CN1187773A - Method for treatment of pain - Google Patents
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- CN1187773A CN1187773A CN96194784A CN96194784A CN1187773A CN 1187773 A CN1187773 A CN 1187773A CN 96194784 A CN96194784 A CN 96194784A CN 96194784 A CN96194784 A CN 96194784A CN 1187773 A CN1187773 A CN 1187773A
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Abstract
An improved method for the treatment of intractable pain is provided by the present invention. The improved method is the intrathecal administration of a therapeutic composition comprising elcatonin and a pharmaceutically acceptable carrier. Intrathecally administered elcatonin produces more potent and prolonged analgesia than elcatonin/calcitonins administered by other routes (e.g., intravenous, intramuscular injection) and/or opiates administered by different routes (including epidural or intrathecal administration).
Description
Invention field
Invention field is a treatment human pain, particularly intractable pain.Specifically, the invention provides elcatonin in the purposes in the pharmaceutical preparation of intrathecal route administration.
Background of invention
Known calcitonin is effective on the human pain of treatment, this pain results from diseases such as including, but is not limited to osteoporosis, scleromalacia, hallucination limbs pain, reflexive sympathetic nerve malnutrition, fracture, ankylosing spondylitis and malignant metastatic tumor of bone and (for example sees Gennari ﹠amp; Agnusdei (1988): Current Therapeutic Research 44:712-721).Calcitonin falls blood calcium because of it and serum phosphate lowering hydrochlorate effect is used in the therapeutic combination.Calcitonin also is used to increase sclerotin.
Anesthetics (as morphine) also is used to treat human chronic pain, and it has the shortcoming of addiction, can produce toleration to the non-toxic dose of morphine.
The patient that can not rely on conventional analgesia therapy is carried out analgesia therapy, demand is arranged technically for a long time always.Such pain therapy do not have significant side effects or toxicity; And also the pain therapy of toleration does not appear in requirement.The invention provides effective treatment, particularly to the patient's (as the patient of intractable pain that terminal cancer is accompanied) that suffers from chronic and intractable pain treatment to pain.
Summary of the invention
The invention provides the improved method of treatment mammal pain (particularly treating human intractable pain).In the present invention, provide elcatonin to the patient who suffers from pain through intrathecal drug delivery.Elcatonin is dissolved in the excipient (as sterile saline) that is suitable for intrathecal injection.That preferable is the human serum albumin that medicine further comprises the about 0.03-0.1%mg/ml of concentration.Dosage range is the about 15IU elcatonin of about 0.2-/kg body weight, and preferable is the about 6.4IU elcatonin of about 1.6-/kg body weight.Dosage be advisable to be mixed with the intrathecal injection normal volume (being preferably the about 2.0ml of the about 0.5-of single agent intrathecal injection).Elcatonin by the described dosage of this description with single agent or multi-agent through the intrathecal route administration.The multi-agent administration can reach 24-48 hour at interval.Can adopt intrathecal route by injecting or with continuously or the administration of pulsed administration pump.
The elcatonin intrathecal drug delivery produces powerful, persistent analgesic effect, without significantly nonconforming side reaction, as addiction, does not have tangible toleration to take place.Pain therapy of the present invention helps to suffer from the patient of intractable and serious pain (as terminal cancer) especially.
The simple description of accompanying drawing
Fig. 1 illustration gives elcatonin through intrathecal route and prolongs hot plate incubation period in the dose dependent mode.1 hour mensuration hot plate incubation period (second) after the elcatonin administration.
Fig. 2 illustration in sheath, give the elcatonin prolong rats hot plate preclinical effect of dosage from the 0.2-3.6IU/kg body weight (+-0.2IU/kg ,-★-0.8IU/kg ,-■-1.6IU/kg ,-*-2.4IU/kg ,-◆ and-3.6IU/kg ,--normal saline contrast).
Fig. 3 A and 3B provide and give elcatonin (a) dosage in the sheath is that 2.4-6.0IU/kg and morphine (b) dosage are the comparison of rat hot plate prolongation of latency behind the 37.5-125.0mg/kg.In these researchs, be 40 seconds maximum incubation period.
