hydroxy derivatives etc.；Nitrophenyl group, such as 2,3 or 4- nitrobenzophenones；Cyanophenyl group, such as 2,3 or 4- nitriles Base phenyl；Single (alkyl) or two (alkyl) phenyl groups, such as 2,3 or 4- tolyls, 2,4- 3,5-dimethylphenyls, 2,3 or 4- (isopropyls Base) phenyl, 2,3 or 4- ethylphenyls, 2,3 or 4- (n-propyl) phenyl etc.；Single (alkoxy) or two (alkoxy) phenyl groups, Such as 2,6- Dimethoxyphenyls, 2,3 or 4- (isopropoxy) phenyl, 2,3 or 4- (tert-butoxy) phenyl, 3- ethyoxyl -4- first Phenyl etc.；2,3 or 4- trifluoromethyls；Mono carboxylic or dicarboxyphenyi or (shielded carboxyl) phenyl group, such as 2,3 or 4- carboxyl phenyls or 2,4- bis- (shielded carboxyl) phenyl；Single (methylol) or two (methylol) phenyl or (protected Methylol) phenyl, such as 2,3 or 4- (shielded methylol) phenyl or 3,4- bis- (methylol) phenyl；Single (aminomethyl) or Two (aminomethyl) phenyl or (shielded aminomethyl) phenyl, such as 2,3 or 4- (aminomethyl) phenyl or 2,4- (shielded ammonia first Base) phenyl；Or single (N- (methane sulfonylamino)) or two (N- (methane sulfonylamino)) phenyl, such as 2,3 or 4- (N- (methylsulfonyls Base amino)) phenyl.In addition, term " substituted-phenyl " is also represented by the different disubstituted phenyl group of wherein substituent, such as 3- first The chloro- 4- hydroxyphenyls of base -4- hydroxyphenyls, 3-, 2- methoxyl group -4- bromophenyls, 4- ethyl -2- hydroxyphenyls, 3- hydroxyl -4- nitrobenzene Base, 2- hydroxyl -4- chlorphenyls etc..

Term " (substituted-phenyl) alkyl " refers to be connected to the above-mentioned substitution on a kind of in alkyl group defined above One kind in phenyl group.(substituted-phenyl) alkyl is somebody's turn to do to be connected on another structure division.The example bag of (substituted-phenyl) alkyl Include group such as what follows：Such as 2- phenyl -1- chloroethyls, 2- (4 '-methoxyphenyl) ethyl, 4- (2 ', 6 '-dihydroxy Phenyl) n-hexyl, 2- (5 '-cyano group -3 '-methoxyphenyl) n-pentyl, 3- (2 ', 6 ' -3,5-dimethylphenyl) n-propyl, 4- be chloro- 3- aminobenzyls, 6- (4 '-methoxyphenyl) -3- carboxyls (n-hexyl), 5- (4 '-aminomethyl phenyl) -3- (aminomethyl) positive penta Base, 5- phenyl -3- oxos-just amyl- 1- bases, (4- hydroxyl naphthalene -2- bases) methyl etc..

Term " virtue " or " aryl " refer to single, double or three rings the group containing fragrant homoatomic ring (i.e. hydrocarbon) form, preferably For 6 to 12 yuan, such as phenyl, naphthyl and xenyl.Unless otherwise indicated, otherwise aromatic yl group is optionally selected from by one or more Following substituent group：Alkyl (optionally substituted alkyl), alkenyl (optionally substituted), aryl (optionally substituted), heterocycle (are appointed Choose generation), halogen, hydroxyl, alkoxy (optionally substituted), aryloxy group (optionally substituted), alkanoyl (optionally substituted), Aroyl (optionally substituted), Arrcostab (optionally substituted), aryl ester (optionally substituted), cyano group, nitro, amino, substitution Amino, acylamino-, carbamate, lactams, urea, urethane, sulfonyl etc..Optionally, adjacent substituents and its connection Atom 3 to 7 yuan of rings of formation.

Term " heteroaryl " refers to there is 1 to 4 heteroatomic optionally substituted five-membered ring or hexatomic ring, and hetero atom is for example For oxygen, sulphur and/or nitrogen-atoms (individually, or with other nitrogen, sulphur or oxygen annular atom being connected).In addition, above-mentioned optionally substituted Five yuan or hexatomic ring optionally with fragrance five yuan or hexa-atomic member ring systems condense.For example, the ring optionally with such as benzene, pyrrole Five yuan of the fragrance of pyridine or triazole system etc or the fusion of hexa-atomic member ring systems.

Following member ring systems are the example of signified heterocycle (substituted or unsubstituted) functional group of term " heteroaryl "：Thiophene Base, furyl, pyrrole radicals, imidazole radicals, pyrazolyl, oxazolyl, isoxazolyls, thiazolyl, isothiazolyl, triazolyl, thiadiazoles Ji, oxadiazolyls, tetrazole radical, thiatriazole Ji, oxatriazoles base, pyridine radicals, pyrimidine radicals, pyrazinyl, pyridazinyl, oxazinyls, triazine Base, tetrazolo thiadiazine, 1,5- [b] pyridazinyls and purine radicals and benzo derivative (such as benzoxazolyl, benzothiazolyl, Benzimidazolyl and indyl).

Unless otherwise indicated, otherwise heteroaryl groups are optionally selected from following substituent group by one or more：One to It is three halogens, trihalomethyl, amino, shielded amino, amide, monosubstituted amino, disubstituted amino, carboxyl, protected Carboxyl, carboxylate, hydroxyl, shielded hydroxyl, the salt of oh group, lower alkoxy, lower alkylthio, alkyl (optionally Substitution), cycloalkyl (optionally substituted), (cycloalkyl) alkyl (optionally substituted), phenyl (optionally substituted), phenylalkyl (optionally substituted phenylalkyl).The substituent of heteroaryl groups is as defined above, or in the case of trihalomethyl Can be trifluoromethyl, trichloromethyl, trisbromomethyl or three iodomethyls.Associate in the above-mentioned substituent with heteroaryl ring in use, " lower alkoxy " refers to C₁To C₄Alkoxy base, similarly, " lower alkylthio " refer to C₁To C₄Alkylthio radicals.

Term " heterocycle ", " heterocyclic type group ", " heterocyclic group " or " heterocyclic group " are related to fully saturated, part insatiable hunger And/or complete undersaturated group, it, which is included at least one carbon atoms ring, has at least one heteroatomic fragrant ring group Group (" heteroaryl ") or non-aromatic cyclic radical (such as 3 to 13 unit monocycles, 7 to 17 membered bicyclics or 10 to 20 membered tricyclic systems, It is preferred that altogether containing 3 to 10 annular atoms).Each ring comprising heteroatomic heterocyclic group can comprising 1,2,3 or 4 be selected from nitrogen The hetero atom of atom, oxygen atom and/or sulphur atom, wherein nitrogen and sulfur heteroatom are optionally oxidized, and nitrogen heteroatom can Optionally it is quaternized.Heterocyclic group can be connected in any heteroatom or carbon atom of ring or member ring systems.In the nitrogen of heterocycle The heterocyclic group being connected on atom is referred to as the heterocycle of N- connections, and the heterocyclic group being connected on the carbon atom of heterocycle is referred to as C- The heterocycle of connection.The ring of polycyclic heterocycle can be condensed by one or more be spirally connected, bridging and/or connection.

Exemplary monocyclic heterocyclic ring radical includes：Pyrrolidinyl, pyrrole radicals, pyrazolyl, expoxy propane base, pyrazolinyl, Imidazole radicals, imidazolinyl, imidazolidinyl, oxazolyl, oxazole alkyl, isoxazoline-3-yl, isoxazolyl, thiazolyl, thiadiazolyl group, Thiazolidinyl, isothiazolyl, isothiazole alkyl, furyl, tetrahydrofuran base, thienyl, oxadiazolyl, piperidinyl, piperazinyl, 2- oxopiperazinyls, 2- oxo-piperidine bases, 2- oxo-pyrrolidines base, 2- oxo azatropylidene bases (2-oxoazepinyl), azatropylidene Base, 4- piperidone bases, pyridine radicals, pyrazinyl, pyrimidine radicals, pyridazinyl, triazine radical, THP trtrahydropyranyl, tetrazole radical, triazolyl, Quinoline base, thiomorpholine base, thiomorpholine base sulfoxide, thiomorpholine base sulfone, DOX and tetrahydrochysene -1,1- dioxythiophene base etc..

Exemplary bicyclic heterocyclic group includes：Indyl, benzothiazolyl, benzoxazolyl, benzothienyl, quinine Ring group, quinolyl, tetrahydro isoquinolyl, isoquinolyl, benzimidazolyl, benzopyranyl, indolizine base, benzofuranyl, benzene And furfuran base (benzofuranly), dihydro benzo furyl, chromone base, cumarin base, benzo dioxolen base (benzodioxolyl), dihydrobenzo dioxolen base (dihydrobenzodioxolyl), Ben Bing bioxin bases (benzodioxinyl), cinnolines base, quinoxalinyl, indazolyl, pyrrolopyridinyl, furopyridyl (such as furans simultaneously [2, 3-c] pyridine radicals, furans simultaneously [3,2-b] pyridine radicals or furans simultaneously [2,3-b] pyridine radicals), dihydro-iso indolyl, dihydroquinazoline Base (such as 3,4- dihydro -4- oxo-quinazolinyls), tetrahydric quinoline group, (such as 6- azabicyclos [3.2.1] are pungent for azacycloalkyl Alkane), azaspiro alkyl (such as Isosorbide-5-Nitrae dioxa -8- azaspiros [4.5] decane), imidazopyridyl (such as imidazo [1,5-a] pyrrole Pyridine -3- bases), triazolo pyridyl (such as 1,2,4- triazols [4,3-a] pyridin-3-yl) and hexahydro imidazopyridyl (such as 1, 5,6,7,8,8a- hexahydro imidazo [1,5-a] pyridin-3-yls) etc..

Exemplary tricyclic heterocyclic groups include：Carbazyl, benzindole base (benzidolyl), phenanthroline, acridine Base, phenanthridinyl, folder xanthyl etc..

Unless otherwise indicated, otherwise heterocyclic group is optionally selected from following substituent group by one or more：Alkyl (bag Include substitution alkyl), alkenyl, oxo, aryl (including substituted aryl), heterocycle (including substituted heterocycle), carbocyclic ring (optionally substituted), Halogen, hydroxyl, alkoxy (optionally substituted), aryloxy group (optionally substituted), alkanoyl (optionally substituted), aroyl (are appointed Choose generation), Arrcostab (optionally substituted), aryl ester (optionally substituted), cyano group, nitro, acylamino-, amino, substitution ammonia Base, lactams, urea, urethane, sulfonyl etc., wherein optionally, one or more pairs of substituents formed together with the atom of connection 3 to 7 yuan of rings.

The alkyl group that term " alkanoyl " refers to be connected to carbonyl group (can be as described above optionally substituted ) (that is ,-C (O)-alkyl).Similarly, term " aroyl " refers to that the aromatic yl group for being connected to carbonyl group (can be as described above As, it is optionally substituted) (that is ,-C (O)-aryl).

Term " (monosubstituted) amino " refers to the amino with a substituent in the group being made up of following group Group：Phenyl, substituted-phenyl, alkyl (including substitution alkyl), C₁To C₄Acyl group, C₂To C₇Alkenyl (including C₂To C₇Replace alkene Base), C₂To C₇Alkynyl, C₇To C₁₆Alkylaryl (including C₇To C₁₆Replace alkylaryl) and heteroaryl groups.It is somebody's turn to do (monosubstituted) Amino can also have amido protecting group, as term " shielded (monosubstituted) amino " is covered.Term is " (double to take Generation) amino " refer to the amino group with two substituents in the group being made up of following group：Phenyl, substituted-phenyl, Alkyl, substitution alkyl, C₁To C₇Acyl group, C₂To C₇Alkenyl, C₂To C₇Alkynyl, C₇To C₁₆Alkylaryl, C₇To C₁₆Replace alkyl virtue Base and heteroaryl.Described two substituents may be the same or different.

Term " heteroaryl (alkyl) " refers to the alkane defined above replaced at an arbitrary position by heteroaryl defined above Base group.

" isostere " is not homoatomic, molecule or ion with different molecular formula, but it has similar or phase Same outer-shell electron arrangement, and also there is similar characteristic (such as pharmacological characteristics (such as pharmacokinetics and pharmacodynamics spy Property)).

" structure division " (moiety) refers to particular section or the functional group of molecule.Chemical moieties are typically embedded into point Recognizable chemical unit in son or with attaching molecules.

As used herein, term " R " and its relational language R1, R2, R3 etc. refer to substituent, as herein defined that Sample.

" sulfate radical " refers to SO₃ ^-。

" nitro " refers to-NO₂Base.

" oxa- " refers to-O- bases.

" oxo " refers to=O bases.

" sulfonyl " refers to following groups：-S(O₂)-H、-S(O₂)-(alkyl) ,-S (O₂)-(cycloalkyl) ,-S (O₂)-(ammonia Base) ,-S (O₂)-(aryl) ,-S (O₂)-(heteroaryl) and-S (O₂)-(Heterocyclylalkyl)." sulfonamido " Or " sulfonamido " refers to-S (=O) (Sulfonamidyl)₂- NRR bases, wherein each R is independently selected from by following group The group of composition：Hydrogen, alkyl, cycloalkyl, aryl, heteroaryl (being connected by the carbon on ring) and heteroalicyclyl are (by ring Carbon is connected).- S (=O)₂R group in the-NRR of-NRR bases can form 4,5,6 or 7 yuan of rings (- S together with connected nitrogen (O₂)-(Heterocyclylalkyl).It is C in some embodiments₁-C₁₀Each R in sulfonamido, wherein sulfonamido is wrapped altogether Containing 1,2,3 or 4 carbon.Sulfonamido is optionally by one or more such as herein for alkyl, cycloalkyl, virtue The described substituent substitution of base, heteroaryl difference." sulfone " refers to-S (O₂)-(alkyl) ,-S (O₂)-(aryl) ,-S (O₂)-(heteroaryl Base) or-S (O₂)-(Heterocyclylalkyl) (at this moment, sulfone group is attached on the carbon atom of Heterocyclylalkyl).Sulfonamido is optionally Replaced by one or more substituents as described in herein for alkyl, cycloalkyl, aryl, heteroaryl difference.

As used herein, " label " and its form of presentation grammatically deformed refer to detectable compound or combination Thing, it is directly or indirectly conjugated with another compound, such as conjugated with saxitoxin analog, so as to produce " mark " change Compound.Label includes detectable structure division.Detectable structure division can directly or indirectly produce detectable Signal.Label can be (such as radioisotopic tracer or fluorescent marker) that itself can be detected, or, In the case of enzyme marker, substrate compounds can be catalyzed or detectable chemical modification occurs for composition.

Compound can with provide detectable signal label it is directly or indirectly conjugated, the label is, for example, radioactivity Isotope, fluorescer, enzyme, antibody, particle such as magnetic-particle, chemical luminophor, quantum dot or specific binding molecules etc..It is excellent The label of choosing includes but is not limited to fluorescent marker, marker enzyme and radio isotope.Suitable label includes：For example, Fluorescence or chemiluminescence compound, such as fluorescein, fluorescein isothiocynate, rhodamine, tetramethylrhodamine, Yihong, algae red, perfume (or spice) Legumin, methylcoumarin, pyrene, peacock green (Malacite green), stibene, fluorescein (Lucifer Yellow), CASCADETEXASIAEDANS, EDANS, BODIPY FL, LC red 640, Cy 5, Cy 5.5, LC are red 705 and OREGON GREENTM.In the written Molecular Probes Handbook (1996) of Richard P.Haugland In describe suitable optical dye, clearly its content is incorporated herein by reference herein.

Suitable label also includes：Fluorescin or peptide (such as green fluorescent protein (GFP) and GFP variants and related Peptide)；(GFP, referring to document：Chalfie etc., Science263 (5148)：802-805 (on 2 11st, 1994)；And EGFP, Clontech company-Genbank accession number (Genbank Accession Number) U55762), blue fluorescent protein (BFP；Quantum Biotechnologies companies, address is：1801de Maisonneuve Blvd.West, 8th Floor, Montreal (Quebec) Canada H3H 1J9；Referring to document：Stauber, R.H.Biotechniques 24 (3)：462-471(1998)；3.Heim, R.and Tsien, R.Y.Curr.Biol.6：178-182 (1996)), enhanced Huang Color fluorescin (EYFP；Clontech Laboratories companies, address is 1020East Meadow Circle, Palo Alto, Calif.94303), luciferase is (referring to document：Ichiki etc., J.Immunol.150 (12)：5408-5417 And Renilla WO 92/15673 (1993)；WO 95/07463；WO 98/14605；WO 98/26277；WO 99/ 49019；United States Patent (USP) 5,292,658；United States Patent (USP) 5,418,155；United States Patent (USP) 5,683,888；United States Patent (USP) 5,741, 668；United States Patent (USP) 5,777,079；United States Patent (USP) 5,804,387；United States Patent (USP) 5,874,304；United States Patent (USP) 5,876,995； And United States Patent (USP) 5,925,558).

Suitable label also includes：Radioactively labelled substance is (such as¹²⁵I、²⁵S、³²p、¹⁸F、¹⁴C、³H etc.)；Biomarker, Including specific binding molecules (such as biotin, Streptavidin, digoxin and anti-digoxin), enzyme such as alkaline phosphate ester Enzyme, beta galactosidase are (referring to document：Nolan etc., Proc Natl Acad Sci USA 85 (8)：2603-2607(1988 April in year)) or horseradish peroxidase；And other labels.All above-mentioned documents are expressly incorporated in this by reference Wen Zhong.

Any of method in this area can be used, by detectable structure division and to or to be incorporated to saxitoxin similar It is in thing or the saxitoxin analog and detectable structure division is conjugated, including those methods described in documents below： Hunter etc., Nature, 144：945(1962)；David etc., Biochemistry, 13：1014(1974)；Pain etc., J.Immunol.Meth., 40：219(1981)；And Nygren, J.Histochem.and Cytochem., 30：407 (1982)。

Terminology used in this article " antibody " has broadest sense, and it includes：For example, polyclonal antibody, Dan Ke Grand antibody, with the specific antibody compositions of multi-epitope, single-stranded and antibody fragment.As used herein, " monoclonal resists term Body " refers to the antibody obtained from a group substantially homologous antibody, i.e. except may the change that exists of spontaneous very small amount Different, the single antibody of this group is identical.

" being specifically bound to " or to as the antibody of the epitope " there is specificity " of particular polypeptide or particular polypeptide Antibody, it is attached to the epitope of the particular polypeptide or particular polypeptide, without be substantially attached to any other polypeptide or The epitope of polypeptide.

As used herein, term " antibody fragment " and its form of presentation grammatically deformed refer to include the one of complete antibody Partial molecule, is preferably the antigen binding domain or variable region of the complete antibody.The example of antibody fragment includes Fab, Fab ', F (ab ') .sub.2 and Fv fragments；Diabodies (diabodies)；Linear antibodies are (referring to document：Zapata etc., Protein Eng.8(10)：1057-1062[1995])；Single-chain antibody molecules；And the polyspecific formed by antibody fragment Antibody.Papain digestion of antibodies, produces two identical antigen-binding fragments and (is referred to as " Fab " fragment, each has one Single antigen binding site), and " Fc " residue segment (mark of reflection crystallization difficulty or ease ability).Pepsin is produced F (ab ') with two antigen binding sites₂Fragment, and the fragment still can make antigen crosslinking.

As used herein, term " Fv " refers to the minimum antibody fragment of the identification comprising comlete antigen and binding site.Should Region is made up of the dimer of a weight chain variable district and a light chain variable district closely noncovalent associations.Exactly at this Plant in construction, three CDR of each variable region interact and limit V_H-V_LThe antigen binding site of dimer interface.It is overall next Say, six CDR assign antibody with antigen-binding specificity.Even however, single variable region (or Fv half, containing only confrontation Original has specific three CDR) it can also recognize and combine antigen, but it is lower than the affinity of entire binding site.

As used herein, term " Fab fragments " refers to the antibody containing constant region of light chain and the constant region of heavy chain first (CH1) Fragment.The difference of Fab fragments and Fab ' fragments is：Carboxyl terminal of the Fab ' fragments in heavy chain CH1 areas with the addition of several Residue (including one or more cysteines from antibody hinge region).The cysteine residues of constant region are carried into mercapto herein The Fab ' of base free radical is designated as Fab '-SH.Initially, F (ab ')₂Antibody fragment, which is prepared to a pair, has therebetween hinge half Fab ' the fragments of cystine.Other chemical coupling modes of antibody fragment are also known in the art.

" light chain " of antibody (immunoglobulin) from any vertebrate can be according to the amino acid sequence of their constant regions The one kind for arranging and being classified as in two kinds of entirely different types, described two types are referred to as κ and λ.

" scFv " or " sFv " antibody fragment includes the V of antibody_HAnd V_LArea, these regions are present in single polypeptide chain. The Fv polypeptides, which are preferably included, is located at V_HAnd V_LPeptide linker between area, the peptide linker can make sFv formation combine antigen institute The structure needed.Summary on sFv can be found in document：Rosenburg and Moore, Pluckthun in The Pharmacology of Monoclonal Antibodies, volume 113, Springer-Verlag, New York, pp.269-315(1994)。

According to the amino acid sequence of heavy chain constant region, immunoglobulin can be divided into variety classes.Mainly there are five kinds to be immunized Globulin：IgA, IgD, IgE, IgG and IgM, and some of which can also be divided into hypotype (isoform), such as IgG1, IgG2, IgG3, IgG4, IgA and IgA2.

As used herein, term " Diabodies " refers to the small antibody fragment with two antigen binding sites, The fragment includes in same polypeptide chain and is connected to light chain variable district (V_L) weight chain variable district (V_H).By on same chain Described two regions between prevent it using very short from the joint that matches, so as to force to make the region and another chain Complementary region is matched, and produces two antigen binding sites.Diabodies are described in more detail in the following documents, The document is, for example,：EP 404,097；WO 93/11161；And Hollinger etc., Proc.Natl.Acad.Sci.USA, 90：6444-6448(1993).

Saxitoxin similar compound as described herein may connect on another molecule, and another molecule can be used as mark Thing and/or saxitoxin analog can be directed to ad-hoc location, organ, tissue, cell or Cytology Lab.For example, antibody or Antibody fragment may be connected on saxitoxin analog, so that saxitoxin analog effectively is directed into specific electricity Press gated sodium channel isoform or expression Na_VCell type.As used herein, phrase " guides saxitoxin analog To ... " in " guiding " be used for show saxitoxin analog can be combined in or can be positioned at desired location, organ, tissue, The vicinity of cell or Cytology Lab.

As used herein, term " imaging " refers to the part that subject or subject are produced using imaging device and method Image, such as x-ray imaging, ct (CAT) imaging, positron emission computerized tomography (PET) imaging, magnetic Resonance image-forming (MRI), single photon emission computed tomography (SPECT) imaging etc..

For example, PET imagings include setting up the faultage image of the positron-emitting radioactive nucleic of target subject.Will radiation The medicine of property isotope labeling, i.e. radiopharmaceutical, are applied to the subject being imaged.The subject is placed in PET imagings In system, the imaging system includes detection ring and detection electronic equipment.Due to the decay of radionuclide, launch and be referred to as The positively charged photon of " positive electron ".For conventional radiopharmaceutical, such as FDG is (i.e.¹⁸F- fluorodeoxyglucoses), these Positive electron can only pass through subject several millimeters of histokinesis, afterwards with electron collision and cause to bury in oblivion mutually.Positive electron/the electricity Son buries in oblivion the gamma rays that can cause to launch that a pair of reverse energy are about 511keV.

What the scintillator component of detection ring to be detected is exactly these gamma rays.When being hit by gamma ray, Scintillation material in the component sends light, and it is detected by photodetector assembly (such as photodiode or photomultiplier). The signal of photodetector is handled with the incidence of gamma rays.When two gamma rays hit almost identical at the time of it is opposite During the scintillator of position, record meets event next time.Data sorting unit handles this and meets event to determine really to meet Event, and pick out those data for representing dead time and single gamma ray detection.Event binarization will be met and shape is integrated Into PET data frame, it can be redeveloped into the image of distribution situation of the medicine for showing radioisotope labeling in subject's body.

MRI is such a medical imaging mode, and it can not use X-ray or other ionising radiations and build in human body Portion's image.MRI sets up one strong, homogeneous static magnetic field (i.e. " main field ") using strong magnet.When the one of human body or human body When being partially disposed in main field, the nuclear spin polarization related to the hydrogen nuclei in tissue water.It means that with these spin phase The magnetic moment of pass is preferably arranged along the direction of main field, causes small net tissue to magnetize along the axle (being conveniently " Z axis "). MRI system also includes so-called gradient coil part, and it produces spatial variations magnetic field more by a small margin when a current is applied.Generally In the case of, the magnetic-field component that gradient coil design is arranged into generation along Z axis, and its amplitude is with the one along along X, Y or Z axis Position and linear change.The effect of gradient coil is to make magnetic field intensity along single axial oblique line rises and makes core therewith by a small margin Oblique line rises spin resonance frequency by a small margin.Three have the gradient coil of normal axis by producing letter in each position of body Number resonant frequency and be used for " space encoding " MR signals.Radio frequency (RF) coil be used for produce the resonant frequency of hydrogen nuclei or its Neighbouring RF energy pulses.The coil is used to increase energy in a controlled manner for nuclear spin system.Due to loose after nuclear spin Relaxation returns to Resting Energy state, therefore they release energy in the form of radiofrequency signal.MRI system detects the signal, and adopts It is with computer and algorithm known that it is combined with multiple such other signal, for rebuilding MR images.

Discuss

Embodiment disclosed in the present application provides the method and composition for being used for studying, be imaged and treat pain.This The disclosed embodiment of application, which is provided, can be used for research pain, treatment pain and saxitoxin similarization being imaged to pain The preparation side of compound (such as saxitoxin type compound, N-STX type compound and gonyatoxin type compound) Method.Particularly, embodiment disclosed in the present application provides the stone room in vivo with sodium-ion channel antagonist function effect Clam toxin similar compound.Embodiment disclosed in the present application provides and prepares the new and new of saxitoxin similar compound The method of newness.The embodiment of this method allows to carry out selective modification to structure, so that the compound of design can have Improved receptor-specific and/or pharmaco-kinetic properties.In addition, embodiment disclosed in the present application, which is provided, prepares saxitoxin The fluorescence of similar compound and/or the method for radio-labeled forms, so that saxitoxin analog (including knee ditch algae Toxin type compound) available for the position and/or source that pain is imaged and determined to pain.In addition, can be by saxitoxin class It is designed to show inhibition to specific sodium channel isoform like the embodiment of compound.

Structure A

The embodiment of the compound is included in structure A, its isomer, its dynamic isomer or these materials The pharmaceutically useful salt of any one in the pro-drug of any one or these materials.

R1 can be group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,= O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、- SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, It is halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, complete It is fluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, miscellaneous Cyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), Nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

R2 can be group such as what follows：Hydrogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo- Substituted alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) Alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, heteroaryl (alkyl), oxa-, oxo, sulfonyl, Sulfonamido, sulfone ,-OR_A,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、- SO₂R_A、-NO₂Or-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, alkyl, alkenyl, alkynes Base, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (ring Alkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulphur Acyl group, sulfonamido or sulfone.

R3 can be group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、-SO₂R_A、- N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, alkyl, Alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkanes Base, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkane Acyl group, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxygen Generation, sulfonyl, sulfonamido or sulfone.

R4 can be group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、-SO₂R_A、- N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, alkyl, Alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkanes Base, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkane Acyl group, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxygen Generation, sulfonyl, sulfonamido or sulfone.

R5 can be group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfate radical, sulfonamido, Sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、- SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AIt can independently be such as what follows Group：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, perhalogeno Substituted alkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocycle Type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl Base (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

R6 can be group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfate radical, sulfonamido, Sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、- SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AIt can independently be such as what follows Group：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, perhalogeno Substituted alkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocycle Type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl Base (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

R7 can be group such as what follows：Hydrogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo- Substituted alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, (substituted-phenyl) alkyl, virtue Base, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, Asia Sulfonamido, sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、- CN、-SO₂R_A、-NO₂Or-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, alkyl, alkene Base, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkanes Base, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkane Acyl group, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxygen Generation, sulfonyl, sulfonamido or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl, alkoxy, cyano group, heteroaryl, (list Substitution) amino, shielded (monosubstituted) amino, (disubstituted) amino ,-OR_A,=O ,-OC (=O) R_A,-OC (=O) OR_A、- OC (=O) N (R_A)₂、-CN、-_SCN、-SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂Or-NHC (O) R_A, wherein each R_ACan be independent Ground is group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, saturated dihalide Base, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkane Base, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) ammonia Base, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

R2 can be hydrogen.R3 can be hydrogen or n-propyl.R4 can be hydrogen or n-propyl.

R5 can be group such as what follows：Hydrogen, sulfate radical ,-OR_A,=O ,-OC (=O) R_A,-OC (=O) OR_A、-OC (=O) N (R_A)₂, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alcoxyl Base, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkane Base, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) Amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, Asia Sulfonamido or sulfone.

R6 can be group such as what follows：Hydrogen, sulfate radical ,-OR_A,=O ,-OC (=O) R_A,-OC (=O) OR_A、-OC (=O) N (R_A)₂, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alcoxyl Base, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkane Base, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) Amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, Asia Sulfonamido or sulfone.

R7 can be group such as what follows：Hydrogen, alkoxy, alkanoyl ,-(CH₂)₂C (=O) R_A、-(CH₂)₂C (=O) N(R_A)₂, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyanogen Base, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, take For phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, Shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfenyl ammonia Base or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl ,-OC (=O) NHR_A,-OC (=O) N (R_A)₂, wherein each R_AGroup such as what follows can independently be：Hydrogen, methyl, isopropyl, myristyl or following group In any one：

R2 is hydrogen.R3 can be group such as what follows：Hydrogen or propyl group.R4 can be group such as what follows：Hydrogen Or propyl group.R5 can be group such as what follows：Hydrogen, sulfate radical ,-OC (=O) R_A, wherein R_AFor n-propyl or phenyl.R6 can For group such as what follows：Hydrogen, sulfate radical ,-OC (=O) R_A, wherein R_AFor n-propyl or phenyl.R7 can be such as following Described group：Hydrogen or-(CH₂)₂C (=O) N (R_A).Each Rs of the R2 into R7_ABase such as what follows can independently be Group：Hydrogen, hydroxyl or alkoxy.

Structure B

The embodiment of the compound is included in structure B, its isomer, its dynamic isomer or these materials The pharmaceutically useful salt of any one in the pro-drug of any one or these materials.Structure B is general for GTX-3 Structure.Referring to Fig. 1.

R1 can be group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,= O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、- SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, It is halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, complete It is fluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, miscellaneous Cyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), Nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl, alkoxy, cyano group, heteroaryl, (list Substitution) amino, shielded (monosubstituted) amino, (disubstituted) amino ,-OR_A,=O ,-OC (=O) R_A,-OC (=O) OR_A、- OC (=O) N (R_A)₂、-CN、-SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂Or-NHC (O) R_A, wherein each R_AIt can independently be Group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, three Haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, virtue It is base, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (double Substitution) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl ,-OC (=O) NHR_A,-OC (=O) N (R_A), wherein each R_AGroup such as what follows can independently be：Hydrogen, methyl or myristyl.

Structure C

The embodiment of the compound includes structure C (Fig. 1), its isomer, its dynamic isomer or these things The pharmaceutically useful salt of any one in the pro-drug of any one in matter or these materials.Structure C is GTX-2 General structure.Referring to Fig. 1.

R1 can be following group：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, saturated dihalide Base, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkane Base, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) ammonia Base, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、-SO₂R_A、- NO₂、-N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, halogen, Alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluor alkane Base, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocycle shape Group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, Oxa-, oxo, sulfonyl, sulfonamido or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl, alkoxy, cyano group, heteroaryl, (list Substitution) amino, shielded (monosubstituted) amino, (disubstituted) amino ,-OR_A,=O ,-OC (=O) R_A,-OC (=O) OR_A、- OC (=O) N (R_A)₂、-CN、-SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂Or-NHC (O) R_A, wherein each R_AIt can independently be Group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, three Haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, virtue It is base, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (double Substitution) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl ,-OC (=O) NHR_A,-OC (=O) N (R_A), wherein each R_AGroup such as what follows can independently be：Hydrogen, methyl or myristyl.

Structure D

The embodiment of the compound includes structure D (Fig. 1), its isomer, its dynamic isomer or these things The pharmaceutically useful salt of any one in the pro-drug of any one in matter or these materials.Structure D is the one of saxitoxin As structure.Referring to Fig. 1.

R1 can be group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,= O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、- SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AOr-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, It is halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, complete It is fluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, miscellaneous Cyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), Nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl, alkoxy, cyano group, heteroaryl, (list Substitution) amino, shielded (monosubstituted) amino, (disubstituted) amino ,-OR_A,=O ,-OC (=O) R_A,-OC (=O) OR_A、- OC (=O) N (R_A)₂、-CN、-SCN、-SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂Or-NHC (O) R_A, wherein each R_ACan be independent Ground is group such as what follows：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, saturated dihalide Base, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkane Base, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) ammonia Base, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido or sulfone.

In one embodiment, R1 can be group such as what follows：Hydroxyl ,-OC (=O) NHR_A,-OC (=O) N (R_A), wherein each R_AGroup such as what follows can independently be：Hydrogen, methyl, isopropyl, myristyl or following group In any one：

In structure A, structure B, structure C and structure D in the embodiment of any one, group (such as R1 to R7 groups, As each structure is applicable) be not selected into obtain with saxitoxin, N-STX, gonyatoxin or The compound of atelopidtoxin AB structures.In other words, composition, pharmaceutical composition, treatment method etc. may include structure A, structure B, One or more of structure C or structure D, precondition is：R group, which is not selected into, obtains saxitoxin, new Saxidomus Any one in toxin, gonyatoxin or atelopidtoxin AB.In one embodiment, to structure A, structure B, structure C Selected with the R group of each in structure D, to exclude corresponding to saxitoxin, N-STX, knee ditch algae poison The structure of element or atelopidtoxin AB.It is shown in Figure 1 on these structures, do not select these structures disclosed herein to obtain Noval chemical compound.

The method treated to subject can be also used described in the discussion to structure A-D and this paper relevant discussions Compound each embodiment pro-drug.Exemplary pro-drug can be (e.g., following by the enzyme activation in liver Ring -1,3- propane base ester of substituent group, the group is to promote the group that oxidative cleavage reacts by CYP3A etc.).These are repaiied Decorations can make composition disclosed in the present application inactivate before liver is arrived at or activity reduce (referring to document：Current Opinion The No 2,109-117 of in Investigational Drugs 2006Vol 7；J.Med.Chem.2008,51,2328-2345； And Nucleosides, Nucleotides, and Nucleic Acids, 24 (5-7)：375-381, (2005), passes through reference The content accordingly discussed is incorporated into herein by mode).

Synthetic method

In general, the embodiment that the present invention provides compound (is included in the discussion to structure A-D in Fig. 1 and sheet Apply for each chemical embodiments described in relevant discussion) can be according to organic synthesis technology well known by persons skilled in the art And/or the synthetic schemes provided according to the present invention is carried out.Specifically, provided herein is embodiment provide prepare the present invention The details of disclosed compound and guidance.

As needed, the compound described in the commercially available chemicals and/or Chemistry Literature proceeds by title The synthesis of compound." commercially available chemicals " can be obtained by following standard available channel, including：Acros Organics (being located at city of Pennsylvania, America Pittsburgh), Aldrich Chemical (are located at Wisconsin, USA close Er Woji cities, including Sigma Chemical and Fluka), Apin Chemical Co., Ltd.s (be located at Britain Milton Park), Avocado Research (opening Xia Jun positioned at Britain's indigo plant), BDH companies (being located at Toronto city), Bionet (are located at English State Cornwall prefecture), Chemservice Co., Ltds (be located at Pennsylvania, America west chester city), Crescent chemistry Company's (being located at New York, United States Hauppauge cities), Eastman organic chemical reagents, Eastman Kodak Companies (are located at U.S. New York Rochester city of state), Fisher scientific ＆ technical corporation (be located at city of Pennsylvania, America Pittsburgh), Fisons chemistry it is public Department's (being located at Leicestershire,England), Frontier Scientific (being located at Utah, USA Logan cities), ICN biologics have Limit company (be located at U.S. Jia Lifoniya states Coase tower Metz city), Key Organics (being located at Cornwall, UK), Lancaster Synthesis (being located at New Hampshire Windham cities), Maybridge Chemical Co., Ltd.s (are located at Cornwall, UK), Parish chemical companies (be located at Utah, USA Ao Lemu cities), Pfaltz＆Bauer Co., Ltds (position In Connecticut, USA Waterbury city), Polyorganix (be located at tennessee,USA Houston city), Pierce chemistry Company's (be located at Illinois, USA Rockford city), Riedel de Haen AG (being located at Hanoverian, Germany), Spectrum Quality Product companies (being located at New Jersey Xin Binshi Brunswicks city), TCI America (are located at U.S. Portland, Oregon city of state), Trans World chemical companies (be located at Maryland, USA Rockville city) and Wako Chemicals USA companies (are located at Virginia, The United States state Ritchie Meng Shi).

In addition, method known to those skilled in the art can be determined by various reference books and database.Retouch in detail The synthesis available for the reactant for preparing the compounds of this invention is stated or those there is provided the reference papers for describing to prepare is suitable Reference book and paper include：For example, " Synthetic Organic Chemistry ", John Wiley＆Sons, Inc., New York；S.R.Sandler etc., " Organic Functional Group Preparations ", the second edition, Academic Press, New York, 1983；H.O.House, " Modern Synthetic Reactions ", the second edition, W.A.Benjamin, Inc.Menlo Park, Calif.1972；T.L.Gilchrist, " Heterocyclic Chemistry ", the second edition, John Wiley＆Sons, New York, 1992；J.March, " Advanced Organic Chemistry：Reactions, Mechanisms and Structure ", fourth edition, Wiley-Interscience, New York, 1992.The ginseng prepared available for the synthesis for the reactant for preparing the compounds of this invention or there is provided description is described in detail Examining other suitable reference books and paper of paper includes：For example, Fuhrhop, J. and Penzlin G. " Organic Synthesis：Concepts, Methods, Starting Materials ", Second, Revised and Enlarged Edition(1994)John Wiley&Sons ISBN：3-527-29074-5；Hoffman, R.V. " Organic Chemistry, An Intermediate Text " (1996) Oxford University Press, ISBN 0-19- 509618-5；Larock, R.C. " Comprehensive Organic Transformations：A Guide to Functional Group Preparations ", the second edition (1999) Wiley-VCH, ISBN：0-471-19031-4； March, J. " Advanced Organic Chemistry：Reactions, Mechanisms, and Structure " fourth editions (1992) John Wiley＆Sons, ISBN：0-471-60180-2；Otera, J. (editor) " Modern Carbonyl The Chemistry second editions (2000) Wiley-VCH, ISBN：3-527-29871；Patai, S. " Patai ' s 1992Guide to the Chemistry of Functional Groups”(1992)Interscience ISBN：0-471-93022-9； Solomons, T.W.G. " Organic Chemistry " the 7th edition (2000) John Wiley＆Sons, ISBN：0-471- 19095-0；Stowell, J.C., " Intermediate the Organic Chemistry " second editions (1993) Wiley- Interscience, ISBN：0-471-57456-2；“Industrial Organic Chemicals：Starting Materials and Intermediates：An Ullmann ' s Encyclopedia " (1999) John Wiley＆Sons, ISBN：3-527-29645-X, in 8volumes；" Organic Reactions " (1942-2000) John Wiley＆Sons, More than 55 volumes；And " Chemistry of Functional Groups " John Wiley＆Sons, 73 volumes.Unless special It is not indicated as being opposite meaning, otherwise reaction described herein is in atmospheric pressure, generally in -78 DEG C to 200 DEG C of temperature model Enclose interior progress.In addition, unless specifically stated otherwise, otherwise reaction time and condition are tended to approximately, such as reaction is in about air Carried out about 1 to about 24 hour within the temperature range of the condition of pressure and about -78 DEG C to about 110 DEG C；Remaining reaction overnight is averaged Duration is about 16 hours.

Unless specifically identified as opposite meaning, otherwise solvent used in herein described reaction is inert organic solvents. Unless specifically identified as opposite meaning, otherwise for every gram of medicament limited, 1cc (or mL) solvent constitutes volume and worked as Amount.

It is possible if desired to pass through any suitable isolated or purified process (such as filtering, extraction, crystallization, column chromatography Method, thin-layered chromatography or thick layer chromatography, or these methods combination), to implement chemical substance described herein and centre The separation and purifying of body.Embodiment can be found in illustrating for appropriate separated or separation process.But other can also be used Of equal value separates or separation method.

It should be appreciated that chemical synthesis is probably asymmetric.In case of need, if compound disclosed in the present application It is middle to there is (R) and (S) isomers, then it can be split by method known to those skilled in the art, such as by with lower section Method：Form the salt or complex compound for the non-corresponding isomery that can be separated and (such as be separated by crystallization)；Formation, which can be separated, (such as to be passed through Crystallization, gas-liquid or liquid chromatogram and separate) diastereoisomeric derivative；Make a kind of enantiomer and enantiomer specific reagent Generation selective reaction (such as enzymatic oxidation or reduction), isolates modification or unmodified enantiomer afterwards；Or in chiral ring Solution-air or liquid chromatogram process are carried out in border, the chiral environment is for example, in chiral support (as being combined with chiral ligand Silica) on, or there is chiral solvent.Alternative is can to use optically active reagent, substrate, catalyst Or solvent synthesizes specific enantiomer by asymmetric synthesis, or a kind of enantiomer is converted into separately by asymmetric conversion A kind of enantiomer.

The embodiment of herein described compound optionally contacts with pharmaceutically useful acid and forms corresponding sour addition Salt.

Many optionally substituted initial compounds and other reactants are commercially available, for example, be available from Aldrich Chemical company (is located at Milwaukee, WI, USA city)；Or those skilled in the art can be using the synthesis side generally used Method is easily prepared.

Disclosure can be synthesized by the appropriate combination of synthetic method known in the art and disclosure method Compound.Following discussion is some in the available a variety of methods of compound disclosed in the present application in order to illustrate to prepare, and It is not intended to be limited to the scope for preparing adoptable reaction or reaction sequence in compound disclosed in the present application.

These embodiments describe structure A-D synthesis, and synthesizing radioactive mark saxitoxin and knee ditch algae Toxin derivant and fluorescence saxitoxin and gonyatoxin derivative.

Application method

The application also discloses that the purposes of saxitoxin analog, including：

Use of the saxitoxin analog in the medicine for being used for being treated to the subject for needing to receive treatment is prepared On the way, the medicine include saxitoxin analog described herein, including its isomer, its dynamic isomer or The pharmaceutically useful salt of any one in the pro-drug of any one in these materials or these materials.The saxitoxin Amount of the analog in the medicine is preferably effective dose.

Saxitoxin analog is in the muscle for preparing the neuronal activity for being used for reducing subject or causing subject Purposes in the medicine loosened, the medicine is comprising saxitoxin analog described herein, its isomer, its is mutual The pharmaceutically useful salt of any one in the pro-drug or these materials of any one in tautomeric or these materials.It is described Amount of the saxitoxin analog in the medicine be preferably can effectively reduce subject neuronal activity or cause by The amount of the loosening all muscles of examination person.

Saxitoxin analog is in the muscle for preparing the neuronal activity for being used for reducing subject or causing subject Purposes in the medicine loosened, the medicine is comprising saxitoxin analog described herein, its isomer, its is mutual The pharmaceutically useful salt of any one in the pro-drug or these materials of any one in tautomeric or these materials, wherein The subject suffers from the enhanced disease of voltage-gated sodium channel.In embodiments, the enhanced disease of voltage-gated sodium channel Selected from the group being made up of following disease：Acute Pain, anal fissure, arthritis, backache, chronic ache, dentistry pain, fibromyalgia, Neuropathic pain, postoperative pain, banding blister after arthralgia, antimigraine, cervical pain, neuropathic pain, labor pain, herpes zoster It is rash, tension headache or trigeminal neuralgia pain, blepharospasm, cancer, arrhythmia cordis, epilepsy, focal dystonia, many Sweat disease, muscle cramp and atony of bladder.

Saxitoxin analog is being prepared for needing in the medicine that the subject for receiving pain therapy is treated Purposes, the medicine includes saxitoxin analog described herein, including its isomer, its tautomerism The pharmaceutically useful salt of any one in the pro-drug or these materials of any one in body or these materials.The Saxidomus Amount of the toxin analog in the medicine is preferably effective dose.

Saxitoxin analog is being prepared for needing in the medicine that the subject for receiving pain therapy is treated Purposes, wherein the pain is selected from：Acute Pain, anal fissure pain, arthritis ache, backache, blepharospasm pain, cancer pain Bitterly, chronic ache, dentistry pain, fibromyalgia, arthralgia, antimigraine, cervical pain, splanchnodynia, neuropathic pain, production Bitterly, neuropathic pain after herpes zoster, postoperative pain, sympathetic nerve maintenance pain, herpes zoster pain, tension headache, Trigeminal neuralgia pain, myositis pain, musculoskeletal pain；Pain in the back, sprain and pull pain；With for example non-ulcer of functional intestines problem Property indigestion, the NCCP pain related to irritable bowel syndrome；The pain related to myocardial ischemia；Toothache； And pain caused by dysmenorrhoea.

Saxitoxin analog is preparing the purposes in being used to diagnose the composition of subject's illnesses, the purposes bag Include and prepare a kind of composition, the composition include saxitoxin analog described herein, including its isomer, The pharmaceutically useful salt of any one in the pro-drug or these materials of any one in its dynamic isomer or these materials. In embodiments, the consumption of the saxitoxin similar compound is can be effectively enhanced by voltage-gated sodium channel Disease is positioned at the amount of specific region in subject's body.

Purposes of the saxitoxin analog in the composition for being used for being imaged to subject is prepared, the purposes includes Prepare a kind of composition, the composition includes saxitoxin analog described herein, including its isomer, its The pharmaceutically useful salt of any one in the pro-drug or these materials of any one in dynamic isomer or these materials. In embodiment, the consumption of the saxitoxin similar compound is that describedization can be effectively detected in imaging process The amount of position of the compound in subject's body.

Purposes of the saxitoxin analog in preparing for treating the medicine of wrinkle, the medicine includes the application institute The precursor of any one in the saxitoxin analog stated, including its isomer, its dynamic isomer or these materials The pharmaceutically useful salt of any one in medicine or these materials.In embodiments, the saxitoxin similar compound Measure that the amount of wrinkle can be effectively mitigated or eliminated.

The preparation of medicine and method of administration

Embodiment disclosed in the present application provides pharmaceutical composition, and the pharmaceutical composition, which is included in, begs for structure A-D By and the application relevant discussion described in chemical embodiments in one or more, and with or without pharmaceutically acceptable Excipient, diluent, carrier and/or adjuvant.In some embodiments, the pharmaceutical composition comprising the compound is formulated Into substantially free of excipient.In other embodiments, can be pharmaceutically useful by the compound or composition and one or more Auxiliary substance prepares patent medicine together.

In one embodiment, the compound can be combined with other medicaments and is combined so as to prepare disclosed in present application Thing, and said composition may include one or more pharmaceutically useful excipient, diluent, carrier and/or adjuvant.

Diversified pharmaceutically useful excipient is known in the art.Describe in detail in many publications can Medicinal excipient, including：For example, A.Gennaro (2000) " Remington：The Science and Practice of Pharmacy ", the 20th edition, Lippincott, Williams ,s ＆Wilkins；Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) H.C.Ansel etc. writes, the 7th edition, Lippincott, Williams ,s ＆Wilkins； And Handbook of Pharmaceutical Excipients (2000) A.H.Kibbe etc. writes, the 3rd edition, Amer.Pharmaceutical Assoc。

Pharmaceutically useful excipient (such as medium, adjuvant, carrier or diluent) is the public is readily available.In addition, can medicine (such as pH is adjusted and buffer, tonicity contributor (tonicity adjusting agent), stabilizer, profit auxiliary substance Humectant etc.) also it is what the public was readily available.

In an embodiment disclosed in the present application, by the compound or composition by the way that desired effects can be reached (such as Treatment pain, mitigate pain etc.) any method and be applied to main body.Therefore, compound or composition can be incorporated to for controlling In the diversified formulation for treating administration.For example can be by the compound or composition and suitable pharmaceutically useful carrier or diluent Admix and be configured to pharmaceutical composition, can also be formulated as the preparation of solid, semisolid, liquid or gas form, such as tablet, capsule Agent, pulvis, granule, paste, solution, suppository, injection, inhalant and aerosol.

In embodiments, compound or composition can be formulated as being suitable to local administration, such as liquid, gel, ointment, breast Cream, ointment, ointment, oil, paste, pulvis or the formulation of other suitable local administrations.The compound and composition can be configured to Suitable for directly applying to skin or tissue or the external or internal surface of patient or a part, or it can be configured to be suitable to It is integrated into dressing, bandage, plaster, adhesive plaster, nail (staple), conduit, pin, supply formula induction system (depot delivery System in), or be configured to be suitable to be integrated into available for by compound or composition with disclosure feature to In other devices or equipment of medicine.The part of compound disclosed in the present application or the formulation of cutaneous penetration include：Ointment, paste, Emulsifiable paste, lotion, gel, pulvis, solution, spray, inhalant or plaster.Aseptically by active component with it is pharmaceutically useful Carrier and any required preservative or the buffer mixing that may be needed.Eye-drops preparations, auristilla and eye drops are also contemplated for In the range of disclosure.In addition, disclosure, which is additionally contemplates that, uses transdermal plaster, it has extra beneficial effect Really, i.e., the controlled delivery there is provided compound in vivo.The formulation is by the way that compound to be dissolved or dispersed in appropriate medium In prepare.Sorbefacient can also be used for improving the amount through the compound of skin.Can through-rate control film or will change Compound, which is dispersed in polymer substrate or gel, carrys out speed control.

In pharmaceutical dosage form, the compound or composition can be administered in the form of its pharmaceutically useful salt, or can be independent Suitably it is combined and is applied in combination using title active agents or by it with other pharmaceutically active compounds.Following methods and Excipient is merely illustrative to be used, and is in no way intended to limit the application.

For oral formulations, the compound or composition can be used alone or be combined with suitable additive, to make Standby tablet, pulvis, granule or capsule, for example, can be combined with conventional additives, conventional additives are, for example, lactose, sweet dew Sugar alcohol, cornstarch or farina；Can be combined with adhesive, adhesive be, for example, avicel cellulose, cellulose derivative, Arabic gum, cornstarch or gelatin；It can be combined with disintegrant, disintegrant is, for example, cornstarch, farina or carboxylic first Base sodium cellulosate；It can be combined with lubricant, lubricant is, for example, talcum or magnesium stearate；If desired, can with diluent, Buffer, wetting agent, preservative and flavor enhancement combination.

Can be by the way that the compound or composition be dissolved, suspend or emulsified in aqueous or non-aqueous solvent, and by chemical combination Thing or composition are configured to ejection preparation, the solvent be, for example, vegetable oil or other similar oil, synthctic fat acid glyceride, The ester of higher fatty acids or propane diols；If desired, can be combined with conventional additives, conventional additives be, for example, solubilizer, etc. Penetration enhancer, suspending agent, emulsifying agent, stabilizer and preservative.

Compound or composition can be used in the aerosol dosage forms of administration by inhalation.Can be by compound or composition It is incorporated in the propellant that can be pressurizeed, propellant is, for example, dicholorodifluoromethane, propane, nitrogen etc..

Furthermore it is possible to by the way that compound or composition are mixed with various matrix (such as emulsified bases or water-soluble matrix) Close and suppository is made.Compound or composition can be administered by suppository per rectum.Suppository may include excipient, such as cocoa butter, card Ripple wax (carbowax) and polyethylene glycol, it melts under body temperature, but solidifies at room temperature.

Unit dosage forms orally or rectally, such as syrup, elixir and supensoid agent are can provide for, wherein each dosage list Position (such as one, a soupspoon, an a piece of or suppository) includes the composition of predetermined volume, and said composition contains this Shen Please disclosed one or more compounds.Similarly, the unit dosage forms for injection or intravenous administration can be included in combination Compound in thing, said composition is the solution in sterilized water, physiological saline or other pharmaceutically useful carriers.

Compound or composition can be configured to the composition of injectable according to disclosure.Under normal circumstances, may be used Injectable composition is prepared as liquid solution or suspension；Also solid form can be prepared into, wherein the solid form is suitable in note It is dissolved or is suspended in liquid-carrier before penetrating.Said preparation can also be emulsified or by active component (chemical combination according to disclosure Thing) it is encapsulated in liposome vectors.

In one embodiment, compound or composition are formulated as being suitable to be conveyed by continuous conveying system. Herein, term " continuous conveying system " is used interchangeably with " controlled delivery system ", and defeated including continuous (such as controlled) Device (such as pump) and conduit, the combination of injection device are sent, diversified this kind of induction system is known in the art.

Machinery or electromechanical infusion pump can also be suitably applied disclosure.The example of this kind equipment includes：For example, beautiful State patent No.4,692,147,4,360,019,4,487,603,4,360,019,4,725,852,5,820,589,5,643, 207th, those described in 6,198,966 etc..Generally, the conveying of compound can use a variety of pumping systems refilled Any one of complete.Pump provides consistent time controlled release.In some embodiments, the compound be mounted in medicine not Liquid dosage form in permeable reservoir, and individual is conveyed in a continuous manner.

In one embodiment, delivery system is at least part implantable device.It can use known in the art Method and apparatus the implantable device is implanted into any appropriate implantation site.Implantation site is introduced in subject's body With the site for placing doser.Implantation site includes but need not be confined to：It is (subdermal) under skin, subcutaneous (subcutaneous), intramuscular or other interior suitable sites of subject's body.Use and be subcutaneously implanted in some embodiments Site, is due to so to be easy to be implanted into and remove doser.

The drug release device for being suitable for using in disclosure can be based on any one of multiple modes of operation.Example Such as, drug release device can be based on diffusion system, contracurrent system or erodable system (such as corrosion type system (erosion-based system)).For example, drug release device can be electrochemical pump, osmotic pumps, electroosmotic pump, steam press pump or bursting property of infiltration Matrix (osmotic bursting matrix) is (for example, in this case, medicine is introduced into polymer, along with through medicine The degraded of the polymeric material (such as polymeric material biodegradable, through medicine dipping) of thing dipping, the polymer makes Obtain pharmaceutical preparation to be released).In other embodiments, drug release device is based on electrodiffusion system, electrolysis pump, effervescence type Pump, piezoelectric pump, hydrolysis system etc..

Drug release device based on machinery or electromechanical delivery pump can also be suitable in disclosure.This kind of device Example be included in those described in following patent document：For example, United States Patent (USP) No.4,692,147；4,360,019；4, 487,603；4,360,019；4,725,852 etc..Generally, title treatment method can using it is a variety of refill it is non-mutually Any one of formula pumping system is changed to complete.Generally preferable pump and other contracurrent systems, this is due to that they would generally be produced More consistent time controlled release.Osmotic pumps are used in some embodiments, and this is due to that it has more consistent controlled release and phase concurrently To the advantage of less size these two aspects (referring to patent document：For example, PCT Publication application No.WO 97/27840, the U.S. is special Sharp No.5,985,305 and 5,728,396).The exemplary osmotic drive type device being suitable in disclosure include but It need not be confined to those described in following patent：United States Patent (USP) No.3,760,984,3,845,770,3,916,899,3, 923,426、3,987,790、3,995,631、3,916,899、4,016,880、4,036,228、4,111,202、4,111, 203、4,203,440、4,203,442、4,210,139、4,327,725、4,627,850、4,865,845、5,057,318、5, 059,423rd, 5,112,614,5,137,727,5,234,692,5,234,693,5,728,396 etc..

In some embodiments, delivery device is implantable device.Can using methods known in the art and Delivery device is implanted into any appropriate implantation site by device.As described herein, implantation site is drawn in subject's body Enter and place the site of doser.Implantation site includes but need not be confined under skin, in subcutaneous, intramuscular or subject's body Other suitable sites.

In some embodiments, compound or composition are conveyed using implantable drug induction system, for example with It is programmable to be administered systemically.Exemplary programmable implantable system includes implantable charge pump.Exemplary is implantable Formula charge pump or can device in connection be described in following patent：Such as United States Patent (USP) No.4,350,155,5,443, 450th, 5,814,019,5,976,109,6,017,328,6,171,276,6,241,704,6,464,687,6,475,180 and 6, 512,954.Other are suitably adapted for example devices disclosed in the present application for Synchromed charge pumps (Medtronic companies).

The suitable excipient of compound or composition be, for example, water, physiological saline, glucose, glycerine, ethanol etc. and It is combined.In addition, if if needing, the excipient can contain a small amount of auxiliary substance, such as wetting agent or emulsifying agent or pH are slow Electuary.The method for preparing the formulation is well known by persons skilled in the art, or is after present disclosure is considered Obviously.Referring to document：For example, Remington ' s Pharmaceutical Sciences, Mack publishing company (position In Pennsylvania, America Easton city), the 17th edition, 1985.Under any circumstance, composition to be administered or preparation are equal Compound or composition containing the amount for being enough to obtain Expected Results in the subject's body for receiving treatment.