Fig. 4 provide through intrathecal route give the low dosage elcatonin (2.4IU/kg ,-●-), the hot plate prolongation of latency of morphine (37.5mg/kg ,--) or normal saline (▲-) and the comparison of action time.
Fig. 5 provides the hot plate prolongation of latency of dosage elcatonin in intrathecal route gives (4.4IU/kg ,--), morphine (75.0mg/lg ,-■-) or normal saline (▲-) and the comparison of action time.In these researchs, be 40 seconds maximum incubation period.
Fig. 6 provide through intrathecal route give high dose elcatonin (6.0IU/kg ,--), morphine (125mg/kg ,-●-) or the hot plate prolongation of latency of normal saline (■-) and the comparison of action time.
Give elcatonin that dosage is the 4.4IU/kg body weight (■-) or normal saline (zero-) or false injection (◆-) the preclinical prolongation of back hot plate in Fig. 7 the is illustrated in 5 days repeatedly sheath.Vertical arrow is represented the time of rat injection.Measured hot plate incubation period in 0,1,3 and 24 hour in injection back the 1st, 3 and 5 day of research.
Giving 10 single agent (once a day) dosage in Fig. 8 illustration sheath is 4.4IU/kg elcatonin (■-) or normal saline (zero-) or false injection
The preclinical prolongation of back hot plate, that does not handle is spaced apart 48 hours.Vertical arrow is represented the time of rat injection.Research the 1st, 3,5,7,9 and measured hot plate incubation period in ll days in 0,1,3 and 24 hour in injection back.
Fig. 9 is illustrated in the preclinical prolongation of elcatonin (dosage is 4.4IU/kg) back hot plate that repeatedly contained HSA (0.1%) in 5 days in the sheath.Measured hot plate incubation period in 0,1,3 and 24 hour in injection back the 1st, 3 and 5 day of research.
The detailed description of invention
U.S. Patent No. 4,086, the described Elcatonin of 22l on Mays 3rd, 1976 such as (application) shumpei Sakakibara has following structure:
Elcatonin is replaced by amino suberic acid bridge from different N-terminal cysteine residue and the 7th amino acid whose positions of being of clcatonin, and it provides the carbon-to-carbon bridge to replace the disulfide bond between the cysteine residues in the natural calcitonin molecule. Other molecule that contains amino suberic acid bridge is in U.S. Patent No. 4,086, description arranged in 221, is incorporated herein by reference. The amino acid sequence of eel, people, pig, salmon and rat calcitonin is respectively sequence 1,2,3,4 and 5. It is terminal and the 7th cysteine also obtains quite stable, effective peptide composition eases pain to replace N-with amino suberic acid bridge in the calcitonin of sequence 2-5. The U.S. Patent No. 4,977,139 (Yamada etc., application on October 25th, 1989) of drawing in this article for reference is described the Elcatonin waterborne compositions to light, heat and vibrational stabilization property improvement.
U.S. Patent No. 4,977,139 (Yamada etc., applications on October 25th, 1989), No.5,118,667 (Adams etc., applied on May 3rd, 1991) and 4,758,550 (Cardineaux etc., December in 1986 application on the 3rd) as if disclose calcitonin and the purposes of elcatonin compositions on treatment all kinds bone and/or painful diseases, but none open elcatonin intrathecal injection is used for the treatment of intractable pain.
As if Miseria et a1. (1989) Tumori (Italy) 75:183-184 discloses the salmon calcitonin epidural administration and has been used for the treatment of intractable pain, but it seems that data do not supported it to be used as analgesics.
As used herein, analgesic effect refers to that the result is insensitive to pain, or the reduction of the sensation of the pain sensation or painful or destructive stimulus, and does not lose consciousness.Analgesia and anti-injury sensation are used as synonym in this application.
As used herein, intractable pain is to produce toleration and constant pain to the nullvalent pain of conventional analgesics and/or to opioid analgesics (as morphine).Intractable pain can be include, but is not limited to cancer, fracture, osteoporosis, cut-out postoperative pain and some other disease with.
Rat is the preferable animal model through the test-compound of intrathecal route administration, and the rat hot plate test of measuring analgesic activity is the acceptable model system of potential effective pain therapy.
(see embodiment 8,9 as disclosed herein; Table 1,2), measure in the model system of analgesic at the rat hot plate, used whole elcatonin dosage all produce powerful, persistent anti-injury sensation, in injection demonstration effect in back 15 minutes, reach peak value in the time of 30-60 minute, last till back 6 hours of injection at least.Adopt hot plate test, the normal analgesic activity that produces observable maximum of used elcatonin dosage is therefore at hot plate no significant difference incubation period with scope gained behind the elcatonin of 0.8-3.6IU/kg.At used lowest dose level (0.2IU/kg), observe lower slightly reaction.Yet, use than the low dosage elcatonin even also can obtain medium and MIN anti-injury that dosage relies on mode and feel and react.
Table 1
Analgesic activities (hot plate incubation period) (mean value standard error) behind the single agent injection of the rat elcatonin
*P≤0.05 student ' s t-test
*P≤0.05 student ' s t-test
| Dosage IU/kg | Number of rats | Hot plate incubation period (second) | ||||||
| Time after the administration (hour) | ||||||||
| ????0 | ????0.25 | ????0.5 | ????1 | ????3 | ????6 | ????24 | ||
| ????0 | ??21 | ??11.2±0.5 | ??11.7±0.9 | ??11.5±0.6 | ??12.2±0.8 | ??11.8±1.1 | ??11.1±0.7 | ??11.5±0.5 |
| ????0.2 | ??11 | ??10.6±0.8 | ??13.0±1.1 | ??12.9±0.7 | ??12.7±0.9 | ??13.1±0.7 | ??12.1±0.5 | ??10.8±0.7 |
| ????0.8 | ??13 | ??9.8±0.8 | 15.1±1.5 ** | 15.0±1.5 ** | 15.2±1.3 ** | 20.0±2.8 ** | ??12.3±1.0 | ??11.7±1.0 |
| ????1.6 | ??13 | ??9.2±0.5 | ??16.4±1.2 ** | 16.0±1.0 ** | 16.4±1.1 ** | 15.1±0.9 ** | 13.6±0.8 ** | ??10.9±0.4 * |
| ????2.4 | ??13 | ??9.2±0.7 | 20.0±1.5 ** | 20.1±1.9 ** | 19.5±1.8 ** | 21.8±2.1 ** | 12.6±1.2 * | ??10.31±1.2 |
| ????3.6 | ??12 | 10.4±0.9 ** | 19.4±1.6 ** | 21.4±1.8 ** | 23.5±2.5 ** | 19.5±1.0 ** | 15.7±1.3 ** | ??10.2±1.0 |
Measure in the test data of analgesic activity at rat hot plate described herein, the intrathecal injection elcatonin is compared with the intravenous injection elcatonin, obtains unexpected anti-injury and feels improved result (seeing this paper embodiment 7).Shown in embodiment 9 and Fig. 6-8, in the rat hot plate test, the intrathecal injection elcatonin is compared with the intrathecal injection morphine, also produces surprising pain and improves.In addition, analgesic activities at morphine and elcatonin intrathecal drug delivery is relatively gone up, elcatonin demonstrates in the rat hot plate test has bigger analgesic activities, and the analgesic effect of elcatonin also demonstrates the persistent period (seeing this paper embodiment 9 and Fig. 3-6) greater than morphine.
The following examples are not in order to limit described herein and scope claimed invention for the purpose of setting forth provides.All researchs are all carried out according to the rules (21 C.F.R.58) of FDA GLP.The modification of program of conspicuous any routine of carrying out from invention disclosed herein practice and deriving all within the scope of the invention concerning those of skill in the art.