Composition disclosed in the present application includes the composition containing sustained release or controlled release matrix.In addition, reality disclosed in the present application The scheme of applying can be combined with other using the therapy of sustained release preparation.As used herein, sustained-release matrix is to use that enzymatic or acid can be passed through Promote the matrix that hydrolysis or the material degraded by dissolving (being usually polymer) are made.Once entering internal, the matrix is just passed through Enzyme or humoral effect.It is such as liposome, polylactide (PLA), poly- it is appreciated that sustained-release matrix is selected from biocompatible materials Glycolide (glycolic acid polymer), poly (glycolide-lactide) (copolymer of lactic acid and hydroxyacetic acid), polyanhydride, poe, polypeptide, Hyaluronic acid, collagen, chondroitin sulfate, carboxylic acid, aliphatic acid, phosphatide, polysaccharide, nucleic acid, polyaminoacid, amino acid (such as benzene Alanine, tyrosine, isoleucine), polynucleotide, polyethylene propylene (polyvinyl propylene), polyvinyl pyrrole Alkanone and organosilicon.Exemplary biodegradable matrices include polylactide matrix, PGA matrix and poly (glycolide-lactide) (copolymer of lactic acid and hydroxyacetic acid) matrix.Similarly, the sustained release preparation of embodiment disclosed in the present application can aid in HIV suppression concentration is maintained for a long time.

In another embodiment, pharmaceutical composition disclosed in the present application (and combination group can be conveyed in controlled release system Compound).Should for example, can be applied using venous perfusion, implantable osmotic pumps, transdermal plaster, liposome or other mode of administration Compound.In one embodiment, can be using pump (referring to document：Sefton (1987), CRC Crit.Ref.Biomed.Eng.14：201；Buchwald etc., (1980), Surgery 88：507；Saudek etc., (1989) .N.Engl.J.Med.321：574).Polymeric material can be used in another embodiment.In yet another embodiment, controlled release System is placed near therapeutic purpose (such as liver), so as to only need to the part using whole-body dose.In another embodiment party In case, controlled release system is placed near therapeutic purpose, so as to only need to the part using whole-body dose.Other controlled release systems It is described in following summary：Langer(1990)Science 249：1527-1533.

In another embodiment, composition disclosed in the present application (and separately or together associated with composition) includes logical Cross by compound described herein be impregnated into absorbent material (be, for example, suture, bandage and gauze) formed by combine Thing, or by being coated to solid phase material surface (such as operation nail, slide fastener and conduit) come delivering composition.It is public according to the application Open, other such induction systems are readily apparent to those skilled in the art.

It is the compound of effective dose that particular compound disclosed in the present application can be configured to comprising its application target is directed to Pharmaceutical composition.Ache for example, the unit dosage forms that compound disclosed in the present application can be configured to about 1 μ g to 10mg are used to treat Bitterly.In some embodiments, compound disclosed in the present application or composition can be configured to about 1 μ g to 20 μ g or about 20 μ g To 1mg or about 1mg to 10mg or about 10mg to 100mg and about 50mg to 500mg unit dose.Specifically, comprising The example of compound can be formulated into 0.1 μ g, 0.2 μ g, 0.5 μ g, 1 μ g, 20 μ g, 50 μ g, 100 μ g, 200 μ g, 500 μ g, 1mg, 2mg, 5mg, 10mg, 20mg, 50mg, 100mg, 200mg and 500mg unit dosage forms.In one embodiment, unit dose Type is tablet；In another embodiment, unit dosage forms are capsule.Can be fast dissolving dosage form or slow release formulation by tablet formulation. In another embodiment, unit dosage forms are liquid.

The purposes of compound and pharmaceutical composition

Titled reference compound and its pharmaceutical composition especially have in terms of research, imaging, treatment pain or pain associated disorder With.Disclosed method additionally provides the ability for targeting, diagnosing and/or studying the various diseases related to pain.This bag Atypical pain syndrome is included, such as, but not limited to：Fibromyalgia, chronic fatigue syndrome, sympathetic reflex nutrition Bad and peripheral nerve injury syndrome.

The ability that pain can be imaged provides the objective indicator for these illnesss, and there is provided determination pain And/or the degree and/or the possibility of intensity of pressure.For pain, if its pain when checking subject Originate or the unconspicuous words that originate from, it is this ability that pain is imaged to be can also allow for determining in the source of pain or origin Position.

Treatment method is generally included in the way of one or multi-agent, is treated to needing the subject for receiving pain therapy to apply Any one in the structure A-D of effective dose and the embodiment of relevant discussion.Subject for suffering from pain, generally Disclosed method can be effectively treated after a period of time by a few days, a few weeks or months (as mitigated or changing Pain).

The method disclosed in the present application for embodiments further providing prophylactic treatment pain, including the subject of needs is applied With any one in the structure A-D of effective dose and the embodiment of relevant discussion.In addition, for not being resistant to other analgesics The patient of medicine or other analgesic drug products to treatment pain it is of no avail in the case of, using compound disclosed in the present application Or composition may also can be beneficial.

The compound of the present invention can be applied to subject in the way of one or multi-agent and include the medicine of the compound Compositions.In one embodiment, the compound or composition can be applied with amount as described below：Every dose of about 1 μ G to 10mg, for example, every dose of about 1 μ g to 5 μ g, about 5 μ g are to 10 μ g, about 10 μ g to 50 μ g, about 50 μ g mg to 100 μ g, about 100 μ g To 200 μ g, about 200 μ g to 400 μ g, about 400 μ g to 800 μ g, about 800 μ g to 1mg, about 1mg to 2mg, about 2mg to 3mg, about 3mg to 4mg, about 4mg to 5mg, about 5mg to 6mg, about 6mg to 7mg, about 7mg to 8mg, about 8mg to 9mg or about 9mg extremely 10mg。

In one embodiment, compound described in every dose or the amount of composition are based on depending on per weight.For example, base Depending on per weight, compound or the amount of composition can be e.g., from about 10ng/kg, about about 15ng/kg, 20ng/ in every dose Kg, about 50ng/kg, about 100ng/kg, about 200ng/kg, about 500ng/kg, about 1 μ g/kg, about 2 μ g/kg, about 5 μ g/kg, about 10 μ g/kg, about 20 μ g/kg, about 50 μ g/kg, about 100 μ g/kg, about 200 μ g/kg, about 500 μ g/kg, about about 1mg/kg, 2mg/kg About 5mg/kg.

For example in one embodiment, the compound or composition can be applied with amount as described below：About 15ng/kg is to 150 μ g/kg, e.g., from about 15ng/kg to 30ng/kg, about 30ng/kg to 60ng/kg, about 60mg/kg to 120ng/ Kg, about 120ng/kg are to 240ng/kg, about 240ng/kg to 480ng/kg, about about 480ng/kg to 700ng/kg, 700mg/kg To 1 μ g/kg, about 1 μ g/kg to 2 μ g/kg, about 2 μ g/kg to 4 μ g/kg, about 4 μ g/kg to 8 μ g/kg, about 8 μ g/kg to 15 μ g/ Kg, about 15 μ g/kg are to 20 μ g/kg, about 20 μ g/kg to 30 μ g/kg, about 30 μ g/kg to 40 μ g/kg, the complete 60 μ g/ of about 40 μ g/kg Kg, about 60 μ g/kg are to 90 μ g/kg or about 90 μ g/kg to 120mg/kg or greater than about 120 μ g/kg.

It should be readily apparent to one skilled in the art that dosage level can be with the compound or composition, symptom specifically applied The order of severity and subject change to the difference of the neurological susceptibility of side effect.Those skilled in the art can be easy according to various means Ground determines the preferred dose of given compound.

In one embodiment, using compound described in multi-agent or composition.Using the compound or composition Frequency can be in many factors such as severity of symptom any factor and change.For example, in one embodiment, The administering mode of compound or composition is：Monthly 1 time, monthly 2 times, monthly 3 times, every 1 week (qow), 1 times a week (qw), 2 times a week (biw), 3 times a week (tiw), 4 times a week, 5 times a week, 6 times a week, every 1 day (qod), once a day (qd), one Its 2 times (bid) or three times a day (tid).As described above, in one embodiment, persistently using the compound or combination Thing.

Using compound or the duration of composition, the period of compound or composition is such as applied, can be according to such as sick Any factor in many factors such as person's development and change.For example, can be using the duration of compound or composition：About 1 day to 1 week, about 2 weeks to 4 weeks, about 1 month to 2 months, about 2 months to 4 months, about 4 months to 6 months, about 6 months to 8 The moon, about 8 months to 1 year, about 1 year to 2 years or about 2 years to 4 years or longer.

Implement disclosed method to generally include using the compound of effective dose, composition or comprising the compound Pharmaceutical composition.According to selected particular compound, the dosage regimen to be followed, whether with other compounds combination, administration Time, administration tissue and the physical transport system for carrying medicine, specific dosage can be different.

Can be using any obtainable method and the approach of suitable conveying medicine, including internal and external method, Yi Jiquan Body and topical routes of administration, subject is applied to by particular compound, composition and its pharmaceutical composition.

Method of administration is included through intranasal, intramuscular, tracheal strips, subcutaneous, intracutaneous, local application, intravenous, rectum, nose, mouth Chamber and other enterals and parenteral route of administration.Method of administration can be combined as needed, or according to the effect of medicament and/or needs And adjusted.Particular compound or composition can be applied in the way of one or multi-agent.

It can incite somebody to action this using conventionally available method and the approach (including whole body or topic route) of suitable conveying conventional medicine The disclosed specific material of application is applied to main body.Generally, the method for administration including but not limited to intestines that disclosure is considered Interior, parenteral or inhalation route.

Other parenteral route of administration beyond inhalation mode include but is not limited to, local, transdermal, subcutaneous, muscle Interior, eye socket is interior, intracapsular, backbone interior, breastbone is interior and intravenous route, that is, passes through any method of administration beyond alimentary canal.Can Parenteral administration is carried out with the whole body for realizing the compound or composition or local conveying.When needing systemic conveying, medicine The administration of preparation generally includes invasive administration or systemic Absorption formula locally administration or mucosa delivery.

Also can be by enteral administration by the compound or delivery of composition to subject's body.Enteral administration approach include but It is not limited to：Oral cavity and rectum (such as using suppository) conveying.

The method that the compound or composition are administered is included but is not limited to by skin or mucous membrane：Topical application is suitable Pharmaceutical preparation, transdermal transmission, injection and Epidermal administration.Suitable transdermal transfer approach is sorbefacient method or iontophoresis Method.In embodiments, skin, mucous membrane or other body tissues is contacted with compound just can effectively apply the chemical combination Thing.In addition, carrier, reinforcing agent and other compounds can be used for the administration for accelerating or strengthening the compound.Iontophoresis is transmitted It can be realized using commercially available " plaster ", it is by penetrating the electric pulse of intact skin within a couple of days or longer time Persistently convey its product.In some embodiments, composition disclosed in the present application by it is oral, intravenous, transdermal, sublingual, Intramuscular or anal route administration.

Embodiment

Following examples are enumerated, how to be implemented and using the complete disclosure of the present invention to be provided for those skilled in the art And description, rather than the scope for its disclosure content that inventor is thought is limited, do not really want to represent that following experiment is all yet Or unique experiment implemented.Applicant has ensured the accuracy (such as amount, temperature) of numeral used as possible, but some experiments are missed Difference and deviation can hardly be avoided.Unless otherwise stated, number is number by weight, molecular weight is weight average molecular weight, temperature In degrees celsius (DEG C), and pressure is atmospheric pressure or close to atmospheric pressure to degree.Standardized abbreviations, such as bp, base-pair can be used；Kb, Kilobase；Pl, picoliters；S or sec, second；Min, minute；H or hr, hour；Aa, amino acid；Kb, kilobase；Bp, base-pair； Nt, nucleotides；I.m., intramuscular ()；I.p., intraperitoneal ()；S.c., subcutaneous ()；Etc..

Embodiment 1：

Voltage-gated sodium channels (Na_V) indispensable effect is played in bioelectric generation and for all Life process be also required.(referring to document：For example, Hille, B.Ion Channels of Excitable Membranes, the third edition, Sinauer：Sunderland, MA, 2001).Coding has been identified in mammalian cell 10 kinds of unique sodium channel isoform (Na_V1.1-1.9, Na_X) gene (referring to document：(a) Catterall, W.A.；Yu, F.H.Genome Biology 2003,4,207.(b) Catterall, W.A.；Goldin, A.L.；Waxman, S.G.Pharm.Rev.2005,57,397).The difference of biophysical properties between these protein subtypes, they film it is dense Degree and spatial distribution define the signal characteristic of neuron.(referring to document：For example, (a) Novakovic, S.D.；Eglen, R.M.；Hunter, J.C.Trends in Neurosci.2001,24,473.(b) Lai, H.C.；Jan, L.Y.； Nat.Rev.Neurosci.2006,7,548；(c) Rush, A.M.；Cummins, T.R.；Waxman, S.G.J.Physiol.2007,579,1.) it has been generally acknowledged that the Na of exception_VFunction and/or expression are related to various disease states, bag Include arrhythmia cordis, epilepsy, neuropathic pain and congenital indifference to pain disease.(referring to document：For example, (a) Keating, M T.； Sanguinetti, M.C.Cell 2001,104,569.(b) Lossin, C.；Wang, D.W.；Rhodes, T.H.；Vanoye, C.G.；George, A.L.Jr.Neuron, 2002,34,877.(c) Rogers, M.；Tang, L.；Madge, D.J.； Stevens, E.B.Semin.Cell.Dev.Biol.2006,17,571.(d) Cox, J.J.；Reimann, F.；Nicholas, A.K.；Thornton, G.；Roberts, E.；Springell, K.；Karbani, G.；Jafri, H.；Mannan, J.； Raashid, Y.；Al-Gazali, L.；Hamamy, H.；Valente, E.M.；Gorman, S.；Williams, R.；McHale, D.P.；Wood, J.N.；Gribble, F.M.；Woods, C.G.Nature 2006,444,894.) therefore, people seek to be used for Explore the structure of protein, for adjusting specific Na_VIsoform activity and for track and Na_VRegulation and control are related to expression Dynamic event chemical tools, to further understand the Pathological Physiology related to channel function.(referring to document：For example, (a) Anger, T.；Madge, D.J.；Mulla, M.；Ridall, D.J.Med.Chem.2001,44,115.(b) Priest, B.T.；Kaczorowski, G.J.PNAS 2007,104,8205.) here, we describe our exploitations to this kind of medicament, Wherein shellfish poison (+)-saxitoxin 1 (STX) (single digit of specific Nav hypotypes receive the inhibitor that rubs) is provided for molecule blueprint (Fig. 2).So far, our research has had shown that the existing available model of the reference state with STX in passway Between difference.In addition, we establish the means for the carbamate modified forms for obtaining the toxin, and confirm the knot Structure, which changes, will not substantially reduce the binding affinity of substrate-acceptor.

The fully functional valtage-gated Na of property⁺Passage is by 1 big different poly- alpha subunit (~260kDa) and 1 or 2 auxiliary β subunits (33-36kDa) composition.(referring to document：(a) Catterall, W.A.；Yu, F.H.Genome Biology 2003,4,207.(b) Catterall, W.A.；Goldin, A.L.；Waxman, S.G.Pharm.Rev.2005,57,397.) In the case of lacking protein crystallography data, small molecule pharmacology probe is macromolecular man with protein mutant and electrophysiology Race provides a large amount of existing Structure Understandings.(referring to document：For example, Choudhary, G.；Shang, L.；Li, X.F.； Dudley, S.C.Biophys.J.2002,83,912.) these data are together with related K⁺X-ray structure (the Kcsa of ion channel And MthK) make structure Na_VThe homology model of alpha subunit is possibly realized.(referring to document：For example, (a) Lipkind, G.M.； Fozzard, H.A.Biochemistry 2000,39,8161.(b) Tikhonov, D.B.；Zhorov, B.S.Biophys.J.2005,88,184.) passage collar extension (i.e. so-called I site) include ion selectivity filter, and by It is considered the acceptor site (Fig. 2) of STX and related guanidine radicals poisonous substance.(referring to document：Llewellyn, L.E.Nat.Prod.Rep.2006,23,200.) five carboxylic acid residues be arranged in rows in the duct area (D400, E755, E403, E758, D1532, Na_V1.4 numberings), studied according to rite-directed mutagenesis, the presence of which is to realize the pass that high-affinity STX is combined Key.(referring to document：(a) Terlau, H.；Heinemann, S.H.；St ü hmer, W.；Pusch, M.；Conti, F.；Imoto, K.；Numa, S.Febs Lett, 1991,293,93.(b) Heinemann, S.H.；Terlau, H.；St ü hmer, W.；Imoto, K.；Numa, S.Nature 1992,356,441.(c) Schlief, T.；R.；Imoto, K.；Heinemann, S.H.Eur.Biophys.J.1996,25,75.(d) Chiamvimonvat, N.；P é rez-Garcia, M.T.；Tomaselli, G.F.；Marban, E.J.Physiol.1996,491,51.(e) Chiamvimonvat, N.；P é rez-Garcia, M.T.； Ranjan, R.；Marban, E.；Tomaselli, G.F.Neuron 1996,16,1037.(f) Hui, L.；McIntyre, D.； French, R.J.J.Gen.Physiol.2003,122,63.) meter set up of Lipkind and Fozzard, Dudley and Zhorov Model is calculated all to assume：The 7 of STX, 8,9- guanidines point to the sour ring of tetramino (also known as DEKA rings) of composition selective filter.(ginseng See document：(a) Lipkind, G.M.；Fozzard, H.A.Biochemistry 2000,39,8161.(b) Tikhonov, D.B.；Zhorov, B.S.Biophys.J.2005,88,184.(c) Llewellyn, L.E.Nat.Prod.Rep.2006,23, 200.) C13- urethane units, C12- hydrated ketones and 1,2,3- guanidine radicals structure divisions and vestibular ring Specific contacts between the carboxylic acid residues of (outervestibule loop) are also study hotspot.

As the starting point of research, we select to first check for the primary carbamate as hydrogen bond donor to total toxin The contribution of binding affinity.Compared to STX, the STX (dc-STX) of naturally-occurring deaminizating formyl effect reduces 20- 40%.(referring to document：(a) Koehn, F.E.；Schnoes, H.K.；Kao, C.Y.Biochim.Biophys.Acra： Biomembranes 1983,734,129.(b) Strichartz, G.R.；Hall, S.；Magnani, B.；Hong, C.Y.； Kishi, Y.；Debin, J.A.Toxicon 1995,33,723.) before this is changed by dc-STX semi-synthetic modification The achievement that palace can be rolled into a ball is confined to single succinate derivative.(referring to document：(a) Schlager, J.J.；Williams, K.C.；Hornyak, M.J.；Courtney, B.C.；Smith, J.R.Medical Defense Bioscience Review, Proceedings, Baltimore, May 12-16,1996.(b) Robillot, C.；Kineavy, D.；Burnell, J.； Llewellyn, L.E.Toxicon 2009,53,460.) due to can be with de novo formation STX so that we can arbitrarily change this The element of C13 sides.Thus, prepare and have evaluated N, N- dimethyl-STX 2 blocks Na⁺The ability of electric current.In whole-cell voltage-clamp The rat skeletal muscle passage Na of heterogenous expression is directed in form_V1.4 alpha subunit (Chinese hamster ovary celI) carries out electrophysiology measurement. (referring to document：For example, Moran, O.；Picollo, A.；Conti, F.Biophys.J.2003,84,2999.) Fig. 3 shows 10ms, relative to keeping the postimpulse electric current record of depolarising of current potential -100mV its amplitude for 100mV.Concentration is increased 2 be filled into external solution, cause peak current to decline.These data are fitted to bright wrong that (Langmuir) thermoisopleth, obtained To 2 IC₅₀For 2.1 ± 0.1nM, IC of the value no better than (+)-STX that we synthesize₅₀Record value.The result seems table Bright, the effect of the primary carbamate in natural products is not as hydrogen bond donor.(on background technology, referring to document： Tikhonov, D.B.；Zhorov, B.S.Biophys.J.2005,88,184.) this discovery also provide further explore C13 The motivation of the steric environment in the protein duct near carbamyl residue.

The tactic adjustment carried out to one of disclosed (+)-STX routes before us is had been designed that, can with preparation For the C13 urethane forms (Fig. 4) of replacement.(referring to document：Fleming, J.J.；McReynolds, M.D.；Du Bois, J.J.Am.Chem.Soc.2007,129,9964.) San Huan oxazolidones 4 represent the basis of our new synthesis plans, We assume that nucleophilicity amine can the selectively opened tension force heterocycle.This structure can (our laboratories lead at present from 9 yuan of rings guanidines 3 The material is often synthesized with ＞ 5g scale) formed by only 3 steps.4 are obtained, has proven to sequentially form C13-Troc Carbonate and make ring closure be necessary；Cause almost only to produce if using other carbonylation agents (such as phosgene, N,N'-carbonyldiimidazole) Raw C4-C13 alkene.Open loop actually successfully occurs for , oxazolidones in the presence of primary amine, obtains corresponding secondary amino group formic acid Ester.Then pass through and convert twice, closed including lewis acid mediated guanidine ring and deprotection and C12 oxidations, so that complete Into the assembling of the saxitoxin of customization.

Saxitoxin C13- derivatives 8-12 have rated to rNa_V1.4 effect (as described in Figure 5).As 8 Linear N- hexyls structure division is introduced, causes the IC compared to natural products₅₀Only slightly reduce.The result and side chain N- isopropyls Measurement result (it, which shows binding affinity, significantly reduces) the formation control of structure 9.It is not only restricted to specific theory, it appears that ammonia Carbamate structure division is maintained at quite narrow, is possible to extend in the valley of extracellular space.(referring to document： Sato, C.；Ueno, Y.；Asai, K.；Takahashi, K.；Sato, Masahiko, S.；Engel, A.；Fujiyoshi, Y.Nature 2001,409,1047.) other data, i.e. 10 from voltage-clamp recording and 12 result, support this knot By.However, it is interesting that introducing carboxylic acid residues (i.e. 11) in the distal end of saxitoxin parent nucleus reduces pharmaceutical efficacy.Hexichol The toxin 12 of ketone mark is it is intended that the mono- towering neighbouring neighbour's amino acid contacts of C13 provide direct experimental evidence.According to we institute Know, the compound is first in such any photon compatibility STX conjugates.(it has been prepared for based on tetraodotoxin Photon compatibility probe, referring to document：(a) Guillory, R.J.；Rayner, M.D.；D ' Arrigo, J.S.Science 1977,196,883.(b) Uehara, S.；Uyemura, K.Neurochem.Res.1985,10,1119.(c) Yoshida, E.； Nakayama, H.；Hatanaka, Y.；Kanaoka, Y.Chem.Pharm.Bull. (Tokyo) 1990,38,982.(d) Nakayama, H.；Hatanaka, Y.；Yoshida, E.；Oka, K.；Takanohashi, M.；Amano, Y.；Kanaoka, Y.Biochem.Biophys.Res.Commun.1992,184,900.)

In order to finally show ability that de novo formation obtains STX new models, it is used for " synthesis we have developed compound such as 10 Modify (Fig. 6) afterwards ".Despite the presence of two guanidino groups, but 10 select with n-hydroxysuccinimide (NHS) benzoic ether Selecting property is coupled and produces required acid amides 13.The coupling reaction of this final step causes baroque payload (i.e. fluorescence Group, biomarker) it is connected on STX parent nucleus, the side chain elements incompatible with the chemical constitution in addition are de- for guanidine Protection or C12 oxidations are (referring to Fig. 4).So far, we are different by structure by the coupling reaction carried out on 10 Fluorophor (14,15,16 in Fig. 7), biotin labeling (17), double ethylene imine classes (18) for photoaffinity labeling, half Guang ammonia Acid reaction functional group (19) and lipophilic sex steroid (such as cholesterol (20)) are connected to STX.Due to the change containing maleimide Compound 19 seems irreversibly to suppress voltage-gated sodium channel (referring to Fig. 8), therefore compound 19 is species of particular concern.

Obtained by Enantioselective total synthesis (+)-STX make it possible to develop unique natural products non-natural it is similar Thing.New saxitoxin analog have rated using full cell, voltage clamp technology and block Na_VThe effect of function.These It was found that having shown that the C13- ammonia for being possible to that STX is redesigned with any one in diversified different structure group Base formoxyl unit.It is such to study the three-dimensional for enabling researcher to draw access opening in more detail together with biology tool Structure.It is believed that these researchs are to realize to address inquires to and Na using by the chemical tools designed_VThe related dynamic mistake of function Bigger steps necessary in the works for journey.

Material and method are unless otherwise stated, all reagents are commercially available.Reaction is in drying nitrogen environment Under in the glassware of drying carry out.By syringe or stainless steel pipe shift air-sensitive and moisture-sensitive liquid and Solution.(about 15 support) rotary evaporation concentrates organic solution at reduced pressure conditions.Before it will use, make dichloromethane, tetrahydrochysene Furans (THF) and acetonitrile (MeCN) and DMF pass through two activated alumina pillars.Distilled with calcium hydride Pyridine.N-Boc-L- serine methylesters are prepared according to Dondone method.(referring to document：Dondoni, A.；Perrone, D.Synthesis of 1,1-Dimethylmethyl (s) -4-Formyl-2,2-Dimethyl-3-Oxazolidine- Carboxylate by Oxidation of the Alcohol.Org.Syn.2004,10,64-70.) according to Quan and The Baldwin step of method two prepares N- (to methoxybenzyl) azanol.(referring to document：(a) Quan, C.；Kurth, M.Solid- Phase Synthesis of 5-Isoxazol-4-y1- [1,2,4] oxaciazoles.J.Org.Chem.2004,69, 1470-1474.(b) Baldwin, J.E.；Cah, J.K.；Kruse, L.I.Total Synthesis of Antitumor Agent AT-125, (aS, 5S)-a-Amino-3-Chloro-4,5-Dihydro-5-Isoxazolieacetic Acid.Tetrahedron 1985,41,5241-5260.) according to Fleming method prepare β-saxitoxin phenol (β- ) and saxitoxin Saxitoxinol.(referring to document：Fleming, J.J.；Du Bois, J.A synthesis of (+)- Saxitoxin.J.Am.Chem.Soc.2006,128,3926-3927.) tert-butyl group -6- is prepared according to Phanstiel method Amino cyclohexyl amino formic acid esters.(referring to document：Gardner, R.A.；Ghobrial, G.；Nasser, S.A.；Phanstiel, O., IV Synthesis and Biological Evaluation of New Acinetoferrin Homologues for Use as Iron Transport Probes in Mycobacteria.J.Med.Chem.2004,47,4933.) basis Dijkgraaf method prepares the tert-butyl group -2- aminoethylamino formic acid esters.(Dijkgraaf, I.；Rijnders, A.Y.； Soede, A.；Dechesne, A.C.；Van Esse, G.W.；Brouwer, A.J.；Corstens, F.H.；Boerman, O.C.； Rijkers, D.T.；Liskamp, R.M.Synthesis of DOTA-Conjugated Multivalent Cyclic-RGD Peptide Dendrimers via1,3-Dipolar Cycloaddition and their Biological Evaluation：Implications for Tumor Targeting and Tumor Imaging Purposes.Org.Biomolec.Chem.2007,5,935-944.) according to Bauer, it is prepared by three (trifluoroacetic acid) boron Into the 0.5M solution in trifluoroacetic acid, and in Shi Lunke (Schlenk) bottle being stored at -5 DEG C.(referring to document：For example, Pless, J.；Bauer, W.Boron Tris (Trifluoroacetate) for Removal of Protecting Groups In Peptide Chemistry.Angew.Chem., Iht.Ed.1973,12,147-148.) in the ultrapure silica gel of Silicycle Chromatography purifying is carried out to product using forced flow chromatography on (40-63 μm).In the Varian ProStar of model 320 Upper progress semipreparative high performance liquid chromatography (HPLC).In EM Science silica gel 60F₂₅₄Thin-layer chromatography is carried out on plate (250 μm) Method.The visualization of the chromatogram to realize expansion is dyed by fluorescent quenching and by using ammonium molybdate cerium (CAM) aqueous solution.