Embodiment
Embodiment 1: experimental animal
Rat is the U.S. and international drug administration agencies about one of preferred species that are used for the test compound analgesic activities.Historical information about the Sprague-Dawley rat can obtain from disclosed document.(Oregon WI), makes it that laundering period of 7 days be arranged before being used to test to bull Sprague-Dawley rat available from Sasco Inc..After receiving rat, after 1 day and 7 days or before being assigned to experimental group it is weighed.Rat is assigned in the experimental group randomly handles.Body weight obviously is different from this group average weight, display abnormality changes or shows that any animal of physical disease is not all used on body weight.The test rat is in the 320-420g scope.
Embodiment 2: the care of animal
The care of animal is according to criterion and the laboratory animal protection of resources of animal protection regulations and guide for use (the Guide for the Care and Use of Laboratory Animal Resoures of United States Department of Agriculture (USDA); NationalResearch Council; DHHS; Publication No.NIH 85-23,1986).Rat is placed on the indoor of environment control, keeps temperature in 18-26 ℃, 18-20 ℃ usually, relative humidity 40-70% per hour carries out room air exchange more than 10 times.Keep 12 hour daily cycle every day.To animal supply arbitrarily tap water and food (Agway Rat Chow, Agway).The inventor do not find may to test dysgenic food is arranged or supply water on any pollution.
Twice weekly (or more diligent, if desired) from cage, remove Excreta.The per two weeks sterilization of cage and tableware once can be more diligent as needs.
In case intubate, rat is promptly closed in Rotating Stainless Steel Cage to avoid intubate to be damaged by the companion separately.Every rat is distinguished by the numbering of the uniqueness on its cage.
Embodiment 3: the preparation of storing solution
Elcatonin ([Des-Cys
1, ASU
7]-calcitonin, eel; Carbacalcitonin) derive from Bachem, (Torrance, CA), lucifuge is stored in-20 ℃ to Inc..Be used for storing solution (1mg/ml, in the sterilization isotonic saline solution, American Pharmacopeia) fresh preparation every day of the elcatonin of analgesic activities test behind the intrathecal injection and remain in the ice.Carry out suitable dilution before facing injection.
The sterilization isotonic saline solution (0.9%w/v NaCl, American Pharmacopeia level) derive from Sigma Chemical Co. (St.Louis, MO).
The human serum albumin derive from Sigma Chemical Co. (St.Louis, MO).Storing solution is 25% (w/v).
Morphine sulfate derives from Mallinckrodt, and Inc. (St.Louis, MO).Intrathecal injection morphine storing solution preparation every day (10mg/ml is in the sterilization isotonic saline solution) also remains on ice.Carry out suitable dilution before facing injection.
Embodiment 4: the nerve operation/intubation that is used for intrathecal drug delivery
Test preceding 5～7 days with rat anesthesia (Nembutal, 30mg/kg, lumbar injection) back operation implantation sheath interpolation pipe.(polyethylene tube PE10) inserts through the occipito-axial ligaments crack each pipe, and intubate is passed subarachnoid space 7.5cm, it is not held be positioned at waist and expand a lateral margin.Each pipe is fixed on the cranium with cyanoacrylate adhesive, closes cranium with wound clamp.Free-end heat-sealing with each pipe; When preparing, introduce each compositions with the Hamilton syringe needle that inserts the sealing intubate through the intrathecal route injectable composition.Intubate implant have before the test of back 5-7 days the time chien shih its from operation process, recover fully.
Embodiment 5: the evaluation of analgesic activities
Hot plate test (Woolfe and MacDonald (1944) J.Pharmacol.Exp.Ther.80:300) with the measurement analgesic activity of standard is estimated pain sensitivity.Hot plate reaction system is placed on rat on 55 ℃ of copper hot plates that are encapsulated in the lucite cylinder and measures.To be put on the hot plate and lick metapedes or the interval (in second) that jumps out of between the hot plate is defined as " hot plate incubation period ".If animal does not take place hot plate is worked the reaction of licking metapedes or jumping out of hot plate, after 40 seconds, take out to avoid sufficient tissue injury, its hot plate is decided to be 40 seconds incubation period.
Because the specification requirement of these experiments, test is no more than 8-9 rat in given one day.The enough animal of each experimental condition research is to obtain 10-11 valuable rat.In test day, animal is carried out the reaction of baseline hot plate, accept to be tried injecting of analgesics or normal saline (contrast) then.Revalued the hot plate reaction behind test-compound or the excipient tester intrathecal injection in 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours and 24 hours.
Embodiment 6: dosage
The elcatonin of all dosage is all to inject form administration.In single agent administration research, tested dosage in the following elcatonin sheath: 0.2,0.8,1.6,2.4,3.6,4.4,5.2 and 6.0IU/kg or 0.03,0.13,0.25,0.39,0.58,0.71,0.84 and 0.97 μ g/kg.The elcatonin intravenous injection dosage of test is 10.0,30.0,100.0 and 300.0IU/kg or 1.61,4.83,16.1 and 48.3 μ g/kg.1 μ g elcatonin is equivalent to about 6.2IU.
Embodiment 7: the administration volume
All intrathecal injection dosage all give with the volume of 37.5 μ l/kg, use 10 μ l/kg normal saline flushing intubate (single agent research) then or use 8 μ l normal saline flushing intubate (multi-agent research) then with 10 μ l volume administrations.All intravenous injection dosage all give with the volume of 0.5ml/kg.
Embodiment 8: in the rat analgesia determination test to the dose response of intrathecal injection elcatonin
In this research, a certain dosage range that intrathecal route gives elcatonin has been tested in analgesia in the determination test, determining to cause analgesic minimum and peak value effective dose and definite dose response.Being subjected to amount of reagent is 0.2,0.8,1.6,2.4 and 3.6IU/kg.Inject required dosage and clean intubate with 37.5 μ l/kg volumes through intrathecal route with 10 μ l saline.Reaction behind establishment of base line (before the administration) and injectable drug or the excipient when 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours and 24 hours.Each dosage elcatonin is with 11-13 animal, with 21 control vehicle (saline) animal in the research.
Use body weight to be 113 of the bull rat of 350-400g sums.As described in embodiment 4, the sheath interpolation pipe is implanted in operation.Operation is being had 3 rats deaths to before trying compositions after implanting the sheath interpolation pipe.The 4th rats death arranged behind intrathecal injection 0.8IU/kg elcatonin in 24 hours.
Originally studies have shown that giving elcatonin through intrathecal route increases hot plate incubation period (see Table 1, Fig. 1) in the dose dependent mode.Anti-injury feel acts on injects onset in back 15 minutes, occurs preclinical statistics significant prolongation between 0.25 hour and 3 hours.Injecting at 3 higher dosage also had significantly anti-injury feel effect in back 6 hours.In the great majority tests, the hot plate of injecting back 24 hours animals is got back to baseline (see Table 1, Fig. 2) incubation period.
Data have been made significance analysis, analyzed the analgesic activities of each dose drug in seclected time with each point counting (average and standard deviation).Calculate the nonparametric statistics value, provide the significance level of difference between matched group and the test group.
The reaction that minimum test dose (0.2IU/kg elcatonin) does not produce significance,statistical, but 0.8 and 1.6IU/kg dosage cause significant anti-injury feel effect and 2.4 and 3.6IU/kg dosage cause powerful anti-injury to feel effect (Fig. 2).Matched group does not change baseline response only for saline.
Embodiment 9: the analgesic activities of intrathecal injection elcatonin and intrathecal injection morphine relatively
This research is the analgesic activities of morphine and elcatonin relatively, each medicine all on rat animal model through the intrathecal route administration.