Detected respectively under 400 and 100MHz on Varian Mercury spectrometers¹H and¹³C, or in Varian Detected respectively under 500 and 125MHz on Inova spectrometers¹H and¹³C, obtains nuclear magnetic resonance (NMR) spectrogram, and by residual solvent Signal is used as internal reference.¹H NMR datas record is as follows：Chemical shift (δ, ppm), multiplicity (s, it is unimodal；D, it is bimodal；T, it is triple Peak；Q, quartet；Quint, quintet；M, multiplet；Br, broad peak), integration, coupling constant (Hz).¹³C NMR data are to change Displacement study (δ, ppm) is reported.Record is using the film of NaCl salt pieces on Thermo-Nicolet 300FT-IR spectrometers Infrared spectrum (IR), and reported with absorption frequency.It is located at the Jasco DIP- worked under Na D lines by the way that sample is loaded In the pond of 50mm on 1000 digital polarimeters, optically-active data are obtained.Vincent of the high resolution mass spec from Stanford University Coates foundation Mass Spectrum Laboratories.

BMA.3：Dimethylamino first is added in BMA.1 (155mg, 0.27mmol) ethanol solution into 5.0mL pyridines Acyl chlorides (555 μ L, 6.0mmol, 22 equivalent).Reflux condenser is equipped with for flask, and heats mixture to 90 DEG C.At such a temperature Stir after 12h, reactant mixture is cooled to room temperature, and be concentrated under reduced pressure.Pass through silica gel chromatography (96: 4 CH₂Cl₂/MeOH→ 93∶7 CH₂Cl₂/ MeOH gradient elutions) purify oily residue, product BMA.2 needed for obtaining, be white solid (65mg, 37%).TLC R_f=0.55 (92: 8CH₂Cl₂/MeOH)；¹H NMR(CD₃CN, 400MHz, 65 DEG C) δ 7.78-7.70 (m, 4H), 7.05-6.96 (m, 4H), 6.28-6.20 (m, 2H), 5.85 (br d, 1H, J=8.2Hz), 4.82-4.74 (m, 2H), 4.67- 4.63 (m, 1H), 4.20 (dd, 1H, J=11.4,3.2Hz), 4.11 (dd, 1H, J=11.4,5.5Hz), 3.92-3.87 (m, 1H), 3.88 (s, 3H), 3.86 (s, 3H), 3.49-3.36 (m, 2H), 2.89 (s, 3H), 2.84 (s, 3H), 2.69-2.61 (m, 1H), 2.20-2.10 (m, 1H) ppm.The substance B MA.2 (17mg, 0.027mmol) isolated is dissolved in the 11: 2 of 680 μ L MeCN/H₂In O, OsCl is then added thereto₃Solution (36mM, in H₂In O, 74 μ L, 3.0 μm of ol, 0.1 equivalent).Then to obtaining 570 μ L EtOAc, a Na is added in the brown solution obtained₂CO₃(28mg, 0.027mmol, 10.0 equivalent), and a mistake Potassium acid sulfate preparation (Oxone) (125mg, 0.20mmol, 7.6 equivalent).It was observed that there is gas leniently to discharge, acutely stir Obtained light yellow suspension is mixed up to 60h.1mL Na is added dropwise₂S₂O₃The reaction is quenched in saturated solution, stirs 5min, and turn In the separatory funnel for moving on to the EtOAc containing 10mL.Collected organic layer and with 10mL EtOAc aqueous layer extracteds.Merge organic layer simultaneously Washed, dried with magnesium sulfate, and be concentrated under reduced pressure with 5mL NaCl saturated aqueous solutions.Pass through silica gel chromatography (94: 6 CH₂Cl₂/ MeOH white residue) is purified, is obtained as white solid (4mg, 24%) BMA.3, and reclaim BMA.2 (10mg, 57%).Obtain The product obtained is isomeric 5/6 bicyclic 9: 1 mixtures (HPLC determinations).Can by reversed-phase HPLC (NovaPak C18, Using 30: 70 MeCN/0.1%CF₃CO₂The H aqueous solution is as eluent, and flow velocity is 4mL/min) sample is further purified.Upper BMA.3 is eluted under the conditions of stating, retention time is 10.1min.TLC R_f=0.37 (92: 8 CH₂Cl₂/MeOH)；¹H NMR (CD₃CN, 500MHz) δ 7.75 (s, 1H), 7.74-7.69 (m, 4H), 6.99-6.94 (m, 4H), 6.57 (br s, 2H), 6.38 (br d, 1H, J=8.9Hz), 4.26 (br d, 1H, J=10.9Hz), 4.02-3.99 (m, 1H), 3.91 (d, 1H, J= 3.8Hz), 3.84 (s, 6H), 3.72 (dd, 1H, J=11.1,6.8Hz), 3.68 (ddd, 1H, J=10.8,6.7,2.4Hz), 3.57-3.47 (m, 2H), 2.85 (br s, 3H), 2.73 (br s, 3H), 2.20-2.15 (m, 1H), 1.81 (dd, 1H, J= 13.4,6.6Hz) ppm；IR (film) v 3333,1686,1578,1536,1499,1259,1202,1133,1081,853cm^-1； HRMS(ES⁺)：C₂₆H₃₅N₇O₁₀S₂Calculated value is 669.1887, and actual value is 692.1783 (MNa⁺)。

BMA.4：By 0.5M B (O₂CCF₃)₃CF₃CO₂H solution (400 μ L, 30 equivalents) is added drop-wise to equipped with BMA.3 (it is put in advance in ice-water bath) in the flask of (4.5mg, 6.7 μm of ol).Be stirred vigorously the light brown mixture, at the same with 5h when Between content is to slowly warm up to 20 DEG C.After temperature stirring 14h, solution is cooled to -78 DEG C, and 1mL MeOH is added dropwise The reaction is quenched.It is solid fraction residue by solution decompression concentration, is then re-dissolved in 1mL MeOH.Concentration should again Solution.The process is repeated once.Then the material isolated is dissolved in 0.5mL H₂In O, and it is run through 20x 100mm Ion exchange resin column (Dowex 1x 8-200^-OH types), using H₂O is used as eluent.Collect and determined by pH (~7.5-8.0) The fraction containing product, and it is 2 to be acidified to 20 μ L 1.0M HCl/water solution pH.Solutions in Freeze-drying, obtains BMA.4, is white Powder (2.0mg, 75%).¹H NMR(D₂O, 500MHz) δ 4.76 (d, 1H, J=1.4Hz), 4.31 (d, 1H, J=4.0Hz), 4.26 (dd, 1H, J=11.5,9.0Hz), 4.04 (dd, 1H, J=11.5,5.5, Hz), 3.83 (ddd, 1H, J=9.0,5.4, 1.5Hz), 3.74 (ddd, 1H, J=10.2,10.2,2.1Hz), 3.66 (ddd, 1H, J=9.8,9.8,8.4Hz), 2.91 (s, 3H), 2.86 (s, 3H), 2.39 (dddd, 1H, J=14.7,10.2,10.2,4.4Hz), 2.21 (ddd, 1H, J=15.0,8.2, 1.5Hz)；HRMS(ES⁺)：C₁₂H₂₁N₇O₃Calculated value is 311.1706, and actual value is 312.1782 (MH⁺)。

BMA.5：By β-DMC-STX BMA.4 (2.0mg, 5.2 μm of ol) and powderyMolecular sieve is mixed into 500 μ L DMSO In and stir 30min.Dicyclohexylcarbodiimide (13mg, 63 μm of ol, 12.1 equivalents) and trifluoro second are added into the mixture Sour pyridiniujm (8.2mg, 42 μm of ol, 8.1 equivalents).Thick sediment is formed immediately, and the cloudy suspension for being stirred vigorously acquisition reaches 17h.Lyophilized reactant mixture, obtains solid product, is resuspended in 1mL H₂In O and filter.In addition with 1mL H₂O is to protect Demonstrate,prove the quantitative transfer of the material.The lyophilized filtrate merged, and with the reverse HPLC-purified solid product (Altima isolated C18,10 μm, 10x 250mm posts, 10: 90 MeCN/10mM C₃F₇CO₂The H aqueous solution → 25: 75 MeCN/10mM C₃F₇CO₂H water Solution gradient elutes 30min, 214nm UV detections).Flow velocity is 6mL/min, formyl-dimethylamino saxitoxin 2C₃F₇CO₂H retention time is 20.8min, isolated white hygroscopic solid (1.2mg, 60%).¹H NMR(D₂O, 500MHz) δ 4.73 (d, 1H, J=1.0Hz), 4.29 (dd, 1H, J=11.6,9.5Hz), 4.03 (dd, 1H, J=11.6, 5.3Hz), 3.84 (ddd, 1H, J=9.3,5.2,0.9Hz), 3.78 (ddd, 1H, J=9.7,9.7,1.7Hz), 3.55 (ddd, 1H, J=9.7,9.7,8.2Hz), 2.91 (s, 3H), 2.86 (s, 3H), 2.40 (ddd, 1H, J=13.6,7.8,1.7Hz), 2.32 (ddd, 1H, J=13.7,9.7,9.7Hz) ppm；HRMS(ES⁺)：C₁₂H₁₉N₇O₃Calculated value is 309.1549, and actual value is 310.1633(MH⁺)。

BMA.6：By trichloroethyl chloroformate (49mL, 0.35mmol) be added drop-wise to ice-cold BMA.1 (200mg, In 3.5mL pyridine solutions 0.35mmol).The solid of adhesivity, its slow mechanism dissolved are formed immediately.Mixture 10min is stirred, Add second part of trichloroethyl chloroformate (49mL, 0.35mmol, 1.0 equivalent).Mixture 20min is further stirred at 0 DEG C. Add 10mL NaHCO₃The reaction is quenched in saturated aqueous solution.Mixture is transferred to the CH equipped with 10mL₂Cl₂The leakage of point liquid In bucket.Organic phase is collected, and with 3x 10mL CH₂Cl₂Aqueous layer extracted.The organic fraction of merging is dried with magnesium sulfate, and filtering is simultaneously It is concentrated under reduced pressure as white solid.With silica gel chromatography (94: 6 CH₂Cl₂/ MeOH) material is purified, BMA.6 is obtained, is that white is solid Body (244mg, 93%).TLC R_f=0.34 (9: 1 CH₂Cl₂/MeOH)；¹H NMR(CD₃CN, 500MHz, 70 DEG C) δ 7.76- 7.70 (m, 4H), 7.02-6.94 (m, 4H), 6.78 (br s, 1H), 6.16 (s, 2H), 5.62 (d, 1H, J=7.0Hz), 4.87 (d, 1H, J=12.0Hz), 4.82 (d, 1H, J=12.0Hz), 4.82-4.76 (m, 1H), 4.64 (br s, 1H), 4.61 (dd, 1H, J=7.5,7.5Hz), 4.31 (dd, 1H, J=18.0,11.0Hz), 4.28 (ddd, 1H, J=11.5,11.5,3.5Hz), 4.00-3.92 (m, 1H), 3.85 (s, 3H), 3.84 (s, 3H), 3.48-3.34 (m, 2H), 2.74-2.64 (m, 1H), 2.19- 2.12 (m, 1H) ppm；IR (film) v 3333,1764,1597,1531,1499,1255,1131,1081cm^-1；HRMS(ES⁺)： C₂₆H₃₁N₆O₉S₂Calculated value is 740.0481, and actual value is 763.0546 (MNa⁺)。

BMA.7：By diisopropylethylamine (535mL, 3.1mmol, 10.0 equivalent) be added to BMA.6 (228mg, In 6.0mL MeCN suspension 0.31mmol), and mixture 12h is stirred at 60 DEG C, solid matter dissolves during this period. The reaction is cooled to room temperature, and solution decompression is concentrated, beige solid is produced.In 5mL Et₂Will be not purified in the presence of O Material grind；It is collected by filtration the beige solid, and with Et ice-cold 5mL₂O is washed, and obtains 2.54 (156mg, 86%).TLC R_f=0.34 (9: 1 CH₂Cl₂/MeOH)；¹H NMR(CD₃CN, 500MHz) δ 7.80 (dd, 2H, J=7.0,2.0Hz), 7.70 (dd, 2H, J=7.0,2.0Hz), 7.04 (dd, 2H, J=7.0,2.0Hz), 6.95 (dd, 2H, J=7.0,2.0Hz), 6.25 (br s, 2H), 5.66 (br s, 1H), 4.73 (ddd, 1H, J=14.5,11.0,5.0Hz), 4.66 (br s, 1H), 4.61 (dd, 1H, J=11.0,7.5Hz), 4.38 (dd, 1H, J=8.0,8.0Hz), 4.22 (ddd, 1H, J=10.0,10.0, 1.5Hz), 4.14 (dd, 1H, J=9.0,2.5Hz), 3.85 (s, 3H), 3.82 (s, 3H), 3.61 (ddd, 1H, J=14.5, 7.5,3.0Hz), 3.40 (m, 1H), 2.55-2.46 (m, 1H), 2.23-2.17 (m, 1H) ppm；IR (film) v 3326,3307, 1776,1596,1533,1499,1398,1259,1138,1084cm^-1；HRMS(ES⁺)：C₂₄H₂₈N₆O₈S₂Calculated value is 592.1410, actual value is 615.1308 (MNa⁺)。

BMA.8：Solid KMnO is added into ice-cold BMA.7 (20mg, 0.034mmol) 1mL solution of acetic anhydride₄ (11mg, 0.067mmol, 2.0 equivalent).The bright purple solution 4h is stirred at 0 DEG C.Then 2mL Na is added₂S₂O₃Saturation water The reaction is quenched in solution, and the H of the EtOAc and 5mL with 5mL₂O dilutes.Mixture is transferred in separatory funnel and collected Organic layer.With 3x 5mL EtOAc aqueous layer extracteds.The organic layer of merging is washed with 10mL NaCl saturated aqueous solutions, sulfuric acid is used Magnesium is dried, and is filtered and is concentrated under reduced pressure, obtains oily residue.With silica gel chromatography (95: 5 CH₂Cl₂/ MeOH) material is purified, Acetate BMA.8 is obtained, is white solid (12mg, 53%).TLC R_f=0.40 (9: 1 CH₂Cl₂/MeOH)；¹H NMR (CD₃CN, 400MHz) δ 8.23 (br s, 1H), 7.84-7.80 (m, 2H), 7.75-7.71 (m, 2H), 7.06-7.03 (m, 2H), 6.98-6.94 (m, 2H), 6.48 (br s, 2H), 6.04 (d, 1H, J=6.0Hz), 5.18 (d, 1H, J=4.0Hz), 4.65 (ddd, 1H, J=8.8,7.2,2.4Hz), 4.48 (s, 1H), 4.46 (d, 1H, J=1.2Hz), 4.35 (ddd, 1H, J=6.4, 4.0,2.8Hz), 3.85 (s, 3H), 3.83-3.78 (m, 1H), 3.82 (s, 3H), 3.56-3.50 (m, 1H), 2.66 (dd, 2H, J =6.8,5.2Hz), 2.07 (s, 3H) ppm；IR (film) v 3447,3356,1783,1720,1621,1597,1525,1500, 1399,1260,1138,1082,836cm^-1；HRMS(ES⁺)：C₂₅H₂₈N₆O₁₂S₂Calculated value is 668.1207, and actual value is 689.1312(MNa⁺)。

BMA.9：To ice-cold BMA.7 (20mg, 34.0 μm of ol) 976 50: 10: 1 acetone of μ L/H₂In O/AcOH solution Add KMnO₄(11mg, 67.0 μm of ol, 2.0 equivalents).Stirred at 0 DEG C after 2.5h, add 2mL Na₂S₂O₃Saturated aqueous solution The reaction is quenched.With 5mL EtOAc and 5mL H₂O dilutes the mixture, and is transferred in separatory funnel.Collected organic layer And with 3x 5mL EtOAc aqueous layer extracteds.By NaHCO of the organic layer of merging with 3x 10mL₃Saturated aqueous solution and 1x 10mL NaCl saturated aqueous solutions washing, dried and be concentrated under reduced pressure with magnesium sulfate.The material isolated with Silica gel chromatography, is obtained Keto-alcohol BMA.9, is white solid (11mg, 54%).TLC R_f=0.36 (9: 1CH₂Cl₂/M_eOH)；¹H NMR(CD₃CN, 500MHz) δ 8.18 (br s, 1H), 7.81-7.78 (m, 2H), 7.76-7.73 (m, 2H), 6.43 (br s, 2H), 6.03 (d, 1H, J=6.0Hz), 4.57 (dd, 1H, J=6.0,6.0Hz), 4.50 (ddd, 1H, J=6.0,6.0,2.0Hz), 4.47-4.42 (m, 1H), 4.32-4.29 (m, 1H), 4.24 (ddd, 1H, J=6.5,5.0,1.5Hz), 3.90 (ddd, 1H, J=14.5,9.0, 3.0Hz), 3.85 (s, 3H), 3.82 (s, 3H), 3.57 (ddd, 1H, J=15.0,6.5,4.5Hz), 2.89 (ddd, 1H, J= 14.0,6.0,3.0Hz), 2.59 (ddd, 1H, J=14.0,9.5,4.0Hz) ppm；IR (film) v 3363,1784,1691, 1614,1539,1403,1262,1135,1075,833,763cm^-1。

BMA.10：A oxone (436mg, 0.71mmol, 7.0 equivalent) is added to OsCl₃(36mM's H₂O solution, 282mL, 0.010mmol, 0.10 equivalent) and Na₂CO₃The 3: 3 of the 4.3mL of (107mg, 1.0mmol, 10 equivalent): 1EtOAc/MeCN/H₂In O mixtures.It was observed that there is gas leniently to discharge, the beige coloured suspension mixture of gained is stirred 2min is mixed, then Jia Ru oxazolidones BMA.9 (60mg, 0.10mmol).It is stirred vigorously content 48h.Add 5mL saturation Na₂S₂O₃The reaction is quenched, mixture is then transferred to the H containing 10mL₂In O and 20mL EtOAc separatory funnel.Receive Collect organic layer, and with 3x 15mL EtOAc aqueous layer extracteds.By the organic extract of merging 10mL NaCl saturated aqueous solutions Washing, is dried and is concentrated under reduced pressure with magnesium sulfate.With silica gel chromatography (92: 8 CH₂Cl₂/ MeOH) pure solid residue, obtain BMA.10, is white solid (28mg, 44%).TLC R_f=0.21 (9: 1CH₂Cl₂/MeOH)；¹H NMR(CD₃CN, 500MHz) δ 7.80-7.76 (m, 4H), 7.01-6.95 (m, 4H), 6.51 (br s, 2H), 6.07 (br s, 1H), 4.83 (br s, 1H), 4.46-4.40 (m, 2H), 4.32 (br s, 1H), 4.12 (d, 1H, J=6.0Hz), 3.92 (d, 1H, J=3.5Hz), 3.83 (s, 3H), 3.81 (s, 3H), 3.81-3.75 (m, 1H), 3.44 (ddd, 1H, J=12.0,12.0,2.0Hz), 2.25 (¹H, signal quilt DHO is covered), 1.84 (dd, 1H, J=12.5,7.5Hz) ppm；IR (film) v 3326,3307,1776,1596,1533, 1499,1398,1259,1138,1084cm^-1；HRMS(ES⁺)：C₂₄H₂₈N₆O₁₀S₂Calculated value is 624.1308, and actual value is 625.1359(MH⁺)。

BMA.11：To 2: 2: 1EtOAc/MeCN/H of ice-cold BMA.9 (30mg, 51.0 μm of ol) 2.5mL₂In O solution Add RuCl₃(0.53mg, 2.5 μm of ol, O.05 equivalent), then add solid NaIO₄(13.1mg, 61.2mmol, 1.2 equivalent). Stirring carries out reaction up to 45min at 0 DEG C, then adds 2mL Na₂S₂O₃The reaction is quenched in saturated aqueous solution.With 5mL's EtOAc and 5mL H₂O dilutes the reaction, and is transferred in separatory funnel.Collected organic layer is simultaneously extracted with 3x 5mL EtOAc Water layer.The organic fraction of merging is washed with 10mL NaCl saturated aqueous solutions, dried with magnesium sulfate, is filtered, and be concentrated under reduced pressure. The material isolated is purified with silica gel chromatography, glycol BMA.11 is obtained, is white solid (23mg, 72%).TLC R_f=0.30 (9: 1CH₂Cl₂/MeOH)；¹H NMR(CD₃CN, 400MHz) δ 7.90 (br s, 1H), 7.82-7.79 (m, 2H), 7.76-7.73 (m, 2H), 7.06-7.02 (m, 2H), 7.00-6.96 (m, 2H), 6.36 (br s, 2H), 5.67 (br d, 1H, J =3.6Hz), 4.57-4.52 (m, 1H), 4.31 (dd, 1H, J=11.2,5.6Hz), 4.30-4.27 (m, 1H), 3.88-3.79 (m, 1H), 3.84 (s, 3H), 3.82 (s, 3H), 3.73-3.64 (m, 2H), 3.45-3.37 (m, 1H), 3.32-3.17 (m, 2H), 1.91-1.81 (m, 1H), 1.70-1.61 (m, 1H) ppm；IR (film) v 3325,1767,1618,1596,1534,1499, 1400,1260,1134,1083cm^-1。

Oxazolidone BMA.10 is converted into by STX derivatives BMA.14-BMA.18 using general three-step approach.By BMA.10 The Experimental detail for being converted into BMA.14 is representative.

BMA.12：The tert-butyl group -6- amino cyclohexyl aminos formic acid esters (28mg, 0.13mmol, 5.0 equivalent) is added into BMA.10 In the 1.3mL THF solutions of (16mg, 26.0 μm of ol).Mixture 4h is stirred, is concentrated under reduced pressure, with silica gel chromatography (94: 6 CH₂Cl₂/ MeOH) material that is isolated and purified with, BMA.12 is obtained, is colourless oil (22mg, 99%).TLC R_f=0.30 (9: 1 CH₂Cl₂/MeOH)；¹H NMR(CD₃CN, 500MHz, 60 DEG C) δ 7.75 (dd, 4H, J=9.0,1.5Hz), 6.98 (dd, 4H, J= 9.0,2.5Hz), 6.37 (br s, 2H), 5.81 (br s, 1H), 5.53 (br s, 1H), 5.11 (br s, 1H), 4.81 (br s, 1H), 4.29 (br d, 1H, J=11.5Hz), 4.13 (s, 1H), 4.03 (br t, 1H, J=8.0Hz), 3.94 (t, 1H, J= 3.5Hz), 3.85 (s, 6H), 3.75 (dd, 1H, J=12.0,4.5Hz), 3.67-3.63 (m, 1H), 3.58-3.50 (m, 2H), 3.07 (br s, 2H), 2.99 (ddd, 2H, J=6.5,6.5,6.5Hz), 2.23-2.16 (m, 1H), 1.85-1.81 (m, 1H), 1.51-1.40 (m, 4H), 1.42 (s, 9H), 1.34-1.25 (m, 4H) ppm；IR (film) v 3330,2932,1701,1578, 1535,1499,1256,1132,1082cm^-1；HRMS(ES⁺)：C₃₅H₅₂N₈O₁₂S₂Calculated value is 840.3146, and actual value is 863.3033(MNa⁺)。

BMA.13：The 10mL round-bottomed flasks that will be equipped with BMA.12 (23mg, 0.027mmol) are put in ice bath, slowly to it Middle addition B (O₂CCF₃)₃(0.5M CF₃CO₂H solution, 1.64mL, 0.82mmol, 30 equivalents).Light brown obtained by stirring is molten Liquid, room temperature is to slowly warm up to through 5h.Continue to heat after 14h at such a temperature, the solution is cooled to 0 DEG C, and be added dropwise 1.0mL's The reaction is quenched in MeOH.Be concentrated under reduced pressure the mixture, obtains oily residue.Not purified product is re-dissolved in~2mL MeOH in, and concentrate the solution.Said process is repeated once.Then the material isolated is dissolved in 1mL H₂O, and make By 2x 10cm Dowex 1x 8-200 (^-OH types) post.The fraction containing product that (＞ 7.5) is determined by pH is collected, and It is acidified with 100mL 1.0M HCl/waters solution.Solutions in Freeze-drying, obtains BMA.13, is white powder (12mg, 92%).¹H NMR (D₂O, 400MHz) δ 4.77 (d, 1H, J=1.2Hz), 4.33 (d, 1H, J=3.6Hz), 4.26 (dd, 1H, J=11.6, 9.2Hz), 4.01 (dd, 1H, J=11.6,5.6Hz), 3.81 (dd, 1H, J=9.2,5.6Hz), 3.77 (ddd, 1H, J= 10.0,10.0,2.0Hz), 3.67 (ddd, 1H, J=18.8,8.8,1.6Hz), 3.15-3.05 (m, 2H), 2.97 (dd, 2H, J =7.6,7.6Hz), 2.46-2.36, (m, 1H), 2.23 (ddd, 1H, J=14.8,8.4,1.6Hz), 1.68-1.61 (m, 2H), 1.52-1.45 (m, 2H), 1.41-1.31 (m, 4H) ppm；HRMS(ES⁺)：C₁₆H₃₀N₈O₃Calculated value is 382.2441, actual value For 383.2514 (MH⁺)。

BMA.14：Powdery is added into BMA.13 (9mg, 0.018mmol) 1.4mL DMSO solutionsMolecular sieve.Stir Suspension 20min is mixed, dicyclohexylcarbodiimide (45mg, 0.22mmol, 12 equivalent) and pyridinium trifluoroacetate is then added Salt (27mg, 0.14mmol, 7.5 equivalent).It is immediately generated white precipitate；The slurry is stirred vigorously up to 17h.Lyophilized reaction mixing Thing, obtains solid product, is resuspended in 1mL H₂In O, and filtered with the thin pad of diatomite.In addition with 2x 1mL H₂O is to protect Demonstrate,prove the quantitative transfer of the material.The filtrate of merging is freezed, and with the reverse HPLC-purified solid product (Altima isolated C18,10 μm, 10x 250mm posts, 20: 80 MeCN/10mM C₃F₇CO₂The H aqueous solution → 27: 73 MeCN/10mM C₃F₇CO₂H water Solution gradient elutes 14min, 214nm UV detections).Flow velocity is 6mL/min, and BMA.14 retention time is 7.1min, freezes and divides From product is white powder (12mg, 63%).¹H NMR(D₂O, 500MHz) δ 4.68 (s, 1H), 4.23 (dd, 1H, J=11.5, 9.5Hz), 3.97 (dd, 1H J=11.5,5.5Hz), 3.78-3.74 (m, 2H), 3.52 (ddd, 1H, J=18.5,8.5, 1.5Hz), 3.09-3.01 (m, 2H), 2.93 (dd, 2H, J=6.4,6.4Hz), 2.38 (ddd, 1H, J=14.0,8.0, 2.0Hz), 2.33-2.26 (m, 1H), 1.63-1.57 (m, 2H), 1.48-1.42 (m, 2H), 1.35-1.28 (m, 4H) ppm； HRMS(ES⁺)：C₁₆H₃₀N₈O₄Calculated value is 398.2390, and actual value is 399.2472 (MH⁺)。

BMA.15：¹H NMR(D₂O, 400MHz) δ 4.70 (s, 1H), 4.27 (dd, 1H, J=11.6,8.8Hz), 3.99 (dd, 1H, J=11.6,5.2Hz), 3.80-3.76 (m, 2H), 3.54 (dd, 1H, J=7.6,7.6Hz), 3.11-3.04 (m, 2H), 2.39-2.30 (m, 2H), 1.45 (dd, 2H, J=6.8,6.8Hz), 1.26-1.21 (m, 8H), 0.83 (t, 3H, J= 6.8Hz)ppm；HRMS(ES⁺)：C₁₇H₃₁N₇O₄Calculated value is 397.2438, and actual value is 398.2505 (MH⁺)。

BMA.16：¹H NMR(D₂O, 500MHz) δ 4.70 (s, 1H), 4.26 (dd, 1H, J=11.5,9.0Hz), 3.99- 3.95 (m, 1H), 3.80-3.75 (m, 2H), 3.66-3.60 (m, 1H), 3.54 (ddd, 1H, J=18.0,10.0,1.5Hz), (d, 6H, the J=7.0Hz) ppm of 2.40 (ddd, 1H, J=14.0,8.0,1.5Hz), 2.34-2.28 (m, 1H), 1.09；HRMS(ES⁺)：C₁₃H₂₃N₇O₄Calculated value is 341.1812, and actual value is 342.1890 (MH⁺)。

BMA.17：¹H NMR(D₂O, 500MHz) δ 4.69 (d, 1H, J=1.5Hz), 4.27 (dd, 1H, J=12.0, 9.5Hz), 3.97 (dd, 1H, J=11.5,5.0Hz), 3.78-3.71 (m, 2H), 3.52 (dd, 1H, J=9.0,9.0Hz), 3.08 (dd, 2H, J=6.5,6.5Hz), 2.15 (t, 2H, J=7.0Hz), 1.55-1.49 (m, 2H), 1.49-1.43 (m, 2H), 1.31-1.23 (m, 2H) ppm (notes：Due to D₂O is exchanged, so without the methenes of C 11¹H signal)；HRMS(ES⁺)： C₁₆H₂₇N₇O₆Calculated value is 413.2023, and actual value is 414.2122 (MH⁺)。