In this research with 96 rats of body weight from 320g to 420g.As mentioned above, implant intubate and treat its recovery.Elcatonin dosage is 2.4,3.6,4.4,5.2 and 6.0IU/kg (being respectively 0.39,0.58,0.71,0.84 and 0.97 μ g/kg).Morphine sulfate is with 37.5,75.0 and 125.0 μ g/kg dosed administrations.Medicine or saline all adopt the Hamilton syringe needle that inserts intubate to give, and dose volume is 37.5 μ l/kg, then clean intubate with 10 μ l saline.Establishment of base line (before the administration) and give medicine or excipient after the reaction of 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours and 24 hours.
Elcatonin (Fig. 3 A) and morphine (Fig. 3 B) all prolong hot plate incubation period, and showing has analgesic effect (table 2).All are subjected to the elcatonin of amount of reagent all to produce powerful and long anti-injury feel effect, appearance effect in back 15 minutes of injection, and peaking in the time of 30-60 minute in the injection back, continues at least 6 hours (table 2, Fig. 3 A).Owing to use hot plate test, used elcatonin dosage often produces maximum detectable analgesic activity, so give hot plate no significant difference incubation period of the elcatonin gained of 3.6-6.0IU/kg scope dosage.Obtain smaller reaction when using lowest dose level elcatonin (2.4IU/kg or 0.39 μ g/kg).Yet the result of embodiment 8 shows, uses the medium and minimal anti-injury feel reaction that can obtain the dose dependent mode than the low dosage elcatonin.
Morphine prolongs hot plate incubation period in the dose dependent mode, and the analgesic appears in back 15 minutes of the injection (table 2, Fig. 3 A).Yet the analgesic persistent period seems and is shorter than elcatonin, injects and obviously shortens back 3 hour incubation period, and the injection back was got back to baseline hot plate incubation period in about 6 hours, and is especially like this than low dosage.And when this studied employed dosage, the analgesia peak level that animal reached of injection morphine generally was lower than the level (table 2, Fig. 3 A, 3B) of injection elcatonin animal.
In order to compare the analgesia intensity of elcatonin and morphine, convert dosage to nmol/kg.On several time points, this two medicine is made dose-effect curve with assessment dose,equivalent (table 2).Elcatonin 0.12nmol/kg (2.4IU/kg or 0.39 μ g/kg) injection produced the hot plate incubation period that is equivalent to morphine sulfate 98.84nmol/kg (75.0 μ g/kg) in back 30 minutes, and 0.18nmol/kg elcatonin (3.6IU/kg or 0.58 μ g/kg) is equivalent to morphine sulfate 164.73nmol/kg (125.0 μ g/kg) approximately.Similarly intensity sees back 1 hour (table 2) of injection.Yet, anti-injury in 6 hours feels that the specific activity elcatonin reduces behind the injection of morphia, therefore, at this moment between on the point, elcatonin is produced than the whole stronger hot plate analgesic activities (table 2) of amount of reagent (49.42-164.73nmol/kg) that are subjected to of morphine sulfate by amount of reagent (0.12-0.30nmol/kg) all.