BMA.18：¹H NMR(D₂O, 400MHz) δ 7.87 (d, 2H, J=9.2Hz), 7.85 (d, 2H, J=9.2Hz), 7.81 (dd, 2H, J=8.0,1.2Hz), 7.73 (tt, 1H, J=7.6,1.6Hz), 7.57 (t, 2H, J=8.0Hz), 4.64 (d, 1H, J =1.2Hz), 4.17 (dd, 1H, J=11.6,9.6Hz), 3.96 (dd, 1H, J=11.6,4.8Hz), 3.74-3.69 (m, 2H), 3.53 (dd, 2H, J=5.2,5.2Hz), 3.41-3.34 (m, 3H), 2.34-2.26 (m, 2H) ppm；HRMS(ES⁺)： C₂₆H₃₀N₈O₆Calculated value is 550.2288, and actual value is 551.2388 (MH⁺)。

BMA.20：To BMA.13 (2.5mg, 2.4mmol) 240 μ L the buffer solutions of pH 9.5 (0.1M NaHCO₃/ Na₂CO₃) and DMF for added in the solution in 1: 3 mixture 4- fluobenzoic acids N-hydroxy-succinamide esters (1.6mg, 7.2mmol, 3.0 equivalents).Solution 5h is stirred at room temperature, is then acidified with 30mL 1.0M HCl/waters solution.This is freezed to mix Compound, obtains solid matter, with reverse HPLC-purified (Altima C18,10 μm, 10x 250mm posts, 10: 90 MeCN/10mM C₃F₇CO₂The H aqueous solution → 40: 60MeCN/10mM C₃F₇CO₂The H aqueous solution gradient elution 30min, 254nm UV are detected).Flow velocity is 6mL/min, BMA.20 retention time are 16.2min, freeze separation, and product is white powder (2.2mg, 96%).¹H NMR (D₂O, 500MHz) δ 7.75-7.72 (m, 2H), 7.21-7.18 (m, 2H), 4.68 (s, 1H), 4.21 (dd, 1H, J=12.0, 9.5Hz), 3.84 (dd, 1H, J=12.0,5.5Hz), 3.78-3.74 (m, 2H), 3.54-3.49 (m, 1H), 3.34 (dd, 2H, J =7.0,7.0Hz), 3.10-3.03 (m, 2H), 2.40-2.26 (m, 2H), 1.61-1.55 (m, 2H), 1.48-1.43 (m, 2H), (1.38-1.28 m, 4H) ppm；HRMS(ES⁺) calculated value be C₂₃H₃₃FN₈O₅520.2558, actual value is 521.2639 (MH⁺)。

TLC R_f=0.08 (9: 1 CH₂Cl₂/MeOH)；¹H NMR(CDCl₃, 400MHz) and δ 7.91 (d, 2H, J=8.0Hz), 7.81-7.75 (m, 4H), 7.43 (d, 2H, J=8.4Hz), 6.78 (br s, 1H), 4.63 (br s, 1H), 3.96 (s, 2H), 3.45 (q, 2H, J=6.8Hz), 3.11 (q, 2H, J=6.4Hz), 1.66-1.57 (m, 4H), 1.51-1.32 (m, 4H) ppm；IR (film) v 3361,3305,2931,2859,1686,1648,1523,1280,1174,932cm^-1。

BMA.23：To the BMA.22 (4.0mg, 4.1 μm of ol) of stirring in the 200 μ L buffer solution of MeCN and pH 9.5 (NaHCO₃/Na₂CO₃) for 1: 1 mixture in solution in add biotin NHS esters (2.1mg, 6.2 μm of ol, 1.5 equivalents). Mixture 6h is stirred, then adding 21 μ L 1.0M HCl/water solution is quenched the reaction, decompression removes solvent.With anti-phase Material (18,10 μm of Altima C, 10x 250mm posts, 20: 80 MeCN/H that HPLC is isolated and purified with₂O (contains 0.1% CF₃CO₂The H aqueous solution) → 80: 20MeCN/H₂O (contains 0.1%CF₃CO₂The H aqueous solution) gradient elution 30min, 254nm UV detections). Flow velocity is 6mL/min, and BMA.23 retention time is 8.9min, freezes separation, and product is white powder (0.71mg, 20%).¹H NMR(D₂O, 400MHz) δ 7.84 (s, 4H), 7.79 (d, 2H, J=8.0Hz), 7.46 (d, 2H, J=7.6Hz), 4.72 (s, 1H), 4.47 (dd, 1H, J=7.6,5.2Hz), 4.40 (d, 2H, J=6.0Hz), 4.31-4.27 (m, 2H), 4.05 (dd, 1H, J =12.0,5.2Hz), 3.83-3.74 (m, 2H), 3.56-3.49 (m, 1H), 3.39 (t, 2H, J=6.8Hz), 3.21-3.13 (m, 3H), 2.85 (dd, 1H, J=12.8,4.8Hz), 2.63 (d, 1H, J=13.2Hz), 2.42-2.26 (m, 2H), 2.14 (t, 2H, J=7.2Hz), 1.65-1.57 (m, 4H), 1.55-1.44 (m, 4H), 1.39-1.26 (m, 6H) ppm.

¹H NMR(D₂O, 400MHz) δ 4.68 (s, 1H), 4.28 (dd, 1H, J=11.6,9.6Hz), 3.96 (dd, 1H, J= 12.0,5.2Hz), 3.78 (ddd, 1H, J=10.0,10.0,2.0Hz), 3.77-3.74 (m, 1H), 3.54 (ddd, 1H, J= 18.4,10.4,2.8Hz), 3.15 (s, 2H), 2.39ddd, 1H, J=14.4,8.4,2.0Hz), 2.30 (ddd, 1H, J= 14.4,10.0,10.0Hz), 1.00 (s, 3H) ppm；HRMS(ES⁺)：C₁₃H₂₁N₉O₄Calculated value is 367.1717, and actual value is 368.1789(MH⁺)。

BMA.25：To the BMA.13 (2.6mg, 2.5 μm of ol) of stirring in buffer solution (the 0.1M NaHCO of 125 μ L pH 9.5₃/ Na₂CO₃The aqueous solution) in solution in add Oregon Green NHS esters (4.0mg, 7.8 μm of ol, 3 equivalents) 125 μ L MeCN Solution.Mixture 4h is stirred at room temperature, is then acidified with 37.5 μ L 1.0M HCl/waters solution.Be concentrated under reduced pressure the solution, is used in combination Reverse HPLC-purified solid matter (Altima C18,10 μm, 10x 250mm posts, 10: 90 MeCN/10mM C₃F₇CO₂H is water-soluble Liquid → 40: 60 MeCN/10mM C₃F₇CO₂The H aqueous solution gradient elution 30min, 254nih UV are detected).Flow velocity is 6mL/min. BMA.25 retention time is 28.5min, freezes separation, and product is orange solids (0.37mg, 19%)：¹H NMR(D₂O, 400MHz) δ 8.47 (d, 1H, J=1.6Hz), 8.10 (dd, 1H, J=8.0,1.6Hz), 7.44 (d, 1H, J=8.0Hz), 7.00 (s, 1H), 6.99 (s, 1H), 6.91 (s, 1H), 6.88 (s, 1H), 4.66 (s, 1H), 4.19 (dd, 1H, J=11.6,9.2Hz), 3.95 (dd, 1H, J=11.6,5.2Hz), 3.78-3.71 (m, 2H), 3.50 (dd, 1H, J=18.4,10.0Hz), 3.43 (t, 2H, J=6.8Hz), 3.11-3.03 (m, 2H), 2.36 (ddd, 1H, J=14.0,8.4,1.6Hz), 2.28 (ddd, 1H, J= 14.0,9.6,9.6Hz), 1.68-1.61 (m, 2H), 1.51-1.44 (m, 2H), 1.41-1.34 (m, 4H) ppm；HRMS(ES⁺)： C₃₇H₃₈F₂N₈O₁₀Calculated value is 792.2679, and actual value is 793.2753 (MH⁺)。

BMA.26：To the BMA.13 (7.0mg, 6.7 μm of ol) of stirring in the buffer solution (0.1M of 340 μ L MeCN/pH 9.5 NaHCO₃/Na₂CO₃The aqueous solution) to add Cy5-NHS esters in the solution in 1: 1 mixture, (6.2mg, 10 μm of ol, 1.5 work as Amount).Mixture 4h is stirred at room temperature, is then acidified with 68 μ L 1.0M HCl/waters solution.Be concentrated under reduced pressure the solution, and with anti-phase (Altima C18,10 μm, 10x 250mm posts, 20: 80 MeCN/ contain 0.1%CF to HPLC pure solids material₃CO₂H H₂O→ 50: 50 MeCN/ contain 0.1%CF₃CO₂H H₂O gradient elutions 30min, 254nm UV are detected).Flow velocity is 6mL/min.BMA.26 Retention time be 26.0min, freeze separation, product be blue solid (1.03mg, 18%)：¹H NMR(D₂O, 400MHz) δ 8.01-7.94 (m, 2H), 7.50-7.47 (m, 2H), 7.41-7.37 (m, 2H), 7.24 (t, 4H, J=7.6Hz), 6.51 (t, 1H, J=12.8Hz), 6.23 (d, 1H, J=2.4.hz), 6.20 (d, 1H, J=2.4Hz), 4.64 (d, 1H, J=1.2Hz), 4.10 (dd, 1H, J=11.6,9.2Hz), 4.97 (t, 2H, J=6.4Hz), 3.81 (dd, 1H, J=11.6,5.6Hz), 3.75 (ddd, 1H, J=10.0,10.0,2.4Hz), 3.69 (dd, 1H, J=9.2,6.0Hz), 3.54 (s, 3H), 3.48 (dd, 1H, J =18.0,10.0Hz), 3.00-2.93 (m, 4H), 2.37 (ddd, 1H, J=14.0,8.4,2.4Hz), 2.29 (ddd, 1H, J= 13.6,10.4,10.4Hz), 2.15 (t, 2H, J=6.4Hz), 1.85-1.78 (m, 2H), 1.62 (s, 6H), 1.62 (s, 6H), 1.61-1.54 (m, 2H), 1.37-1.24 (m, 6H), 1.20-1.16 (m, 2H) ppm；HRMS(ES⁺)：C₄₈H₆₇N₁₀O₅ ⁺Calculated value For 863.5290, actual value is 430.7580 (MH⁺/2)。

BMA.27：To the BMA.13 of stirring in the buffer solutions of MeCN/pH 9.5 (0.1M NaHCO₃/Na₂CO₃The aqueous solution) be 1: DCDHF-NHS esters are added in solution in 1 mixture.Lower progress reaction is stirred at room temperature and reaches 3h, then with 40 μ L 1.0M HCl/water solution is acidified.It is concentrated under reduced pressure after the solution, with reverse HPLC-purified solid matter (Altima C18,10 μm, 10x 250mm posts, 20: 80 MeCN/ contain 0.1%CF₃CO₂H H₂O → 80: 20 MeCN/ contain 0.1%CF₃CO₂H H₂O gradient elutions 30min, 254nm UV are detected).Flow velocity is 6mL/min.BMA.27 retention time is 14.6min, freezes separation, and product is deep Violet solid (1.3mg, 44%)：¹H NMR(D₂O, 500MHz) δ 7.73 (d, 1H, J=16.5Hz), 7.55 (d, 2H, J= 8.0Hz), 6.82 (d, 2H, J=8.5Hz), 6.57 (d, 1H, J=15.5Hz), 4.68 (s, 1H), 4.20 (dd, 1H, J= 11.0,11.0Hz), 3.96 (dd, 1H, J=10.5,4.5Hz), 3.78-3.73 (m, 2H), 3.53-3.48 (m, 3H), 3.08 (s, 3H), 3.01-2.99 (m, 4H), 2.38 (dd, 1H, J=14.0,5.6Hz), 2.33-2.25 (m, 3H), 1.97-1.92 (m, 2H), 1.41-1.36 (m, 2H), 1.33-1.28 (m, 2H), 1.22-1.17 (m, 4H) ppm.

WHP.3：To WHP.1 (1.5mg, 0.0015mmol) in 700 8.5 buffer solution (0.1M of μ L MeCN and pH NaH₂PO₄/Na₂HPO₄) for 2: 1 mixture in solution in add 3- maleimidopropionic acid N-hydroxy-succinamide esters (2.1mg, 0.0077mmol, 5.0 equivalent).Mixture 4h is stirred at room temperature, is then acidified with 50 μ L 1.0M HCl/waters solution. Freeze the mixture, obtain solid matter, then with it is reverse HPLC-purified (Altima C18,10 μm, 10x 250mm posts, 20: 80 MeCN/10mM C₃F₇CO₂The H aqueous solution → 27: 73 MeCN/10mM C₃F₇CO₂The H aqueous solution gradient elutions 14min, 214nm UV is detected).Flow velocity is 6mL/min, and WHP.3 retention time is 7.1min, freezes separation, product be white powder (0.38mg, 28%).¹H NMR(D₂O, 500MHz) δ 6.81 (s, 2H), 4.60 (s, 1H), 4.31 (dd, 1H, J=11.3,9.5Hz), 3.96 (dd, 1H, J=12.0,5.0Hz), 3.81-3.76 (m, 2H), 3.75 (t, 2H, J=6.0Hz), 3.58-3.54 (m, 1H), 3.31-3.20 (m, 2H), 3.14 (t, 2H, J=10.5Hz), 2.47 (t, 2H, J=6.5Hz), 2.41-2.29 (m, 2H) ppm； LRMS(ES⁺)C₁₉H₂₇N₉O₇Calculated value is 493.47, and actual value is 494.59 (MH⁺)。

Embodiment 2：

The synthesis path of saxitoxin analog and correlation molecule is present embodiments provided, with GTX-3 (GTX3) exemplified by synthesis, it is with similar to other saxitoxin analog (such as saxitoxins, N-STX With other gonyatoxins) paralytic shellfish poisoning (PSP)s of the small molecule biguanides based structures of structure.Known sulfuric acid more than 20 kinds Detoxification element --- the first synthesis path of any one in GTX-3 (GTX3) is described (figure in the following literature 9).(referring to document：(a) Shimizu, Y.；Buckley, L.J.；Alam, M.；Oshima, Y.；Fallon, W.E.；Kasai, H.；Miura, I.；Gullo, V.P.；Nakanishi, K.J.Am.Chem.Soc.1976,98,5414-5416.(b) Boyer, G.L.；Schantz, E.J.；Schnoes, H.K.J.Chem.Soc., Chem.Comm.1978,889-890..(c) Onodera, H.；Satake, M.；Oshima, Y.；Yasumoto, T.；Carmichael, W.W.Natural Toxins 1997,5,146- 151.Documents below describes GTX 2 and 3 endless total synthesis method：Hannick, S.M.；Kishi, Y.J.Org.Chem.1983,48,3833-3835.)

We have recently disclosed the method for oxidation for forming 2- aminooimidazoles, the five-membered ring guanidine in GTX 3 turns into afterwards The focus of our synthesis analysis.(referring to document：Kim, M.；Mulcahy, J.V.；Espino, C.G., Du Bois, J.Org.Lett.2006,8,1073-1076.) this conversion is considered as that the guanidine nitrene that is combined by Rh (can modify c h bond With the active material of both pi bonds) mediation., will by guanidine nitrence (guanidine nitrenoid) in order to prepare GTX 3 Pyrrole nucleus ammonification is the new opplication (Fig. 9) of the technology.Fig. 9 shows oxidizable pyrrole, and highlights GTX3 synthetic method.This is anti- Be able to should occur by tension force ethylene imine 3 or dipole thing 4, wherein C10 or C12 can be produced required three rings mother by nucleophillic attack Core.(the similar oxidation reaction with indole derivatives gives the evidence of zwitterionic intermediate, referring to document：Padwa, A.； Flick, A.C.；Leverett, C.A.；Stengel, T.J.Org.Chem.2004,69,6377-6386.) despite the presence of the area Domain chemistry (regiochemical) problem, but GTX problem reductions are quite unremarkable bicyclic intermediate by such strategy 1.According to the method, the approach of such synthesis biguanides 1 is formulated, it uses pyrroles and the intramolecular addition of active imines.Though Right precedent is limited, but is due to that necessary precursor 2 is easy to obtain from serine, it is possible to which Fast Evaluation is such Pictet-Spengler reacts.

GTX 3 synthesis is since the three step transforming sequences that 1- serine methylesters are converted into aldehyde 5 (Figure 10).(referring to text Offer：Boger, D.L.；Patel, M.J.Org.Chem.1987,52,2319-2323.) be condensed the aldehyde and allylamine, Ran Houyong BF₃·OEt₂Processing, it can realize the closure of required ring, so that trans substituted with ＞ 20: 1 cis-selectivity generation Urea 6.(x-ray crystal structure of the modified forms of the intermediate confirms that the trans geometric is arranged.) assume the C5/C6 of the product Stereochemical structure (GTX numberings) is set up under kinetic control, then reduces the pi-allyl between the substituent on C6 and N7 The conformation of power is probably the reason for causing observed diastereoselectivity.By four step transforming sequences effectively by 6 It is converted into necessary ammonification precursor 7；It is noted that a one-step process is developed, for sequentially carrying out allyl deprotection protection and different sulphur The formation of urea is (referring to step e, 6 → 7, Tces=SO₃CH₂CCl₃)。

The Rh Study on Catalytic Amination of Alcohols that guanidine 7 is applied successfully is reacted with unique definition event (singular, defining Event mode) has been assembled into GTX 3 three ring frameworks.The c h bond at the C6 center neighbouring due to being inserted into seem not with pyrroles Modification is competed, therefore the reaction is chemo-selective.The accessory substance acetic acid produced in this transformation is a word used for translation of presumption Third pyridine adds regioselectivity and stereoselectivity, so as to produce unique product N, O- acetals 8 (are based on to reactant mixture 's¹H NMR are analyzed).(this material is in SiO₂On unstability be likely to result in separation yield reduction.The selection of solvent is also right The step is presented with suitable significant impact, CH₂Cl₂Uniquely wherein to observe the medium that starting guanidine 7 is totally consumed.) Although acetate only attacks C10 rather than C12, suitably handle three rings 8 isolated to complete GTX synthesis.

8 are proved to be that some are changeable for the stability of processing and purification process, will so as to promote to make decision Reduce the N of potentially unstable, O- acetal units.In Et₃SiH and BF₃·OEt₂In the presence of the conversion be smoothed out, produce yield For 81% C11-C12 alkene.Change alkene (transposed olefin) product is not detected by under these conditions.Using Cl₃CC (O) NCO makes it possible to introduce primary carbamate.(referring to document：Kocovsky, P.Tetrahedron Lett.1986,27,5521-5524.) intermediate 9 is included in all necessary carbon center for finding in natural products.

Have studied the alternative that alkene 9 is converted into corresponding α -one alcohol.The ketone hydroxylation of regioselectivity Target provides most easily approach for needed for；But not yet determine such condition.(precedent of ketone hydroxylation is referring to text Offer：Fleming, J.J.；McReynolds, M.D.；Du Bois, J.J.Am.Chem.Soc.2007,129,9964-9975.) By contrast, using 2mol% OsO₄With N-methylmorpholine-N- oxides carry out olefin-dihydroxy be it is fairly effective, And glycol 10 is produced, it is single stereoisomer.Molecular modeling assay shows that β-face of the alkene in 9 is more exposed , this is consistent with the selectivity observed.Under conditions of the height optimization using benzoyl cyanide and DMAP, C11- is realized OH protection.In addition, more standard acylation agent (such as PhC (O) C1) and 3.Amine or the combination of pyridine base, are produced inseparable The mixture of isomeric benzoylate.Although other blocking groups can be installed on C11 such as^tBuMe₂Si-, but they The larger steric bulk being had inhibit C12 alcohol with rear oxidation.Using Dai Si-Martin's oxidant (Dess-Martin Periodinane) can make 11 on C12 positions formed ketone (oxidation protocol based on Cr VI, TEMPO and DMSO can generally disappear Starting material is consumed without generating ketone 12.)

By single stepping remove 12 in all 3 blocking groups, obtained 11 beta-hydroxy saxitoxins, its with Double-C₃F₇CO₂ ^-The form of salt is separated.The material analyze data (¹H NMR, HRMS) it is consistent with described in document.(ginseng See document：(a) Wichmann, C.F.；Boyer, G.L.；Divan, C.L.；Schantz, E.J.；Schnoes, H.K.Tetrahedron Lett.1981,22,1941-1944.(b) Shimizu, Y.；Kobayashi, M.；Genenah, A.； Oshima, Y.Tetrahedron 1984,40,539-544.)

In order to complete GTX 3 synthesis, using DMFSO₃(acid removing is used as with 2,6- di-t-butyl -4- picolines Agent) realize the selectively sulfated of C11 alcohol.Pure toxin is obtained after reversed-phase HPLC, it is C₃F₇CO₂ ^-Addition product.This material It is consistent with the natural GTX 3 of report physical property in every respect, and the Na of heterogenous expression can be blocked_VIn 1.4 ion channels Electrical conduction, its IC₅₀It is worth for~20nM (the IC of report₅₀=13.2-33.5nM).(referring to document：(a) Choudhary, G.； Shang, L.；Li, X.；Dudley, Jr., S.C.Biophys.J.2002,83,912-919.(b) Kao, C.Y.；Kao, P.N.； James-Kracke, M.R.；Koehn, F.E.；Wichmann, C.F.；Schnoes, H.K.Toxicon 1985,23,647- 655.) the GTX 3 pH=8 aqueous solution is stood after, produce GTX 2 in C11 generation epimerisms, this also with it is disclosed Visual report is consistent (Figure 11).(referring to document：For example, (a) Shimizu, Y.；Buckley, L.J.；Alam, M.；Oshima, Y.；Fallon, W.E.；Kasai, H.；Miura, I.；Gullo, V.P.；Nakanishi, K.J.Am.Chem.Soc.1976, 98,5414-5416.(b) Wichmann, C.F.；Boyer, G.L.；Divan, C.L.；Schantz, E.J.；Schnoes, H.K.Tetrahedron Lett.1981,22,1941-1944.) Figure 11 show stand after C11 occur epimerism.

GTX 3 fully synthetic saxitoxin analog and its dependency structure to prepare this family provides the plan of uniqueness The scheme of omiting, while it is however emphasized that effect that Rh Study on Catalytic Amination of Alcohols is assembled for heterocycle.(documents below, which has been described, attempts to guanidine C- H keys insert to assemble the bromine azole derivatives of natural products：Wang, S.；Romo, D.Angew.Chem.Int.Ed.2008,47, 1284-1286.The recent review of this family's molecule can be found in document：M.；Grube, A.；Seiple, I.B.；Baran, P.S.Angew.Chem.Iht.Ed.2007,46,6586-6594.)

Material and method are unless otherwise stated, all reagents are commercially available.Reaction is in drying nitrogen environment Under in the glassware of drying carry out.The liquid of air-sensitive and moisture-sensitive is shifted by syringe or stainless steel pipe And solution.(about 15 support) rotary evaporation concentrates organic solution under reduced pressure.Before it will use, make dichloromethane (CH₂Cl₂), tetrahydrofuran (THF), acetonitrile (MeCN), DMF, acetone (PhMe) pass through activated alumina column Son.Chromatography purifying is carried out to product using forced flow chromatography on Silicycle silica gel 60 (40-63 μm).In EM Science silica gel 60F₂₅₄Thin-layered chromatography is carried out on plate (250 μm).By fluorescent quenching and by using anisaldehyde ethanol (ethanolic anisaldehyde), potassium permanganate solution or ammonium molybdate cerium (CAM) aqueous solution dye to realize expansion The visualization of chromatogram.Using Varian equipment and Alltima C₁₈, 10x 250mm, 10 μm of posts, using containing 0.1% CF₃CO₂H buffer solutions or 10mM C₃F₇COOH MeCN/H₂It is (as follows that O carries out high performance liquid chromatography (HPLC) purifying as eluent Text).

Detected respectively under 400,500 or 600 and 100,125 or 150MHz on Varian Inova spectrometers¹H With¹³C, and it regard residual solvent signal as internal reference.¹H NMR datas record is as follows：Chemical shift (δ, ppm), multiplicity (s, list Peak；D, it is bimodal；T, triplet；Q, quartet；Quint, quintet；M, multiplet；Br, broad peak), integration, coupling constant (Hz) 。¹³C data are with chemical shift (δ, ppm) report.Record uses NaCl salt on Thermo-Nicolet300FT-IR spectrometers The infrared spectrum (IR) of the film of piece, and reported with absorption frequency.It is located at what is worked under Na D lines by the way that sample is loaded In the pond of 50mm on Jasco DIP-1000 digital polarimeters, optically-active data are obtained.High resolution mass spec is big from Stamford Vincent Coates foundation Mass Spectrum Laboratories.

Experimental program and characterize data：

To ice-cold KO^tBu (46.0g, 409.9mmol, 1.1 equivalent) 1.2L 1: 1Et₂It is slowly added in O/THF solution Pyrroles (26.0mL, 372.0mmol).Reactant mixture is heated to room temperature and 30min is stirred.Afterwards, it is slow from flask top Add excessive solid CO₂(~200g), causes violent bubbling, and reaction temperature is reduced.The reaction vessel is put in room temperature In water-bath, and there is no solid CO to drag for standing₂.Add 300mL H₂O, and content is transferred to separatory funnel In.Water layer is collected, and by organic phase 300mL H₂O is washed.The water-phase extract of merging is acidified with 1.0M HCl/waters solution To pH ＜ 1.Then 400mL Et is added into the aqueous solution₂O, and content is transferred again into separatory funnel.Collection has Machine phase, and with 2x 400mL Et₂O aqueous phase extracteds.By the organic extract Na of merging₂SO₄Dry, filtering, and depressurize dense Contracting, obtains pyrroles's -1- carboxylic acids, is white solid (35.6g, 86%).Mp is 114116 DEG C；¹H NMR(CDCl₃, 400MHz) 8.54 (br s, 1H), 7.31 (t, 2H, J=2.4Hz), 6.31 (t, 2H, J=2.4Hz) ppm., itself and number reported in the literature According to consistent.(referring to document：Boger, D.L.；Patel, M. " Activation and coupling of pyrrole-1- carboxylic acid in the formation of pyrrole N-carbonyl compounds-pyrrole-1- Carboxylic acid anhydride. " Journal of Organic Chemistry1987,52,2319-2323.)

To the 1.25L CH of pyrrol-carboxylic acid's (35.6g, 323.3mmol, 2.0 equivalent)₂Cl₂A solid two is added in solution Carbodicyclo hexylimide (68.3g, 331.1mmol, 2.05 equivalent).It is stirred vigorously after mixture 20min, adds portion and grind Very thin 1- serine methyl ester hydrochlorides (25.2g, 161.7mmol) and triethylamine (36.7mL, 242.2mmol, 1.5 equivalent) exist 550mL CH₂Cl₂In suspension.Mixture 22h is stirred for, is then filtered by Celite pad.With cold CH₂Cl₂Washing Flask and filter cake, the filtered fluid of merging is concentrated under reduced pressure.Material (the gradient elution isolated with Silica gel chromatography：7∶1→ 1: 1 hexane/EtOAc), urea JVM.1 is obtained, is lurid oily (22.9g, 67%).TLC R_f=0.24 (1: 1 hexane/ EtOAc)；¹H NMR(CDCl₃, 400MHz) and δ 7.30-7.26 (m, 2H), 7.11-7.05 (m, 1H), 7.24-7.21 (m, 2H), 4.67-4.64 (m, 1H), 4.03 (dd, 1H, J=11.2,4.0Hz), 3.92 (dd, 1H, J=11.6,3.6Hz), 3.73 (s, 3H)ppm；¹³C NMR(CDCl₃, 100MHz) and δ 171.4,151.5,119.0,112.4,62.7,56.1,53.1ppm；IR is (thin Film) v 3371,2953,1741,1685,1548,1529,1475,1357,1306,1216,1076,740cm^-1。

Sequentially added into alcohol JVM.1 (16.3g, 76.8mmol) 110mL DMF solutions imidazoles (6.80g, 99.9mmol, 1.3 equivalents) and t-BuPh₂SiCl (20.6ml, 80.7mmol, 1.05 equivalent).Stirring reaction mixture 14h, is used 300mL Et₂O dilutes, then with 250mL H₂O is quenched.Content is transferred in separatory funnel, organic phase is collected and uses 2x 250mL H₂O and 1x 250mL NaCl saturated aqueous solutions washing.The ether extract magnesium sulfate is dried and is concentrated under reduced pressure. With Silica gel chromatography oily residue (gradient elution：1: 0 → 4: 1 hexane/EtOAc), silyl ether JVM.2 is obtained, For lurid oily (32.1g, 93%).TLC R_f=0.51 (3: 1 hexanes/EtOAc)；¹H NMR(CDCl₃, 400MHz) and δ 7.60-7.57 (m, 4H), 7.46-7.30 (m, 6H), 7.16 (dd, 2H, J=2.4,2.4Hz), 6.41 (d, 1H, J=8.0Hz), 6.29 (dd, 2H, J=2.4,2.4Hz), 4.70 (ddd, 1H, J=8.0,2.4,2.4Hz), 4.20 (dd, 1H, J=10.4, 2.4Hz), (s, the 9H) ppm of 4.01 (dd, 1H, J=10.4,3.2Hz), 3.79 (s, 3H), 1.05；¹³C NMR(CDCl₃, 100MHz) δ 170.5,150.2,135.4,135.3,132.6,132.4,130.00,129.98,127.85,127.82, 118.4,112.1,64.2,55.4,52.7,26.7,19.2ppm；IR (film) v 3361,2954,2858,1748,1711, 1508,1473,1357,1113,736,703cm^-1；HRMS(ES⁺)：C₂₅H₃₀N₂O₄Si calculated values are 450.1975, and measured value is 473.1869(MNa⁺)。

By methyl esters JVM.2 (10.3g, 22.9mmol) 230mL CH₂Cl₂Solution is cooled to -91 DEG C, is added dropwise theretoⁱBu₂AlH (22.9mL 1.50M PhMe solution, 34.3mmol, 1.5 equivalents) 15min.Mixture 4h is stirred at -91 DEG C, Then second part is added¹Bu₂AlH (7.6mL 1.50M PhMe solution, 11.4mmol, 0.5 equivalent).It is stirred for carrying out the reaction Up to 1.5h, 30mL EtOAc is then slowly added at such a temperature is quenched reaction.Content is poured into equipped with 220mL's 1.0M aqueous sodium potassium tartrates and 440mL EtOAc Erlenmeyer (Erlenmeyer) flask, and be stirred vigorously 3.5h.Then content is transferred in separatory funnel.Organic phase is collected, MgSO is used₄Dry and be concentrated under reduced pressure, obtain light yellow Oil.It is not purified and directly using the material so that the loss of optical purity reaches minimum.Pass through silica gel chromatography (9: 1 → 3 : 1 hexane/EtOAc) obtain the sample of pure aldehyde.TLC R_f=0.51 (3: 1 hexanes/EtOAc)；¹H NMR(CDCl₃, 400MHz) and δ 9.72 (s, 1H), 7.63-7.57 (m, 4H), 7.52-7.36 (m, 6H), 7.21 (dd, 2H, J=2.4,2.4Hz), 6.49 (br D, 1H, J=6.4Hz), 6.35 (dd, 2H, J=2.4,2.4Hz), 4.72-4.68 (m, 1H), 4.40 (dd, 1H, J=11.2, 2.8Hz), 4.12 (dd, 1H, J=10.8,3.6Hz), 1.12 (s, 9H) ppm.

By unpurified aldehyde (22.9mmol, it is assumed that yield is 220mL CH 100%)₂Cl₂Solution is cooled to 0 DEG C, Xiang Qi Middle addition allylamine (1.71mL, 22.9mmol).The mixture is heated to room temperature with 20min, -78 DEG C are subsequently cooled to.With 10min is by BF₃·OEt₂(10.2mL, 80.0mmol, 3.5 equivalent) is added dropwise in the solution.Afterwards, content is heated to 23 ℃.Stirring carries out the reaction and reaches 1h, then adds 220mL saturation NaHCO₃The reaction is quenched in the aqueous solution.It is stirred vigorously this Content 20min, is diluted with 440mL EtOAc, is then transferred in separatory funnel.Collect organic phase and with 2x 140mL's EtOAc aqueous layer extracteds.The organic extract magnesium sulfate of merging is dried and is concentrated under reduced pressure.With silica gel chromatography (gradient elution： 4: 1 → 1: 1 hexane/EtOAc) purifying oily residue, amine JVM.3 is obtained, is that (5.9g, two steps are obtained pale yellow foam 56%).TLC R_f=0.21 (1: 1 hexane/EtOAc)；[α]_Na- 93.7 ° of (c=4.11, CDCl₃)；¹H NMR(CDCl₃, 400MHz) δ 7.61-7.55 (m, 4H), 7.43-7.31 (m, 6H), 7.28 (dd, 1H, J=3.2,1.6Hz), 6.17 (dd, 1H, J =3.2,3.2Hz), 6.11-6.09 (m, 1H), 5.88-5.78 (m, 2H), 5.18 (dd, 1H, J=17.2,1.6Hz), 5.10 (dd, 1H, J=10.4,1.6Hz), 3.96 (d, 1H, J=3.2), 3.73 (ddd, 1H, J=10.4,6.8,3.6Hz), 3.60- (s, the 9H) ppm of 3.52 (m, 2H), 3.30-3.17 (m, 2H), 1.02；¹³C NMR(CDCl₃, 100MHz) and δ 149.6,136.2, 135.41,135.38,132.7,132.6,129.80,129.77,128.8,127.73,127.69,117.9,116.5, 110.6,110.4,64.2,58.2,49.13,49.07,26.6,19.0ppm；IR (film) v 3247,2931,2858,1716, 1428,1113,733,703cm^-1；HRMS(ES⁺)：C₂₇H₃₃N₃O₂Si calculated values are 459.2342, and measured value is 482.2235 (MNa⁺)。

(5.61g, 35.9mmol, 3.0 work as mixed amine JVM.3 (5.50g, 12.0mmol), 1,3- dimethyl barbituric acids Amount) and Pd (PPh₃)₄(277mg, 0.24mmol, 0.02 equivalent), and use N₂Flask is purged into a few minutes.Add into reaction vessel Enter 120mL deoxidation CH₂Cl₂(3x freezings/pumping/thaw cycle).Reactant mixture 8h is stirred, order adds 125mL afterwards 1.0M Na₂CO₃The aqueous solution and imines ester JVM.4 (3.84g, 12.0mmol).Stir the two-phase mixture 20min, Ran Houyong 240mL EtOAc dilutions, and be transferred in separatory funnel.Organic phase is collected, and with 1x 120mL EtOAc aqueous layer extracteds.Will The organic extract magnesium sulfate of merging is dried and is concentrated under reduced pressure.With silica gel chromatography (gradient elution：9: 1 → 2: 1 hexane/ EtOAc oily residue) is purified, isothiourea JVM.5 is obtained, is yellow foam (8.1g, 96%).TLC R_f=0.45 (3: 2 Hexane/EtOAc)；[α]_Na- 83.2 ° (c=1.0, MeOH)；¹H NMR(C₆D₆, 400MHz) and δ 8.35 (br d, 1H, J= 8.0Hz), 7.64-7.56 (m, 4H), 7.41 (dd, 1H, J=3.2,1.6Hz), 7.26-7.20 (m, 6H), 6.14 (br s, 1H), 5.95 (dd, 1H, J=3.2,3.2Hz), 5.31 (br s, 1H), 5.02 (br s, 1H), 4.50 (br s, 2H), 3.33- 3.24 (m, 2H), 2.83 (br s, 1H), 1.82 (br s, 3H), 1.09 (s, 9H) ppm；¹³C NMR(CDCl₃, 100MHz) and δ 170.5,148.9,135.50,135.47,132.3,132.1,130.3,130.1,128.0 (2), 124.5,119.3,112.4, 111.6,93.6,78.6,63.1,57.7,47.5,26.8,19.2,14.7ppm；IR (film) v 3287,2931,2858, 1717,1428,1349,1164,1113,738,703cm^-1；HRMS(ES⁺)：C₂₈H₃₃Cl₃N₄O₅S₂Si calculated values are 702.0727, Measured value is 725.0629 (MNa⁺)。

Into the 100mL heavy wall test tubes equipped with magnetic stirring bar add isothiourea JVM.5 (1.95g, 2.76mmol), NH₄OAc (1.1g, 13.8mmol, 5.0 equivalent) and NH₃(27mL 2.0M MeOH solution, 54.0mmol, 19.6 equivalents).With Teflon (Teflon) the spiral cap closure container, and the heating content 24h at 60 DEG C.The reactant mixture is cooled to room Temperature is simultaneously concentrated under reduced pressure.With silica gel chromatography (gradient elution：1: 1 hexane of hexane/EtOAc) purifying, guanidine JVM.6 is obtained, is white Solid (1.64g, 88%).TLC R_f=0.37 (1: 1 hexane/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 7.62-7.56 (m, 4H), 7.42-7.32 (m, 6H), 7.26-7.24 (m, 1H), 6.28-6.26 (m, 1H), 6.21 (dd, 1H, J=3.2,3.2Hz), 5.46 (br s, 1H), 4.62 (s, 2H), 3.80-3.76 (m, 1H), 3.66 (dd, 1H, J=10.4,4.8Hz), 3.45 (dd, 1H, J=10.0,7.6Hz), 0.99 (s, 9H) ppm；¹³C NMR(CD₃OD, 100MHz) δ 156.5,150.2,135.6,135.5, 132.7,132.5,129.90,129.87,127.83,127.81,127.2,117.9,111.19,111.14,94.5,78.2, 64.3,57.4,43.6,26.2,18.9ppm；IR (film) v 3355,2932,2859,2455,1695,1544,1445, 1298,1114,1019,848,739cm^-1。

Tces guanidines JVM.6 (422mg, 0.63mmol), Rh are added into 100mL round-bottomed flasks₂(esp)₂(33mg, 44 μ Mol, 0.07 equivalent), PhI (OAc)₂(404mg, 1.25mmol, 2.0 equivalent) and MgO (114mg, 2.8mmol, 4.4 equivalent). 25mL toluene is added into the solid of merging.The bottle green reactant mixture of acquisition is heated into 3h between 40-45 DEG C.Then Mixture is cooled to room temperature and direct silicagel column excessively.Gradient elution (hexane → 1: 1 hexane/EtOAc), obtains guanidine JVM.7, is White solid (200mg, 43%).TLC R_f=0.41 (1: 1 hexane/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 7.63- 7.60 (m, 4H) .7.47-7.37 (m, 6H), 6.44 (d, 1H, J=2.0Hz), 6.25 (d, 1H, J=5.6Hz), 6.07 (dd, 1H, J=5.6,2.0Hz), 4.63 (s, 1H), 4.62 (s, 1H), 4.46 (d, 1H, J=1.4Hz), 3.65-3.60 (m, 1H), (s, the 9H) ppm of 3.48 (dd, 1H, J=3.0,6.0Hz), 2.05 (s, 3H), 1.05；¹³C NMR(CDCl₃, 125MHz) and δ 170.5, 157.9,154.3,135.79,135.77,133.4,132.3,132.2,130.6,130.5,130.4,128.36,128.33, 94.1,85.8,83.6,78.4,63.9,56.4,55.7,27.1,21.5,19.4ppm；IR (film) v 3345,2932, 2859,1685,1612,1428,1114,734,703cm^-1。

By allyl acetate JVM.7 (171mg, 0.23mmol) and triethyl silicane (188 μ L, 1.17mmol, 5.0 equivalent) In 5.0mL CH₂Cl₂In solution be cooled to -78 DEG C, and add BF₃·OEt₂(68 μ L, 0.54mmol, 2.3 equivalent).Make anti- Answer content to be to slowly warm up to room temperature, and stir 3h.Then 5mL NaHCO is added₃The reaction is quenched in saturated aqueous solution, and It is stirred vigorously mixture 15min.The content is diluted with 10mL EtOAc and is transferred in separatory funnel.Collected organic layer And with 2x 10mL EtOAc aqueous layer extracteds.The organic extract magnesium sulfate of merging is dried and is concentrated under reduced pressure.Use silica gel color Spectrometry (gradient elution：Hexane → 2: 1 hexane/EtOAc) material that is isolated and purified with, alkene JVM.8 is obtained, is white solid (121mg, 77%).TLC R_f=0.22 (1: 1 hexane/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 7.64-7.60 (m, 4H), 7.46-7.38 (m, 6H), 6.15-6.12 (m, 1H), 5.92-5.89 (m, 1H), 4.60 (s, 2H), 4.43 (d, 1H, J= 1.2Hz), 4.13 (ddd, 17.0,2.2,2.2Hz, 3.81 (ddd, 1H, J=17.0,2.2,2.2Hz), 3.59-3.51 (m, 1H), 3.45 (dd, 1H, J=7.2,9.6Hz), 1.04 (s, 9H) ppm；¹³C NMR(CDCl₃, 125MHz) and δ 157.8,156.2, 135.82,135.77,132.5,132.4,131.3,130.5,130.4,128.3,128.2,94.1,84.3,78.4,64.0, 57.8,57.3,52.2, (30.0), 27.1,19.4ppm；IR (film) v 3231,2930,1673,1631,1526,1183, 1114,732,702cm^-1。

To isothiourea JVM.5 (6.62g, 9.4mmol) and 2, (8.64g, 34.8mmol, 3.7 work as 4,6- tri-tert pyrimidines Amount) in 19.0mL CH₂Cl₂Solution in add trifluoromethanesulfonic acid ethyl ester (7.3mL, 56.3mmol, 6.0 equivalent).Use glass stopper Closed reaction vessel, heats the solution to 37 DEG C and stirs 14h.Then, cooling reaction system adds 50mL's to room temperature CH₂Cl₂Dilution, and it is transferred to NaHCO containing 570mL₃In Erlenmeyer (Erlenmeyer) flask of saturated aqueous solution.Acutely stir Two-phase mixture 12h is mixed, is diluted with 150mL EtOAc, and be transferred in separatory funnel.Collected organic layer simultaneously uses 2x 150mL EtOAc aqueous layer extracteds.The organic extract magnesium sulfate of merging is dried and is concentrated under reduced pressure.With silica gel chromatography (ladder Degree elution：1: 0 → 3: 1 hexane/EtOAc) purifying oily residue, obtain isourea JVM.9, be yellow foam (5.30g, 77%).TLC R_f=0.34 (3: 1 hexanes/EtOAc)；¹H NMR(CDCl₃, 400MHz) and δ 8.08 (d, 1H, J=8.8Hz), 7.65-7.58 (m, 4H), 7.43-7.30 (m, 6H), 7.03 (dd, 1H, J=3.2,1.6Hz), 6.35 (dd, 1H, J=3.2, 0.8Hz), 6.19 (dd, 1H, J=3.2,3.2Hz), 5.38 (dd, 1H, J=8.6,3.4Hz), 4.62 (s, 2H), 4.36-4.19 (m, 2H), 4.02 (ddd, 1H, J=7.6,3.8,3.8Hz), 3.75 (dd, 1H, J=10.2,4.0Hz), 3.25 (dd, 1H, J= 10.2,8.2Hz) (s, the 9H) ppm of, 2.50 (s, 3H), 1.35 (t, 3H, J=7.0Hz), 1.04；¹³C NMR(CDCl₃, 100MHz) δ 170.6,146.7,135.5,135.4,132.8,132.7,129.8,129.7,127.7,127.6,125.2,117.0, 111.0,110.4,93.6,78.4,63.4,62.8,60.8,47.7,26.6,19.1,14.5,14.0ppm；IR (film) v 3289,2932,2858,1669,1561,1428,1333,1160,1113,732cm^-1；HRMS(ES⁺)：C₃₀H₃₇Cl₃N₄O₅S₂Si Calculated value is 730.1040, and measured value is 753.0940 (MNa⁺)。

Isourea JVM.9 (1.98g, 2.71mmol), NH are added into the 100mL heavy wall test tubes equipped with magnetic stirring bar₄OAc (1.0g, 13.0mmol, 4.8 equivalent) and NH₃(40mL 2.0M MeOH solution, 80.0mmol, 29.5 equivalents).Use Teflon The spiral cap closure container, heats content 24h at 70 DEG C.The reactant mixture is cooled to room temperature and is concentrated under reduced pressure. Non- purified product is carried out¹H NMR analysis shows, the purity of gained material is sufficient for subsequently using.

Unpurified guanidine is dissolved in 13.0mL MeCN, and addedⁱPr₂NEt (2.36mL, 13.5mmol, 5.0 equivalent).It is cold But the solution is to 0 DEG C, and adds pure CF₃CO₂Et (966 μ L, 8.12mmol, 3.0 equivalent).Content is to room temperature and stirs for heating Mix 24h.Then extra number is addedⁱPr₂NEt (2.36mL, 13.5mmol, 5.0 equivalent) and CF₃CO₂Et (966 μ L, 8.12mmol, 3 equivalents).Stir after mixture 24h, decompression removes all volatile matters.With silica gel chromatography (gradient elution：Oneself Alkane → 2: 1 hexane/EtOAc) product that is isolated and purified with, biguanides JVM.10 is obtained, is white solid (1.64g, 79%).TLC R_f=0.43 (2: 1 hexanes/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 7.58-7.52 (m, 5H), 7.42-7.23 (m, 6H), 6.40-6.38 (m, 1H), 6.30 (dd, 1H, J=3.2,3.2Hz), 5.41 (br s, 1H), 4.64 (d, 1H, J=11.2Hz), 4.60 (d, 1H, J=11.2Hz), 4.07 (ddd, 1H, J=5.2,5.2,2.4Hz), 3.78-3.66 (m, 2H), 0.94 (s, 9H) ppm；¹³C NMR(CDCl₃, 100MHz) and δ 169.2 (q, J=36.4Hz), 155.9,154.6,135.54,135.50,132.13, 132.10,130.3 (2H), 128.1 (2H), 125.0,120.1,116.4 (d, J=284Hz), 114.5,113.2,94.0, 78.3,63.5,57.2,43.7,26.8,19.1ppm；IR (film) v 3448,3351,1645,1535,1261,1202, 1116,855,736,702cm^-1。

Tces guanidines JVM.10 (131mg, 0.17mmol), Rh are added into 50mL round-bottomed flasks₂(esp)₂(6.5mg, 8.6 μ Mol, 0.05 equivalent), PhI (OAc)₂(137mg, 0.43mmol, 2.5 equivalent) and MgO (31mg, 0.77mmol, 4.5 equivalent). 6.8mL toluene is added into the solid of merging.The bottle green reactant mixture that acquisition is heated at 42 DEG C reaches 3h.Then, will The mixture is cooled to room temperature, and directly crosses silicagel column.Gradient elution (hexane → 1: 1 hexane/EtOAc), obtains guanidine JVM.11, is white solid (74mg, 52%).TLC R_f=0.39 (3: 2 hexanes/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 7.63-7.56 (m, 4H), 7.48-7.39 (m, 6H), 6.80 (dd, 1H, J=2.0,0.8Hz), 6.31 (dd, 1H, J=6.0, 0.4Hz), 6.09 (dd, 1H, J=6.0,2.0Hz), 4.65 (d, 1H, J=11.2Hz), 4.62 (d, 1H, J=11.2Hz), 4.33 (d, 1H, J=2.8Hz), 3.86-3.82 (m, 1H), 3.74 (dd, 1H, J=10.4,4.4Hz), 3.51 (dd, 1H, J= 7.6,10.4Hz), 2.07 (s, 3H), 1.01 (s, 9H)；¹³C NMR(CDCl₃, 125MHz) and δ 169.8,168.1 (q, J= 36.5Hz), 159.1,157.7,135.55,135.52,132.5,131.9,131.7,130.6,130.5,130.4,128.2, 128.1,116.6 (d, J=285.0Hz), 93.7,84.7,81.4,78.2,63.4,57.0,55.8,26.7,20.9, 19.1ppm；IR (film) v 3282,2933,2860,1743,1622,1429,1351,1150,1017,853,703cm^-1。

By allyl acetate JVM.11 (352mg, 0.43mmol) and triethyl silicane, (688 μ L, 4.26mmol, 10.0 work as Amount) in 9.0mL CH₂Cl₂In solution be cooled to -78 DEG C, and add BF₃·OEt₂(216 μ L, 1.70mmol, 4.0 equivalent). Reaction content is to slowly warm up to 0 DEG C and stir 3h at such a temperature.Then, 15mL NaHCO is added₃Saturated aqueous solution makes The reaction is quenched.It is stirred vigorously the two phase liquid 15min of acquisition.Then the mixture is diluted with 25mL EtOAc, and shifted Into separatory funnel.Collected organic layer and with 2x 25mL EtOAc aqueous phase extracteds.By the organic extract magnesium sulfate of merging Dry and be concentrated under reduced pressure.With silica gel chromatography (gradient elution：Hexane → 2: 1 hexane/EtOAc) material that is isolated and purified with, obtain It is white solid (251mg, 73%) to alkene JVM.12.TLC R_f=0.45 (1.5: 1 hexanes/EtOAc)；)；¹H NMR (CD₃OD, 400MHz) δ 7.64-7.57 (m, 4H), 7.48-7.37 (m, 6H), 6.17 (ddd, 1H, J=6.0,2.0,2.0Hz), 5.93 (ddd, 1H, J=6.0,2.0,2.0Hz), 4.62 (d, 1H, J=11.2Hz), 4.59 (d, 1H, J=11.2Hz), 4.35 (m, 2H), 4.09 (ddd, 17.0,2.2,2.2Hz, 3.81-3.77 (m, 1H), 3.71 (dd, 1H, J=11.2,5.2Hz), 3.54 (dd, 1H, J=11.2,8.4Hz), 1.01 (s, 9H) ppm；¹³C NMR(CDCl₃, 125MHz) and δ 167.8 (q, J=35.6Hz), 160.2,157.8,135.79,135.77,132.2,132.1,131.9,130.50,130.48,128.3 (2H), 127.1, 116.8 (d, J=285Hz), 93.9,82.4,78.4,63.7,59.1,56.1,53.9,26.9,19.3ppm；IR (film) v 3286,2933,1634,1569,1429,1114,907,736cm^-1。

Isourea JVM.9 (3.98g, 5.44mmol), NH are added into the 100mL heavy wall test tubes equipped with magnetic stirring bar₄OAc (2.10g, 27.2mmol, 5.0 equivalent) and NH₃(32mL 2.0M MeOH solution, 64.0mmol, 11.8 equivalents).Use Teflon The spiral cap closure container, and heat at 70 DEG C content 24h.The reactant mixture is cooled to room temperature and depressurized dense Contracting.With silica gel chromatography (gradient elution：1: 1 hexane/EtOAc → 3: 17MeOH/EtOAc) material that is isolated and purified with, obtain Biguanides JVM.13, is white foam (3.38g, 85%).TLC R_f=0.31 (6: 1 EtOAc/MeOH)；¹H NMR(CD₃OD, 500MHz) δ 7.56-7.61 (m, 4H), 7.36-7.45 (m, 7H), 6.45 (d, 1H, J=3.4Hz), 6.42 (dd, 1H, J= 3.4,3.4Hz), 5.44 (bs, 1H), 4.61 (d, 1H, J=2.5Hz), 3.91-3.95 (m, 1H), 3.70 (dd, 1H, J= 10.8,4.5Hz) (s, the 9H) ppm of, 3.60 (dd, 1H, J=10.6,5.8Hz), 1.91 (s, 3H), 0.96；¹³C NMR(CD₃OD, 125MHz) δ 157.7,151.9,136.73,136.72,133.59,133.47,131.2 (2H), 129.09,129.08, 128.5,119.2,115.5,114.2,95.7,79.3,65.3,57.6,44.8,27.4,20.0ppm；IR (film) v 2932, 1699,1627,1542,1427,1330,1184,1114,1019,846,703cm^-1。

By biguanides JVM.13 (3.24g, 4.43mmol) 44mL CH₂Cl₂Solution is cooled to -20 DEG C, and order adds thereto EnterⁱPr₂NEt (1.54mL, 8.84mmol, 2.0 equivalent) and trichloro-acetic chloride (572 μ L, 5.10mmol, 1.15 equivalent).- 20 Reactant mixture 1h is stirred at DEG C, 50mL NaHCO is then added₃The reaction is quenched in saturated aqueous solution.With 100mL's EtOAc dilutes the two-phase content, and is transferred in separatory funnel.Collected organic layer and with 2x 50mL EtOAc extraction waters Phase.The organic extract magnesium sulfate of merging is dried and is concentrated under reduced pressure.With silica gel chromatography (gradient elution：1: 0 → 3: 1 oneself Alkane/EtOAc) material that is isolated and purified with, guanidine JVM.14 is obtained, is white solid (3.14g, 89%).TLC R_f=0.43 (2: 1 hexane/EtOAc)；mp 174-176℃；[α]_Na- 51.6 ° (c=0.71, MeOH)；¹H NMR(CD₃OD, 400MHz) δ 7.59 (dd, 1H, J=3.2,1.6Hz), 7.58-7.53 (m, 4H), 7.40-7.32 (m, 6H), 6.40-6.37 (m, 1H), 6.28 (dd, 1H, J=3.2,3.2Hz), 5.42 (br s, 1H), 4.65 (d, 1H, J=11.2Hz), 4.61 (d, 1H, J=11.2Hz), 4.12 (ddd, 1H, J=5.2,5.2,2.4), 3.75-3.66 (m, 2H), 0.93 (s, 9H) ppm；¹³C NMR(CD₃OD, 125MHz) δ 173.9,157.5,155.5,136.57,136.55,133.4,133.3,131.0 (2), 128.96,128.92,127.3, 120.5,114.0,113.4,97.3,95.5,79.2,65.4,58.2,44.9,27.2,19.8ppm；IR (film) v 3464, 3358,2931,2859,1603,1589,1363,1212,1115,840,739cm^-1；HRMS(ES⁺)：C₂₉H₃₂Cl₆N₆O₅SSi is counted Calculation value is 814.0055, and measured value is 836.9949 (MNa⁺)。

Tces guanidines JVM.14 (1.675g, 2.05mmol), Rh are added into 200mL round-bottomed flasks₂(esp)₂(78mg, 0.10mmol, 0.05 equivalent), PhI (OAc)₂(1.65g, 5.12mmol, 2.5 equivalent) and MgO (372mg, 9.23mmol, 4.5 Equivalent).81mL CH is added into the solid of merging₂Cl₂.The flask is closed with glass stopper, the dark green of acquisition is heated at 42 DEG C Colour response mixture reaches 2.5h.Then, the suspension is filtered by a fritter Celite pad.Use CH₂Cl₂Rinse the flask and filter Cake.80mL 1: 1 NaHCO is added into the filtrate of merging₃Saturated aqueous solution/saturation Na₂S₂O₃Solution.The mixture is turned Separatory funnel, collected organic layer are moved to, and aqueous phase is extracted with 2x 50mL EtOAc.By the organic extract magnesium sulfate of merging Dry and be concentrated under reduced pressure.With silica gel chromatography (gradient elution：1: 0 → 2: 1 hexane/EtOAc) purifying oily residue, obtain Three required ring JVM.15, are white solid (1.12g, 62%).TLC R_f=0.27 (2: 1 hexanes/EtOAc)；¹H NMR (CD₃OD, 400MHz) δ 7.64-7.58 (m, 4H), 7.47-7.40 (m, 6H), 6.86 (d, 1H, J=2.0Hz), 6.32 (d, 1H, J=6.0Hz), 6.10 (dd, 1H, J=5.8,2.2Hz), 4.67 (d, 1H, J=11.2Hz), 4.63 (d, 1H, J=11.0Hz), 4.36 (d, 1H, J=2.4Hz), 3.86 (ddd, 1H, J=8.0,4.4,2.4Hz), 3.73 (dd, 1H, J=10.4,4.4Hz), (s, the 9H) ppm of 3.51 (dd, 1H, J=10.6,7.8Hz), 2.09 (s, 3H), 1.02；¹³C NMR(CDCl₃, 125MHz) and δ 172.6,169.6,158.8,157.4,135.52,135.50,132.4,131.8,131.7,130.6,130.4,130.3, 128.17,128.13,95.8,93.7,85.2,81.4,78.2,63.6,56.8,55.7,26.8,21.1,19.1ppm；IR is (thin Film) v 3286,2932,2859,1742,1624,1574,1374,1179,1088,1015,909,831,735,703cm^-1； HRMS(ES⁺)：C₃₁H₃₄Cl₆N₆O₇SSi calculated values are 872.0110, and measured value is 895.0004 (MNa⁺)。

By allyl acetate JVM.15 (1.02g, 1.16mmol) and triethyl silicane, (940 μ L, 5.82mmol, 5.0 work as Amount) in 24.0mL CH₂Cl₂In solution be cooled to -78 DEG C, and add BF₃·OEt₂(339 μ L, 2.68mmol, 2.3 equivalent). Reaction content is to slowly warm up to room temperature and stir 1.5h.Then, 40mL NaHCO is added₃Saturated aqueous solution makes the reaction It is quenched.The two phase liquid for being stirred vigorously acquisition reaches 15min.The mixture is diluted with 50mL EtOAc, and is transferred to a point liquid leakage In bucket.Collected organic layer and with 2x 25mL EtOAc aqueous layer extracteds.The organic extract magnesium sulfate of merging is dried and subtracted Pressure concentration.With silica gel chromatography (gradient elution：Hexane → 2: 1 hexane/EtOAc) material that is isolated and purified with, obtain alkene JVM.16, is white solid (792mg, 83%).TLC R_f=0.48 (2: 1 hexanes/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 7.65-7.59 (m, 4H), 7.46-7.37 (m, 6H), 6.17 (ddd, 1H, J=6.0,1.8,1.8Hz), 5.95 (ddd, 1H, J=6.0,2.2,2.2Hz), 4.63 (d, 1H, J=11.2Hz), 4.60 (d, 1H, J=11.2Hz), 4.37 (ddd, 1H, J=17.0,2.2,2.2Hz), 4.32 (d, 1H, J=2.4Hz), 4.17 (ddd, 1H, J=17.0,2.0,2.0Hz), 3.82 (ddd, 1H, J=8.0,4.4,2.0Hz), 3.70 (dd, 1H, J=10.6,4.8Hz), 3.55 (dd, 1H, J=10.6, 8.0Hz), 1.02 (s, 9H) ppm；¹³C NMR(CDCl₃, 125MHz) and δ 172.4,159.7,157.7,135.6 (2), 131.9 (2), 131.7,130.3 (2), 128.1 (2), 127.0,96.3,93.8,82.3,78.2,63.8,58.3,55.8,53.8, 26.8,19.1ppm；IR (film) v 3281,2933,2859,1631,1565,1377,1179,1113,908,840,734cm^-1；HRMS(ES⁺)：C₂₉H₃₂Cl₆N₆O₅SSi calculated values are 814.0055, and measured value is 836.9951 (MNa⁺)。

Alkene JVM.16 (206mg, 0.25mmol) 5.0mL THF solutions are cooled to -78 DEG C, and add tetrabutyl fluorine Change ammonium (305 μ L 1.0M THF solutions, 0.305mmol, 1.2 equivalents).The mixture is heated to 0 DEG C and stirred at such a temperature Mix 20min.Then, 5.0mL NH is added₄The reaction is quenched in Cl saturated aqueous solutions.The content is diluted with 10mL EtOAc Thing, and be transferred in separatory funnel.Collected organic layer and with 3x 10mL EtOAc aqueous phase extracteds.By the organic extract of merging Dried and be concentrated under reduced pressure with magnesium sulfate.¹The purity of the H NMR analysis shows material is suitable, directly can be used in subsequent reactions. Pure desilylation alcohol sample is obtained with silica gel chromatography (2: 1 → 1: 2 hexanes/EtOAc).TLC R_f=0.33 (1: 2 oneself Alkane/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 6.28 (ddd, 1H, J=6.2,2.0,2.0Hz), 6.08 (ddd, 1H, J= 6.2,2.2,2.2Hz), 4.67 (d, 1H, J=11.2Hz), 4.64 (d, 1H, J=11.2Hz), 4.43 (m, 2H), 4.35 (d, 1H, J=2.8Hz), 3.69-3.63 (m, 2H), 3.57 (dd, 1H, J=12.4,8.4Hz) ppm；¹³C NMR(CDCl₃, 125MHz) δ 172.3,159.4,157.9,130.9,127.4,96.5,94.1,83.1,78.4,61.7,55.5,55.2, 54.5ppm；IR (film) v 3289,1616,1562,1379,1175,905,841,730cm^-1。

By the 10.0mL CH of unpurified alcohol₂Cl₂Solution is cooled to -20 DEG C, and trichloroacetyl isocyanate (508 μ are added dropwise L 0.5M CH₂Cl₂Solution, 0.25mmol).The mixture is stirred into 15min at -20 DEG C, diluted with 20mL EtOAc, and Add 10mL NaHCO₃The reaction is quenched in saturated aqueous solution.Content is transferred to separatory funnel and collected organic layer.With 2x 10mL EtOAc aqueous layer extracteds.The organic extract of merging is dried with magnesium sulfate, filtered and is concentrated under reduced pressure.It will isolate Material be re-dissolved in 10.0mL MeOH and stir solution 12h.Then, the mixture is concentrated under reduced pressure.Use silica gel chromatograph Method (gradient elution：Hexane → 1: 2 hexanes/EtOAc) purifying oily residue, carbamate JVM.17 is obtained, is that white is solid (76%) 121mg, two steps obtain body.TLC R_f=0.18 (1: 2 hexane/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 6.32 (ddd, 1H, J=6.2,2.0,2.0Hz), 6.04 (ddd, 1H, J=6.2,2.2,2.2Hz), 4.64 (d, 1H, J=11.2Hz), 4.61 (d, 1H, J=11.2Hz), 4.44 (d, 1H, J=2.4Hz), 4.43-4.40 (m, 2H), 4.15-4.03 (m, 2H), 3.86 (ddd, 1H, J=7.4,4.8,2.4Hz) ppm；¹³C NMR(CD₃OD, 125MHz) δ 172.9,161.3,159.0,158.6, 131.7,129.0,97.7,95.5,83.3,79.4,64.3,60.5,55.0,54.7ppm；IR (film) v 3292,2951, 2457,1723,1617,1565,1494,1341,1175,1086,895,842,752cm^-1；HRMS(ES⁺)：C₁₄H₁₅Cl₆N₇O₆S Calculated value is 618.8936, and measured value is 641.8830 (MNa⁺)。

N-methylmorpholine-N- oxygen is sequentially added into alkene JVM.17 (117mg, 0.20mmol) 4.0mL THF solutions Compound (44mg, 0.38mmol, 2.0 equivalent) and OsO₄(24 μ L 4% aqueous solution, 3.8 μm of ol, 0.02 equivalent).Stir this anti- Mixture 12h is answered, 4mL Na is then added₂S₂O₃Reaction is quenched in saturated aqueous solution.The content is diluted with 8mL EtOAc, And be transferred in separatory funnel.Collected organic layer and with 2x 4mL EtOAc aqueous phase extracteds.By the organic extract sulphur of merging Sour magnesium is dried and is concentrated under reduced pressure.With silica gel chromatography (gradient elution：2: 1 → 0: 1 hexane/EtOAc) purifying oily residue, Glycol JVM.18 is obtained, is white solid (102mg, 82%).TLC R_f=0.33 (neat EtOAc)；¹H NMR (CD₃OD.400MHz) δ 4.63 (s, 2H), 4.56 (d, 1H, J=2.8Hz), 4.54 (ddd, 1H, J=7.6,7.6,4.0Hz), 4.21-4.14 (m, 2H), 3.98 (dd, 1H, J=11.6,8.0Hz), 3.95 (d, 1H, J=4.0Hz), 3.79 (ddd, 1H, J= 7.2,6.0,2.8Hz), 3.45 (dd, 1H, J=11.6,7.6Hz) ppm；¹³C NMR(CD₃OD, 125MHz) δ 171.6,160.4, 158.2,157.8,96.7,94.3,80.7,78.2,76.1,67.9,63.0,55.7,53.4,50.0ppm；IR (film) v 3299,2985,1711,1618,1567,1376,1321,1177,1088,902,845,760cm^-1；HRMS(ES⁺)： C₁₄H₁₇Cl₆N₇O₈S calculated values are 652.8990, and measured value is 675.8892 (MNa⁺)。

By benzoyl cyanide (1.64mL 0.1M CH₂Cl₂Solution, 0.164mmol, 1.1 equivalents) it is added drop-wise to -78 DEG C of glycol JVM.18 (98mg, 0.149mmol) and 4-dimethylaminopyridine (72.8mg, 0.60mmol, 4.0 equivalent) are in 6.0mL 3: 1 CH₂Cl₂In solution in/MeCN mixtures.Stirring carries out the reaction up to 1.5h at -78 DEG C, is then quenched with 50 μ L MeOH Go out.With 10mL EtOAc and 5mL NaHCO₃Saturated aqueous solution dilutes the content, and is transferred in separatory funnel.Collection has Machine layer and with 2x 10mL EtOAc aqueous phase extracteds.The organic extract magnesium sulfate of merging is dried and is concentrated under reduced pressure.Use silicon Glue chromatography (gradient elution：Hexane → 1: 2 hexanes/EtOAc) purifying oily residue, benzoic ether JVM.19 is obtained, is white Color solid (78mg, 69%).¹H NMR(CD₃OD, 400MHz) δ 8.12-8.09 (m, 2H), 7.66-7.61 (m, 1H), 7.53- 7.48 (m, 2H), 5.61 (ddd, 1H, J=8.4,7.6,4.0Hz), 4.67 (d, 1H, J=10.8), 4.64 (d, 1H, J= 10.8Hz), 4.63 (d, 1H, J=3.2Hz), 4.40 (d, 1H, J=4.0Hz), 4.29-4.15 (m, 3H), 3.85 (ddd, 1H, J =8.4,5.2,2.8Hz), 3.77 (dd, 1H, J=11.8,7.4Hz) ppm；¹³C NMR((CD₃)₂CO, 100MHz) δ 171.0, 165.5,160.0,158.1,156.15,133.5,129.8,129.5,128.6,94.4,81.0,77.9,74.3,71.0, 62.5,54.4,53.1,47.9ppm；IR (film) v 3294,2985,1715,1620,1274,1177,902,846,760, 716cm^-1。

To benzoic ether JVM.19 (12.7mg, 16.7 μm of ol) 700 μ L CH₂Cl₂Dai Si-Martin's oxidation is added in solution Agent (10.9mg, 26.0 μm of ol, 1.5 equivalents).Stirring carries out the reaction and reaches 20min, then adds 700 μ L 1: 1 NaHCO₃It is full With the aqueous solution/Na₂S₂O₃Reaction is quenched in saturated aqueous solution.The two-phase mixture is stirred vigorously up to 10min, with 1mL CH₂Cl₂ Dilution, and be transferred in separatory funnel.Collected organic layer and with 1x 1mL CH₂Cl₂Aqueous phase extracted.By organic extraction of merging Thing directly crosses silicagel column.With silica gel chromatography (gradient elution：Hexane → 1: 2 hexanes/EtOAc) purifying, JVM.20 is obtained, is White solid (10mg, 78%).¹H NMR((CD₃)₂CO, 400MHz) δ 9.44 (br s, 1H), 8.20-8.17 (m, 2H), 7.70-7.65 (m, 1H), 7.56-7.50 (m, 2H), 6.76 (br s, 1H), 6.65 (br s, 1H), 6.00 (br s, 2H), 5.72 (dd, 1H, J=8.6,7.0Hz), 4.87 (d, 1H, J=2.4Hz), 4.61 (s, 2H), 4.35 (dd, 1H, J=12.2, 8.6Hz), (dd, 1H, J=12.2, the 7.2Hz) ppm of 4.28-4.25 (m, 2H), 4.08 (m, 1H), 3.62；¹³C NMR((CD₃)₂CO, 100MHz) δ 171.0,165.9,160.2,158.6,156.2,133.7,130.0,129.2,128.5,98.4,96.8, 94.4,80.6,77.9,73.6,62.6,54.9,53.7,47.5ppm；IR (film) v 3286,1714,1618,1566, 1318,1274,1175,1088,1016,903cm^-1。

50 μ L CF is added into glycol JVM.20 (5.8mg, 7.6 μm of ol) 2.0mL MeOH solution₃CO₂H.Stir into The row reaction reaches 30min, then adds Pd/C (22mg 10wt.%, 0.021mmol, 2.75 equivalent).Reaction vessel is put in In high pressure Paar (Parr) basin, it is sealed against and uses H₂Gas (800psi) is rinsed six times.The basin is forced into 800psi's H₂, content 12h is stirred at this pressure.Then the basin is vented, with 0.2 μm of PTFE syringe of Fei Xieer (Fisher) Formula filter filters reactant mixture.Flask and filter are washed with 2mL MeOH, and filtrate decompression is concentrated.Film is residual Excess is dissolved in 1.0mL NH₃2.0M MeOH solution in.Stir after solution 15min, decompression removes all volatile materials. The material isolated is directly dissolved in 2.0mL 0.5MCF₃CO₂The H aqueous solution simultaneously stirs 24h.Be concentrated under reduced pressure the solution, obtains thin Membranaceous product, with reverse HPLC-purified (Alltima C18,10 μM, 10x 250mm posts, 1: 99 → 30: 70 MeCN/10mM C₃F₇CO₂The H aqueous solution gradient elution 20min, 214nm UV are detected).Flow velocity is 6mL/min, and 11 β-OH-STX retention time is 13.7-15.5min, isolated white hygroscopic solid (4.6mg, 83%).¹H NMR(D₂O, 400MHz) δ 4.80 (1H), 4.45 (dd, 1H J=8.0,7.2Hz), 4.33 (dd, 1H, J=11.6,9.6Hz), 4.07-3.98 (m, 2H), 3.82 (dd, 1H, J=9.6,4.4Hz), 3.27 (dd, 1H, J=10.4,7.2Hz) ppm；¹³C NMR(D₂O, 600MHz, HMBC and HSQC are true It is fixed) δ 158.7,157.6,155.5,97.6,81.7,70.5,63.0,57.5,53.0,48.5ppm；HRMS(ES⁺)： C₁₀H₁₇N₇O₅Calculated value is 315.1291, and measured value is 316.1366 (MH⁺)。

To 11 β-OH-STX (7.3mg, 9.8 μm of ol) and 2,6- di-t-butyl -4- picolines (55mg, 0.27mmol, 27.5 equivalents) solution in DMFSO is added dropwise₃(9.1 work as by 893 μ L 0.1M 1-METHYLPYRROLIDONE solution, 89.3 μm of ol Amount).Reactant mixture 3h is stirred, 300 μ L H is then added₂Reaction is quenched in O.It is concentrated in vacuo the solution, and with anti-phase HPLC purifies film residue (the Alltima C18 posts, 10 μM, 10x 250mm posts, 1: 99 MeCN/10mM C₃F₇CO₂H water Solution gradient elutes 20min, 214nm UV detections).Flow velocity is 6mL/min, and GTX 3 retention time is 6.8-7.9min, point From obtaining white hygroscopic solid (4.2mg, 71%).[α]_Na+ 43.3 ° of (c=0.40 (CF₃CF₂CF₂CO₂ ^-Salt)；¹H NMR (D₂O, 400MHz) δ 4.96 (dd, 1H, J=8.2,6.8Hz), 4.82 (d, 1H, J=1.2Hz), 4.30 (dd, 1H, J=11.8, 9.4Hz), 4.17 (dd, 1H, J=10.6,8.2Hz), 4.06 (dd, 1H, J=11.8,5.4Hz), 3.82 (ddd, 1H, J= 9.0,5.4,1.2Hz), 3.58 (dd, 1H, J=10.6,6.8Hz) ppm；¹³C NMR(D₂O, 600MHz, it is true by HMBC and HSQC It is fixed) δ 158.7,157.6,155.5,97.3,81.6,75.7,63.0,57.2,53.1,47.4ppm；HRMS(ES⁺)： C₁₀H₁₅N₇O₇S calculated values are 377.0754, and measured value is 378.0833 (MH⁺)。

Synthesize and natural (+)-GTX-3 is (referring to document：Onodera, H.；Satake, M.；Oshima, Y.；Yasumoto, T.；Carmichael, W.W.Nat.Toxins.1997,5,146-151) spectroscopic data contrast

Embodiment 3：

Obtained in the oxazolidone intermediate using preceding face derived from many saxitoxins with linear long-chain branch After molecular probe, we have further synthesized many side chain derivatives using the chemical property.These molecules are in Saxidomus poison There is larger spatial volume near plain parent nucleus, this potentially makes the combination of toxin unstable.It is contemplated that using these molecules simultaneously With reference to mutation sodium channel, so as to draw out the steric environment when STX is attached in the passage around its side chain.In addition, can Saxitoxin derivative is designed to expect introducing extra chemical moieties (" salient point " (bump)), makes it to wild type Passage has low binding affinity and has high binding affinity to the mutant with corresponding " hole ".(referring to document： Annual Review of Cell and Developmental Biology 2001,17,405-433, pass through the side of reference Its content is incorporated herein by formula).

In initial research, we devise one group of molecule, and it is near the nitrogen and the nitrogen of carbamic acid ester side chain Carbon on have side chain.Two mutant channels are built by rite-directed mutagenesis.Initial diaphragm is shown in the table 1 of embodiment 3 Clamp result.β-STX phenol (β-STXol) data point is collected, STX is indicated and its derivative is similarly oriented in access opening.STX β-STX phenol and cyclohexyl STX are similarly reduced to the affinity of cyclohexyl β-STX phenol, are two kinds of molecules all with similar Oriented approach be located at access opening in provide strong primary evidence.

Although these saxitoxin derivatives are not all shown to one of described two mutant compares WT channel Higher affinity, but compared with natural STX, they show significant affinity difference really.Especially when for When M1240A mutant is tested, the unstability degree of parent compound is up to about 40 times, and the unstability journey of cyclohexylamine derivant Degree is only 4 times.This species diversity may imply that methionine sports less alanine, be that carbamate occupies offer Bigger space, so as to preferably accommodate hexamethylene base side chain.The other mutation of sodium channel is being investigated further to study " hole " is created to accommodate the possibility of larger carbamic acid ester side chain (" salient point ").

The table 1 of embodiment 3：Various STX derivatives are to Na_v1.4 and its affinity of mutant

Except the research of these configuration aspects, we have also been devised other probes for being imaged in vivo to sodium channel. Specifically, we devise such STX derivatives, wherein can be introduced by the stable oxime key of physiology¹⁸F- marks Benzaldehyde.(referring to document：Journal of Nuclear Medicine 2008,49,804-813, by reference will Its content is incorporated herein.) intermediate of hexylamine end-blocking that generally uses from us synthesizes the exemplary route of synthesis of the molecule As shown in figure 12.Figure 12 shows the synthetic schemes of synthesis PET image probes.Study on the synthesis shows, can successfully carry out Initial coupling, and Cheng et al. work confirms oxime key and can be readily formed and be stable in vivo.

Embodiment 4：

Embodiment 4 describes the effect of skin micropin application (+)-saxitoxin.When by micropin conveying come testing stone During the analgesic efficacy of STX, all rats all use Isoflurane (2.5%) deep anaesthesia.Every male Sprague-Dawley The left and right cheek of rat (Harlan, 250g) is lost hair or feathers completely, it is thoroughly cleaned with ethanol afterwards.

In the case of deep anaesthesia, microneedle patch is applied to every rat.Tester is with the tweezers of a pair of blunt edges by one The skin of side cheek is pulled back, so that the skin-tightening of rat.Now, tester will promote the microneedle patch containing saxitoxin It is downwardly into skin.Equally, identical operating process is carried out on the cheek of opposite side；However, the paster plays the work of control With, and the not drug containing in micropin syringe needle.Each test group is made up of 6 rats that application of saxitoxin.

Microneedle patch is stopped about 20 minutes on rat skin, to ensure enough drug deliveries.During this period, greatly Mouse and is allowed it to recover in respective cage by the declines of Isoflurane.Once paster is removed, with regard to intraperitoneal injection 25% Urethane (1mg/kg), make rat light anaesthesia, so as to allow to carry out behavior measure in the case of no destructive motion.Open After beginning 20min, with 10min time interval and in ensuing 2.5h, every 30 minutes once within 30min time The contracting cheek reaction time (cheek withdrawal latencies) that repeat assessment is heated for lasting nocuity.Using adding Thermolamp (is set as 40V and is positioned at more than the cheek of rat 7 centimeters), and application radiant heat is concentrated to the left cheek of rat；Record Rat responds the time that thermostimulation is consumed.Same process is repeated in right cheek.Rat is not responded after heated 20 seconds, is considered Without response.

As a result：

The reaction that the skin treated with saxitoxin paster is heated to nocuity significantly (p ＜ 0.05, ANOVA) is less than Compare skin (Figure 13).Therefore, the thermal response reaction time observed is than with the treated high 2- of skin of control microneedle patch 2.5s, this represents that pain sensitivity is moderately reduced.

Embodiment 5：

5.1 can synthesize and modify saxitoxin and gonyatoxin.We have been realized in saxitoxin in laboratory With the synthesis of GTX-2/3, and the form of these unique toxins and the method (Figure 14) of function are modified.We are to STX What problem was analyzed, which be discussed in detail, has occurred in document.(referring to document：(a) Fleming, J.J., M.D.McReynolds, and J.Du Bois, (+)-saxitoxin：a first and second generation Stereoselective synthesis.J Am Chem Soc, 2007.129 (32)：p.9964-75.(b) Fleming, J.J.and J.Du Bois, A synthesis of (+)-saxitoxin.J Am Chem Soc, 2006.128 (12)： p.3926-7.) our research is it has been proved that the modification to primary carbamate unit (C13) in STX significantly changes this Compound is to TTX-sNa_VThe affinity of passage.Based on these data, we have been prepared for STX (NH derived from amine₂- STX), It can connect different prothetic group (such as fluorogen,¹⁸F- labels, biotin).The compound and the synthetic route of dependency structure are By height optimization, in order to prepare key intermediate with many grams of scale.We prepare Oregon- using these chemicals Green-STX and Cy5-STX conjugates, and these compounds have been tested to the rNa that is expressed in Chinese hamster ovary celI_V1.4 effect.Entirely Cell voltage pincers record shows that both molecules all retain to the Na_VIsoform has the affinity of middle nanomole level.Our mesh It is preceding to heterologous rNa_V1.7 have carried out electrophysiology measurement, and plan to use rNa_V1.8 as control, to rNa_V1.3 and TTX-r leads to Road is also combined research.

We have demonstrated that, NH₂- STX can be efficiently coupled into 4- fluobenzoic acids (SFB) n-hydroxysuccinimide On ester, so as to generate [19F] benzamide-STX.Measure and determine through full cell electrophysiology, our first generation PET medicaments Non-radioactive analogs are shown to rNa_V1.4 IC₅₀For 46 ± 7nM.Importantly, SFB is connected into NH₂- STX bar Part is suitable for generation should [18F] analog (see below).

Gonyatoxin (GTXs) is included with STX in structurally and functionally all closely related guanidine sulfate toxin family.This A little molecules are shown for TTX-s Na_VS has the low effect to middle nanomole level.It is interesting that our laboratories is qualitative Measurement display, has more slowly disengaging speed binding kineticses (off-rate binding compared to STX, GTX 3 kinetics).Because we can obtain these toxin by de novo formation, so having taken up similar step to prepare ammonia Carbamate derivative, and assess the influence that these new constructions are combined to passage.In addition, we have synthesized NH₂-GTX 3 Analog, and it is proved the reactive NHS esters generation single step chemical reaction (figure that this material can successfully with fluorescent dye 15).We are now residing unique status, except other those binding affinity and combination to passage of these toxin Dynamics change (i.e. slower disengaging rate kinetics, referring to following) non-natural derivatives beyond, also carry out STX and Internal comparative study (i.e. biodistribution, discharge rate, metabolism) between GTX 3 mark pattern.These poison can only be passed through The synthetic schemes from the beginning assembled of element can just make it possible that these are studied.

The STXs of 5.2 carbamates modification retains to Na_VNanomole affinity and can have longer effect hold The continuous time.

One key issue to be solved be whether available imaging agent by saxitoxin functionalization without interfering significantly with With reference to effect.We have been prepared for new STX fluorescent conjugate and PET conjugates by the such flow of design, should Flow make use of the property that STX derivatives such as 4 (Figure 15) can be used for selective chemical to be connected to electrophilicity ester reagent.For example, changing Compound 4, when being handled under gentle alkalescence condition with Cy5-N- hydroxysuccinimidyls acylimino (NHS) ester, only obtains coupled product. What the Connection Step was typically completed in less than 3h, and it is compatible with various NHS ester derivants, so that Oregon The synthesis of Green, Cy5 and p-19F- heterocyclic carbamate derivatives is possibly realized.Electrophysiology record determines these compounds all Retain to Na_V1.4 have the low effect to middle nanomole level.

We have observed that when Cy5-STX or natural STX is subcutaneously injected to the hind paw of murine, in anesthesia In phase (sensitiveness declines or pain threshold is improved), obvious difference is shown for mechanical irritation, between STX and Cy5-STX (Figure 16).For Cy5-STX, its local anaesthesia effect lasts 60h or than more than 20 times of STX this height.In order to explain medicine This significant difference on acting duration, it is believed that big lipophilicity Cy5 dye molecules may with plasma membrane intimate association, from And the effect of film " anchor " is effectively acted as, and STX structure divisions are then located in the collar extension of access opening.

5.3 [18F] benzamide-STX synthesis.

We are it has been shown that NH₂- STX can selectively be coupled to reactive NHS esters (such as [19F] -4- fluobenzoic acids) On.Recently, the coupling reaction is successfully replicated with [18F]-SFB, generates radiolabeled [18F] benzamide-STX (Figure 17).[18F]-SFB preparation details are as previously described.(referring to document：(a) Li, Z.B. etc., 18F-Labeled BBN-RGD Heterodimer for Prostate Cancer Imaging.J Nucl Med, 2008.49 (3)：p.453-61.(b) Wu, Z. etc., 18F-labeled mini-PEG spacered RGD dimer (18F-FPRGD2)：synthesis and microPET imaging of alphavbeta3 integrin expression.Eur J Nucl Med Mol Imaging, 2007.34 (11)：p.1823-31.) in pH 9.5 aqueous CH₃Carried out and NH in CN₂- STX coupling, and pass through Preparative HPLC carrys out purified product.All chemical reactions can by using modification GE TRACERlab FX-FN synthesis modules (GE Medical Systems；Positioned at Milwaukee, WI, USA city) semi-automatic completion.With sterilizing filter pair [18F] benzamide-STX purification fractions progress is degerming, so as to obtain final product.By for preassembled batch of material phial The asepsis injector of (batch vial) part is by [18F] benzamide-STX of preparation sample from prepared batch of material Taken out in phial.The aliquot is distributed, for carrying out following QC tests：Appearance test, radio chemistry and chemical purity, ratio Radioactive activity, aseptic, pyrogenicity, pH value, residual solvent analysis, radioactivity nuclear species identification (radionuclidic ) and bacterial endotoxin (lal test) identity.

5.4 [18F] benzamide-STX preliminary biodistribution research shows the stability of tracing in vivo agent.

After [18F] STX is injected intravenously into Mice Body into (n=3), obtain organ to determine radiolabeled life Thing credit cloth.Not it was observed that the related bone intake (0.05 ± 0.03%ID/gm) of statistics, this shows fluorobenzoyl amine groups De- fluorination will not occur in vivo.In addition, being found that 10.7 ± 8.9%ID/g in kidney, it appears that homaluria is main Tracer remove approach.As expected, in view of the dication of STX probes, does not detect spike in brain Agent (0.03 ± 0.02%ID/gm)；Thus, it is expected that the radioactive tracer Central nervous system does not influence.

5.5 in neuropathic pain rat model, [18F] benzamide-STX small animal position emission tomography (PET)s-MRI (micro- PET-MRI) Show that tracer is improved to the combination of nerve lesion sites.

Attempted Na for the first time at us_VWhen expression is imaged, to by the Sprague- for retaining neurotrosis (SNI) It is the neuropathic pain model accepted extensively that Dawely rats, which carry out [18F] benzamide-STX micro- PET-MRI, wherein SNI, It is related in 3 Main Branches of sciatic nerve 2 connects.(referring to document：For example, Decosterd, I.and C.J.Woolf, Spared nerve injury：an animal model of persistent peripheral Neuropathic pain.Pain, 2000.87 (2)：p.149-58.) strong soft tissue contrast is provided and outer due to MRI The profile of all nerve and DRGs, we make use of 7.0T toys MRI (to be available from our small animal imaging mechanism (Small Animal Imaging Facility)) produce sciatic nerve internal anatomy.In the specific example, animal by (inflammation has been crossed for quite a long time after having dissipated after surgery) is imaged when 5 weeks after damage.In isoflurane anesthesia Under, animal is firmly located on transferable fixator, mouse or rat are carried on fixed position by it, for the whole body Study following two imaging patterns：Micro- PET (the eXplore Vista microPET of GEHC (Suisna) company) and aobvious Micro- MRI (General Electric Co. Limited " 7T of MicroSigna 7.0 ").It is injected intravenously 500 μ Ci [18F] benzamide-STX Afterwards, 10min (20min after injection radioactive tracer) is imaged to rat with micro- PET, is then transferred to micro- MRI.It is fixed on Fiducial marker and animal on fixator make it possible to record micro- PET and micro- MRI data group jointly.Rat holder exists Without interference with imaging process in any discussed imaging pattern.

In figure 18, the micro- MR images of transverse direction (image on the left side) of the left sciatic injured level of femur are passed through The neuroma of expansion is shown, and the size of right sciatic nerves and diameter are normal.Middle image is in same observation The image for the micro- PET experiments of [18F] benzamide-STX that place is obtained, right figure shows two images recorded jointly.Enhancing Micro- PET signal appear in SNI generation neuroma vicinity.By contrast, failed to observe a signal in right neuron.Separately Outside, the increase (red arrow) of reflectivity tracer uptake amount is also shown in the soft tissue near injured nerve.In god Activity through being found beyond knurl may describes other changes related to neurotrosis, and this is probably the important set of pain experience Into part.[18F] the STX activity found beyond neuroma shows, the Na of the intact tissue near neurotrosis_VUp-regulation It is probably a key factor of chronic pain syndrome development.Other people are in injured nerve and its neighbouring undamaged nerve In observe it is such before pain events (pro-nociceptive event).For example, in impaired shin bone and normally Na is observed in undamaged nervus suralis in the cell body and periphery aixs cylinder and SNI models of fibular nerve_VBeta 2 subunit unit Up-regulation.(referring to document：For example, Pertin, M. etc., Upregulation of the voltage-gated sodium channel beta2 subunit in neuropathic pain models：characterization of Expression in injured and non-injured primary sensory neurons.J Neurosci, 2005.25(47)：p.10970-80.) determination is tested through von Frey in addition, what is observed in neurotrosis is increased [18F] benzamide-STX is associated with the tenderness sense enhancing that animal left foot is slapped.The center of image and the micro- PET signal in front It is related to the radioactive tracer substance excreted.

Embodiment 6：

We there is provided herein the syntheti c route of modification, and it results in the Na-ion channel blocker of C11- sulphations Derivative (is referred to as gonyatoxin).Our operation since alcohol 1 (Figure 19), it can come according to as described in embodiment 2 Prepare.The material is handled with electrophilic imidazole salts 2, carbonic acid phenyl ester is obtained.Primary amine and secondary amine can be added in the material, so as to produce The intermediate such as 4a and 4b of ammonifying carbamate modification.STX/GTX N21 substitutions analog is completed such as by the sequence of five steps 5a and 5b synthesis.

Experimental program and characterize data：

Alkene JVM.16 (792mg, 0.97mmol) 20mL THF solutions are cooled to -78 DEG C, and add tetrabutyl fluorine Change ammonium (1.16mL 1.0M THF solutions, 1.16mmol, 1.2 equivalents).The mixture is heated to 0 DEG C and stirred at such a temperature Mix 20min.Then, 20.0mL NH is added₄The reaction is quenched in Cl saturated aqueous solutions.The content is diluted with 40mL EtOAc Thing, and be transferred in separatory funnel.Collected organic layer and with 3x40mL EtOAc aqueous phase extracteds.By the organic extract of merging Dried and be concentrated under reduced pressure with magnesium sulfate.Purified with silica gel chromatography (2: 1 → 1: 2 hexanes/EtOAc), obtain JVM.21, be white Color solid (424mg, 75%).TLC R_f=0.33 (1: 2 hexane/EtOAc)；¹H NMR(CD₃OD, 400MHz) δ 6.28 (ddd, 1H, J=6.2,2.0,2.0Hz), 6.08 (ddd, 1H, J=6.2,2.2,2.2Hz), 4.67 (d, 1H, J=11.2Hz), 4.64 (d, 1H, J=11.2Hz), 4.43 (m, 2H), 4.35 (d, 1H, J=2.8Hz), 3.69-3.63 (m, 2H), 3.57 (dd, 1H, J =12.4,8.4Hz) ppm；¹³C NMR(CDCl₃, 125MHz) and δ 172.3,159.4,157.9,130.9,127.4,96.5, 94.1,83.1,78.4,61.7,55.5,55.2,54.5ppm；IR (film) n 3289,1616,1562,1379,1175,905, 841,730cm^-1。

Into ethanol JVM.21 (424mg, 0.73mmol) 7.3mL THF solutions add JVM.22 (297mg, 0.84mmol, 1.2 equivalents).The progress reaction is stirred at room temperature up to 23h, 15mL NaHCO is then added₃Saturated aqueous solution Reaction is quenched with 20mL EtOAc.Content is transferred in separatory funnel and organic phase is collected.With 2x 10mL EtOAc Aqueous phase extracted.The organic extract magnesium sulfate of merging is dried and is concentrated under reduced pressure.With silica gel chromatography (hexane → 1: 2 oneself Alkane/EtOAc) purifying, JVM.23 is obtained, is white solid (357mg, 70%).TLC R_f=0.52 (1: 2 hexane/EtOAc)；¹H NMR(CD₂Cl₂, 400MHz) and δ 9.57 (d, 1H, J=2.8Hz), 7.44-7.39 (m, 2H), 7.32-7.27 (m, 1H), 7.20-7.16 (m, 2H), 6.36 (ddd, 1H, J=6.2,2.0,2.0Hz), 6.04 (ddd, 1H, J=6.2,2.2,2.2Hz), 4.64 (s, 2H), 4.60-4.54 (m, 1H), 4.51-4.45 (m, 1H), 4.40 (dd, 1H, J=12,6.4Hz), 4.35 (d, 1H, J=3.2Hz), 4.31 (dd, 1H, J=12.0,5.2Hz), 3.98-3.93 (m, 1H) ppm.

To JVM.23 (53mg, 0.075mmol)) it is dissolved in 750 μ L CH₂Cl₂Solution in add tetradecylamine (150 μ L's 1.0M CH₂Cl₂Solution, 0.15mmol, 2.0 equivalents).The progress reaction is stirred at room temperature up to 5h, additional ten are then added Four alkanamines (300 μ L 1.0M CH₂Cl₂Solution).It is stirred for carrying out reaction at room temperature up to 12h, is then directly transferred to silica gel Post.With silica gel chromatography (hexane → 1: 2 hexanes/EtOAc) purify residue, obtain JVM.24, be white solid (22.1mg, 36%).TLC R_f=0.53 (1: 2 hexane/EtOAc)；¹H NMR(CD₂Cl₂, 400MHz) and δ 9.61 (br s, 1H), 6.33 (ddd, 1H, J=6.2,2.0,2.0Hz), 6.05 (ddd, 1H, J=6.2,2.2,2.2Hz), 4.98 (dd, 1H, J=5.6, 5.6Hz), 4.67-4.60 (m, 3H), 4.48-4.41 (m, 1H), 4.23 (dd, 1H, J=12.0,7.2Hz), 4.16 (dd, 1H, J =12.0,5.2Hz), 4.11 (d, 1H, J=4.0Hz), 3.87-3.81 (m, 1H), 3.17-3.10 (m, 2H), 1.53-1.44 (m, 2H), 1.27 (br s, 22H), 0.89 (t, 3H, J=6.8Hz) ppm.

¹H NMR(CD₃OD, 500MHz) δ 4.81 (s, 1H), 4.41 (dd, 1H, J=7.0,7.0Hz), 4.31 (dd, 1H, J =11.5,9.5Hz), 4.06 (dd, 1H, J=11.5,4.5Hz), 3.79-3.72 (m, 1H), 3.22 (dd, 1H, J=10.5, 7.0Hz), 3.18-3.06 (m, 2H), 1.51 (br s, 2H), 1.31 (br s, 22H), 0.92 (t, 3H, J=7.0Hz) ppm.

¹H NMR(D₂O, 500MHz) δ 4.83 (dd, 1H, J=7.5,7.5Hz), 4.63 (d, 1H, J=0.5Hz), 4.19 (dd, 1H, J=11.5,9.0Hz), 4.04 (dd, 1H, J=10.5,8.0Hz), 3.96 (dd, 1H, J=11.5,5.5Hz), (s, the 3H) ppm of 3.72 (dd, 1H, J=5.0,9.0Hz), 3.44 (dd, 1H, J=11.0,7.5Hz), 2.79 (s, 3H), 2.74.

Embodiment 7：

The bar of selective functionalization occurs for the R7 positions that we have been studied in the biguanides base framework for structure A descriptions of sening as an envoy to Part.The most common to be, these materials are prepared as the alkene JVM.16 or JVM.17 described in embodiment 2, but also can be from Dependency structure is prepared.The selective functionalization of R7 positions can be realized using various electrophilic reagents, and in described preparation After 11 β-OH-STX (embodiment 2) general way, product can be converted into by series of steps by the STX being deprotected completely Analog.Figure 20 is shown assembles acrylate and hydroxyl-methylene based moiety in this position, and the product is processed into STX R7- substitution analog.Figure 21 shows that the STX analogs and its electricity of a series of N7- substitutions prepared according to correlation technique are raw Pharmacological data, described electrophysiology data is related to the sodium channel isoform Na that the compound suppresses mammal_V1.4 energy Power.

Experimental program and characterize data：

To JVM.16 (100mg, 0.12mmol) 3.5mL CH₂Cl₂Added in solution 1,1,3,3- TMG (15 μ L, 0.12mmol, 1.0 equivalents).Mixture 1min is stirred, AW.1 (65mg, 0.21mmol, 1.7 equivalent) is then added.1.5h after The reactant is filtered with short silica gel plug.Purified with silica gel chromatography (2: 1 hexanes/EtOAc), obtain AW.2, be white solid (98mg, 71%).TLC R_f=0.31 (2: 1 hexanes/EtOAc)；¹H NMR(CDCl₃, 500MHz) and δ 9.52 (s, 1H), 7.65- 7.60 (m, 4H), 7.50-7.27 (m, 16H), 6.91 (s, 1H), 6.12 (d, 1H, J=6.0Hz), 5.82 (d, 1H, J= 6.0Hz), 5.30 (s, 2H), 4.91 (q, 2H, J=10.0Hz), 4.63 (s, 2H), 4.45-4.39 (m, 2H), 4.21-4.15 (m, 1H), 3.92-3.85 (m, 1H), 3.80-3.74 (br s, 1H), 3.67-3.58 (m, 2H), 3.43-3.35 (m, 1H), (s, the 9H) ppm of 3.33-3.26 (m, 1H), 3.06-2.99 (m, 1H), 1.06；¹³C NMR(CDCl₃, 125MHz) and δ 172.7, 169.9,160.1,155.7,152.5,135.8,134.5,133.8,132.1,130.8,130.5,130.1,129.3, 128.8,128.3,94.2,81.5,78.6,69.8,63.7,62.5,53.7,53.3,47.0,37.7,35.9,31.9,27.0, 22.9,19.3,14.4ppm.

By JVM.16 (30mg, 0.037mmol) 1.48mL CH₂Cl₂Solution is cooled to -20 DEG C, and then order adds 2- (13.5mg, 0.044mmol, 1.2 work as by tert-butyl group -1,1,3,3- TMGs (7.1 μ L, 0.037mmol, 1.0 equivalent) and AW.3 Amount).The reactant mixture is slowly heated to room temperature and 1.5h is stirred.Then content is directly crossed into silicagel column.Use silica gel color Spectrometry (2.5: 1 hexanes/EtOAc) is purified, and obtains AW.4 (30mg, 0.029mmol, 78%).TLC R_f=0.36 (2: 1 oneself Alkane/EtOAc)；¹H NMR(CDCl₃, 500MHz) and δ 9.46 (s, 1H), 7.90 (d, 2H, J=9.0Hz), 7.55-7.32 (m, 15H), 7.19 (s, 1H), 7.08 (d, 2H, J=9.0Hz), 6.21-6.14 (m, 2H), 5.24-5.14 (m, 3H), 4.60-4.49 (m, 3H), 4.43 (br s, 1H), 4.38-4.23 (m, 2H), 3.74 (br s, 1H), 3.60 (dd, 1H, J=10.5,6.5Hz), 3.47 (dd, 1H, J=10.5,7.5Hz), 0.93 (s, 9H) ppm.

¹H NMR(D₂O, 500MHz) δ 4.72 (s, 1H), 4.39 (dd, 1H, J=7.0,7.0Hz), 3.97 (dd, 1H, J= 10.0,8.0Hz), 3.86-3.78 (m, 1H), 3.77-3.63 (m, 4H), 3.70 (s, 3H), 2.40 (dd, 1H, J=10.5, 7.0Hz), 2.55-2.49 (m, 2H) ppm.

Embodiment 8：

Selective functionalization occurs for the R3 positions that we investigated in the biguanides base framework for causing structure A descriptions Condition.Figure 22 is shown assembles n-propyl in the position, and the C10- that the product is processed into GTX3 replaces analog.Also study The conditions of similarity of methyl group is assembled in C10 positions, and is directly illustrated in the experimental part below of the present embodiment.These processes STX and GTX R3- substitution analogs can be prepared by demonstrating us.

Experimental program and characterize data：

By JVM.15 (20mg, 0.023mmol) 460 μ L CH₂Cl₂Solution is cooled to -78 DEG C, and then order is added Me₂Zn (57 μ L 2.0M toluene solutions, 0.11mmol, 5.0 equivalents) and BF₃·OEt₂(6.61 μ L, 0.053mmol, 2.3 work as Amount).Reaction content is to slowly warm up to room temperature and stir 1h.Then, 1mL NaHCO is added₃Saturated aqueous solution makes the reaction It is quenched.The two phase liquid for being stirred vigorously acquisition reaches 15min.Then the mixture is diluted with 3mL EtOAc, and be transferred to point In liquid funnel.Collected organic layer and with 2x 3mL EtOAc aqueous phase extracteds.The organic extract of merging is dried simultaneously with magnesium sulfate It is concentrated under reduced pressure.With silica gel chromatography (gradient elution：Hexane → 2: 1 hexane/EtOAc) material that is isolated and purified with, obtain alkene JW.1, is white solid (11.7mg, 61%).¹H NMR(CD₃OD, 500MHz) δ 7.64-7.59 (m, 4H), 7.47-7.36 (m, 6H), 6.13 (dd, 1H, J=7.5,2.5Hz), 5.89 (dd, 1H, J=7.5,2.0Hz), 4.64 (d, 1H, J=13.5Hz), 4.60 (d, J=13.5Hz), 4.58-4.51 (m, 1H), 4.20 (d, 1H, J=3.5Hz), 3.79-3.75 (m, 1H), 3.72 (dd, 1H, J=13.0,5.5Hz), 3.52 (dd, 1H, J=13.0,10.0Hz), 1.55 (d, 3H, J=8.5Hz), 1.01 (s, 9H)ppm。

By JVM.15 (150mg, 0.171mmol) 3.42mL CH₂Cl₂Solution is cooled to -78 DEG C, and then order adds alkene Oxypropyl trimethyl silane (136 μ L, 0.86mmol, 5.0 equivalent) and BF₃·OEt₂(50 μ L, 0.39mmol, 2.3 equivalent).Make anti- Answer content to be to slowly warm up to room temperature and stir 1.5h.Then, 5mL NaHCO is added₃The reaction is quenched in saturated aqueous solution. The two phase liquid for being stirred vigorously acquisition reaches 15min.The mixture is diluted with 15mL EtOAc, and is transferred in separatory funnel. Collected organic layer and with 2x 10mL EtOAc aqueous phase extracteds.The organic extract magnesium sulfate of merging is dried and depressurized dense Contracting.The material being isolated and purified with silica gel chromatography (gradient elution: hexane → 2: 1 hexane/EtOAc), obtains alkene JW.2, is White solid (102mg, 70%).¹H NMR(CD₃OD, 500MHz) δ 7.64-7.58 (m, 4H), 7.46-7.36 (m, 6H), 6.15 (dd, 1H, J=7.5,2.5Hz), 5.95 (dd, 1H, J=7.5,2.5Hz), 5.84-5.72 (m, 1H), 5.23-5.14 (m, 2H), 4.61 (d, 2H, J=1.0Hz), 4.53-4.48 (m, 1H), 4.19 (d, 1H, J=3.5Hz), 3.78-3.70 (m, 2H), 3.54-3.48 (m, 1H), 2.99-2.92 (m, 1H), 2.68 (ddd, 1H, J=17.0,10.5,10.5Hz), 1.02 (s, 9H) ppm；¹³C NMR(CD₃OD, 125MHz) δ 171.5,160.2,157.9,135.7,135.6,134.1,133.5,132.4, 132.3,130.1,127.9,127.5,119.0,94.3,82.0,78.2,67.1,63.8,58.0,55.8,47.4,26.3, 18.8ppm。

In N₂In environment, to solid JW.2 (102mg, 0.12mmol) and Ru (PPh₃)₃Cl (1.1mg, 0.0002mmol, 0.01 equivalent) middle 1: the 1 toluene/EtOH for adding 12mL deoxidations.Hydrogen atmosphere is introduced, and the reactant mixture is stirred at room temperature 1h.Then, reactant is concentrated under reduced pressure.It is isolated and purified with silica gel chromatography (gradient elution: hexane → 2: 1 hexane/EtOAc) Material, obtain JW.3, be white solid (96mg, 94%).¹H NMR(CD₃OD, 500MHz) δ 7.67-7.62 (m, 4H), 7.49-7.40 (m, 6H), 6.28 (dd, 1H, J=6.0,2.0Hz), 5.95 (dd, 1H, J=6.0,2.0Hz), 4.67 (d, 1H, 11.5Hz), 4.64 (d, 1H, 11.5Hz), 4.52-4.48 (m, 1H), 4.21 (d, 1H, J=3.0Hz), 3.82-3.76 (m, 2H), 3.55 (dd, 1H, J=9.5,7.0Hz), 2.45-2.36 (m, 1H), 1.71-1.62 (m, 1H), 1.53-1.36 (m, 2H), 1.05 (s, 9H), 1.01 (t, 3H, J=7.5Hz) ppm；¹³C NMR(CD₃OD, 125MHz) δ 171.4,160.0,158.2, 135.7,135.6,134.8,132.4,130.0,127.93,127.90,127.0,81.6,78.3,67.2,63.9,58.2, 55.6,37.0,26.3,19.1,18.8,13.3,0.3ppm.

¹H NMR(D₂O, 500MHz) δ 4.35 (d, 1H, J=6.0Hz), 4.16-4.13 (m, 2H), 4.04 (d, 1H, J= 6.0Hz), 3.67-3.62 (m, 1H), 3.53-3.48 (m, 1H) 1.80 (br s, 1H), 1.63 (br s, 1H), 1.35 (br s, 2H), 0.86 (t, 3H, J=7.5Hz) ppm.

It should be understood that the ratio, concentration, quantity and other numeric datas in the application can be expressed as range format.Can The use of range format is that for convenience and simplicity, therefore, should understand in a flexible way to understand, it not only includes being used as model The numerical value for enclosing the limit and clearly describing, but also including all each numerical value comprising within the range or sub-ranges, as Clearly describe each numerical value or sub-range is the same.For example, concentration range " about 0.1% to about 5% ", it should be interpreted that not Instrument includes about 0.1% to about 5% concentration that clearly describes, but also including in the scope each concentration (such as 1%, 2%th, 3%, 4%) and sub-range (such as 0.5%, 1.1%, 2.2%, 3.3%, 4.4%).In embodiments, term " about " can Conventional rounded up including what is made according to the effective digital of the numerical value.In addition, phrase " about ' x ' to ' y ' " is included " about ' x ' To about ' y ' ".

Variations and modifications can be carried out to the embodiment above.It is public that all such modifications and variations are included in the application Open in the range of being protected with appended claims.

Claims (21)

1. it is any one in a kind of compound, its isomer, its dynamic isomer or these materials with following structure A The pharmaceutically useful salt of any one in the pro-drug of person or these materials：

Wherein R1 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,= O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、- SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AGroup such as what follows can independently be：Hydrogen, It is halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, complete It is fluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, miscellaneous Cyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), Nitro, oxa-, oxo, sulfonyl, sulfonamido and sulfone；

Wherein R2 is selected from the group being made up of following groups：Hydrogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo- Substituted alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) Alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, heteroaryl (alkyl), oxa-, oxo, sulfonyl, Sulfonamido, sulfone ,-OR_A,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、- SO₂R_A、-NO₂With-C (R_A)₃, wherein each R_AHydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, halo can independently be Alkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, It is (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido and sulfone；

Wherein R3 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、-SO₂R_A、- N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AHydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyanogen can independently be Base, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, take For phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, Shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfenyl ammonia Base and sulfone；

Wherein R4 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、-SO₂R_A、- N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AHydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyanogen can independently be Base, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, take For phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, Shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfenyl ammonia Base and sulfone；

Wherein R5 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfate radical, sulfonamido, Sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、- SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AHydrogen, halogen, alkane can independently be Base, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, Cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, miscellaneous cyclic group Group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxygen Miscellaneous, oxo, sulfonyl, sulfonamido and sulfone；

Wherein R6 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfate radical, sulfonamido, Sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、- SR_A、-SOR_A、-SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AHydrogen, halogen, alkane can independently be Base, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, Cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, miscellaneous cyclic group Group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxygen Miscellaneous, oxo, sulfonyl, sulfonamido and sulfone；And

Wherein R7 is selected from the group being made up of following groups：Hydrogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, dihalo- Substituted alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, (substituted-phenyl) alkyl, virtue Base, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, Asia Sulfonamido, sulfone ,-OR_A,=O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、- CN、-SO₂R_A、-NO₂With-C (R_A)₃, wherein each R_ACan independently be hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, Haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted benzene Base, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, protected Shield (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido and Sulfone,

Precondition is：Compound with structure A is not to be selected from saxitoxin, N-STX, gonyatoxin, spot Compound in compound shown in sufficient toadpoison AB and Fig. 1.

2. it is any one in a kind of compound, its isomer, its dynamic isomer or these materials with following structure B The pharmaceutically useful salt of any one in the pro-drug of person or these materials：

Wherein R1 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,= O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、- SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AHydrogen, halogen, alkyl, alkenyl, alkynes can independently be Base, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (ring Alkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulphur Acyl group, sulfonamido and sulfone,

Precondition is：Compound with structure B is not to be selected from saxitoxin, N-STX, gonyatoxin, spot Compound in compound shown in sufficient toadpoison AB and Fig. 1.

3. it is any one in a kind of compound with following structure Cs, its isomer, its dynamic isomer or these materials The pharmaceutically useful salt of any one in the pro-drug of person or these materials：

Wherein R1 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,= O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、- SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AHydrogen, halogen, alkyl, alkenyl, alkynes can independently be Base, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (ring Alkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulphur Acyl group, sulfonamido and sulfone,

Precondition is：Compound with structure C is not to be selected from saxitoxin, N-STX, gonyatoxin, spot Compound in compound shown in sufficient toadpoison AB and Fig. 1.

4. it is any one in a kind of compound, its isomer, its dynamic isomer or these materials with following structure D The pharmaceutically useful salt of any one in the pro-drug of person or these materials：

Wherein R1 is selected from the group being made up of following groups：Hydrogen, halogen, alkyl, alkenyl, alkynyl, alkoxy, cyano group, haloalkyl, Dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (cycloalkyl) alkyl, substituted-phenyl, (substitution Phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (singly take Generation) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulfonyl, sulfonamido, sulfone ,-OR_A,= O ,-C (=O) R_A,-OC (=O) R_A,-OC (=O) OR_A,-OC (=O) N (R_A)₂、-CO₂R_A、-CN、-SCN、-SR_A、-SOR_A、- SO₂R_A、-NO₂、-N(R_A)₂、-NHC(O)R_AWith-C (R_A)₃, wherein each R_AHydrogen, halogen, alkyl, alkenyl, alkynes can independently be Base, alkoxy, cyano group, haloalkyl, dihalo alkyl, tri haloalkyl, whole haloalkyl, perfluoroalkyl, cycloalkyl, (ring Alkyl) alkyl, substituted-phenyl, (substituted-phenyl) alkyl, aryl, heteroaryl, heterocyclic type group, heterocyclic group, alkanoyl, (monosubstituted) amino, shielded (monosubstituted) amino, (disubstituted) amino, heteroaryl (alkyl), nitro, oxa-, oxo, sulphur Acyl group, sulfonamido or sulfone,

Precondition is：Compound with structure D is not to be selected from saxitoxin, N-STX, gonyatoxin, spot Compound in compound shown in sufficient toadpoison AB and Fig. 1.

5. a kind of compound, it is included connects with label, oligonucleotides, protein, lipid, steroids, antibody or antibody fragment Compound described in any one in the claim 1-4 connect.

6. compound as claimed in claim 5, wherein compound described in any one passes through altogether in described claim 1-4 Valence link is connected with label, oligonucleotides, protein, lipid, steroids, antibody or antibody fragment.

7. compound as claimed in claim 5, it includes oligonucleotides, protein, lipid, steroids, antibody or antibody piece Section, wherein the oligonucleotides, protein, lipid, steroids, antibody or antibody fragment structure division can be effectively by Saxidomus Toxin analog is directed to sodium channel, specific voltage-gated sodium channel isoform or expression Na_VCell type vicinity.

8. compound as claimed in claim 5, wherein the label is selected from the group by following material composition：The same position of radioactivity Element, fluorescence structure part, chemiluminescent structure part, enzyme, antibody, antibody fragment, magnetic-particle and quantum dot.

9. a kind of pharmaceutical composition, its include compound in claim 1-8 described in any one, its isomer, its The pharmaceutically useful salt of any one in the pro-drug or these materials of any one in dynamic isomer or these materials.

10. compound, its isomer, its dynamic isomer or these materials in claim 1-9 described in any one In any one pro-drug or these materials in the pharmaceutically useful salt of any one be used for need receive treatment it is tested The purposes that person is treated, its consumption is that can effectively treat the amount of the subject.

11. compound, its isomer, its dynamic isomer or these materials in claim 1-9 described in any one In any one pro-drug or these materials in the pharmaceutically useful salt of any one be used for reduce subject neuron live Property or cause subject loosening all muscles purposes, its consumption be can effectively reduce the subject neuronal activity or Person can effectively make the amount of the loosening all muscles of the subject.

12. purposes as claimed in claim 10, wherein the subject suffers from the enhanced disease of voltage-gated sodium channel.

13. purposes as claimed in claim 12, wherein the enhanced disease of the voltage-gated sodium channel is selected from by following disease In the group of composition：Acute Pain, anal fissure, arthritis, backache, chronic ache, dentistry pain, fibromyalgia, arthralgia, inclined head Bitterly, neuropathic pain, postoperative pain, herpes zoster, tonicity head after cervical pain, neuropathic pain, labor pain, herpes zoster Pain or trigeminal neuralgia pain, blepharospasm, cancer, arrhythmia cordis, epilepsy, focal dystonia, ephidrosis, muscle convulsion Contraction and atony of bladder.

14. purposes as claimed in claim 10, wherein the subject is by pain.

15. purposes as claimed in claim 14, wherein the pain is selected from：Acute Pain, anal fissure pain, arthritis ache, Backache, blepharospasm pain, cancer pain, chronic ache, dentistry pain, fibromyalgia, arthralgia, antimigraine, neck pain Bitterly, neuropathic pain after splanchnodynia, neuropathic pain, labor pain, herpes zoster, postoperative pain, sympathetic nerve maintenance pain, Herpes zoster pain, tension headache, trigeminal neuralgia pain, myositis pain, musculoskeletal pain, pain in the back, sprain and pull pain, To functional intestines problem such as non-ulcer dyspepsia, the NCCP pain related with irritable bowel syndrome, with The related pain of myocardial ischemia, toothache and pain caused by dysmenorrhoea.

16. compound, its isomer, its dynamic isomer or these materials in claim 1-9 described in any one In any one pro-drug or these materials in the pharmaceutically useful salt of any one be used for diagnose subject's illnesses Purposes, its consumption is that the enhanced disease of voltage-gated sodium channel effectively can be positioned at into specific region in subject's body Amount.

17. compound, its isomer, its dynamic isomer or these materials in claim 1-9 described in any one In any one pro-drug or these materials in the pharmaceutically useful salt of any one be used for subject is imaged and The subject is set to undergo the purposes of imaging process, its consumption is that can effectively detect the compound in imaging process The amount of position in subject's body.

18. compound, its isomer, its dynamic isomer or these materials in claim 1-9 described in any one In any one pro-drug or these materials in the pharmaceutically useful salt of any one be used to treat the purposes of wrinkle, it is used Measure that the amount of wrinkle can be effectively mitigated or eliminated.

19. a kind of method for preparing saxitoxin analog, including：

(i) guanidine of nine-atomic ring is made to react generation C13-Troc carbonate；

(ii) in the presence of a lewis acid, close the guanidine ring in the product of (i)；And

(iii) product of (ii) is aoxidized and is deprotected,

So as to generate saxitoxin analog.

20. a kind of method for preparing saxitoxin analog, including：

(i) 1- serine methylesters are made to react to form aldehyde；

(ii) aldehyde of (i) and amine are condensed；

(iii) product of (ii) is made to react and effectively close ring and generate carbamide compound；

(iv) product of (iii) is made to react in such a process, the process includes allyl deprotection and protected and isothiourea Formation；And

(v) by the product amination of (iv),

So as to generate saxitoxin analog.

21. the saxitoxin similar compound as described in any one in claim 1-9, wherein the effect of the compound Duration Ratio saxitoxin is long.