Table 2
The analgesic activities of elcatonin and morphine behind the single agent intrathecal injection of rat
(hot plate incubation period) be (mean value SEM) relatively
| Group | Number of rats | Dosage | Hot plate incubation period (second) | ||||||||
| IU/kg | ?μg/kg | ?nmol/kg | Time after the administration (hour) | ||||||||
| ????0 | ?0.25 | ?0.5 | ??1 | ??3 | ??6 | ??24 | |||||
| Control vehicle | ?15 | 12.51± 0.60 | 14.16± 0.64 | 14.32± 1.00 | 14.32± 1.02 | 13.16± 0.79 | 13.14± 0.74 | 12.85± 1.87 | |||
| Elcatonin | ??9 | ??2.4 | ??0.39 | ??0.12 | 13.54± 1.91 | 19.70± 1.75 | 23.90± 3.12 | 24.68± 1.80 ** | 13.0± 0.7 * | 12.1± 0.5 | 10.8± 0.7 |
| ??10 | ??3.6 | ??0.58 | ??0.18 | 12.92± 1.4 | 21.04± 2.22 ** | 26.03± 3.08 ** | 26.08± 3.69 ** | 23.14± 3.05 ** | 20.37± 3.44 | 15.79± 2.40 | |
| ??9 | ??4.4 | ??0.71 | ??0.22 | 13.68± 1.10 | 21.89± 3.12 * | 26.03± 3.36 ** | 26.20± 2.37 ** | 21.11± 2.18 * | 20.02± 3.26 | 11.78± 1.75 | |
| ??8 | ??5.2 | ??0.84 | ??0.26 | 13.41± 0.92 | 27.89± 3.10 ** | 31.55± 4.17 ** | 25.82± 3.79 ** | 28.81± 3.41 ** | 22.97± 2.60 ** | 12.36± 1.69 | |
| ??9 | ??6.0 | ??0.97 | ??0.30 | 11.38± 0.89 | 25.22± 4.13 ** | 28.91± 3.40 ** | 28.48± 3.35 ** | 26.60± 3.87 ** | 24.69± 2.84 ** | 11.17± 1.80 | |
| Morphine | ??9 | ??- | ??37.5 | ??49.42 | 12.81± 1.23 | 22.08± 2.68 ** | 20.97± 2.56 * | 21.81± 2.46 ** | ±19.97 ±2.05 * | 14.02± 1.24 | 11.33± 1.51 |
| ??9 | ??- | ??75.0 | ??98.84 | 13.76± 1.27 | 22.76± 2.43 ** | 21.86± 2.25 ** | 19.88± 2.04 * | 17.42± 1.93 | 17.66± 1.71 | 12.60± 1.19 | |
| ??9 | ??- | ??125.0 | ??164.73 | 13.28± 1.35 | 19.4± 1.6 ** | 2636± 4.20 ** | 26.83± 3.04 ** | 21.69± 3.62 | 15.86± 3.29 | 13.49± 1.63 | |
For persistent period of analgesic activities relatively, lowest dose level elcatonin and morphine (Fig. 4), median dose elcatonin and morphine (Fig. 5) and high dose elcatonin and morphine (Fig. 6) resist the time-histories comparison of injury feel.Obviously, elcatonin is generally than the active longer duration of morphine analgesia, and is obvious especially when low dosage and median dose.The excipient injection does not produce the hot plate that can detect and changes incubation period.
Body weight changes not obvious, does not observe dead example.Behind the intrathecal injection elcatonin, do not see clinical or dystropy in 24 hour observing time.Short time afterbody (2) and hind leg (1) myoclonus appear in two animals (3.6 and 5.2IU/kg).In these trials, do not see tangible morbid state or clinical and dystropy behind the intrathecal injection elcatonin.
In a word, these results show that elcatonin is when the intrathecal route administration, compare with the morphine sulfate of intrathecal drug delivery, promptly produce strong analgesic activity in the molar concentration that is less than about 1000 times of morphine sulfates, and the analgesic activity longer duration that elcatonin produced, reach back 6 hours of injection at least.
Embodiment 10: the analgesic activities of intravenous injection elcatonin
This research manages to determine analgesia effective dose and the definite dose-response relationship through the elcatonin of intravenous route administration.
Totally 24 of the rats of this research use body weight 320-420g.Each dosage level is with 4-8 rat.Through tail vein injection elcatonin 0.5ml/kg body weight.Used dosage is 10,30,100 and 300IU/kg (being respectively 1.61,4.83,16.1 and 48.3 μ g/kg).With top embodiment 5 described hot plates analgesia determination test establishment of base line hot plates reactions with the reaction when injecting back 15 minutes, 30 minutes, 1 hour, 3 hours, 6 hours and 24 hours.
The result of this research shows, the intravenous injection elcatonin only slightly prolongs hot plate incubation period, and expression only has little analgesic effect (table 3).
Table 3
The analgesic activities (hot plate incubation period) of the single agent intravenous injection of rat elcatonin
(mean value standard error)
| Group | Dosage | Number of rats | Hot plate incubation period (second) | |||||||
| ??IU/kg | μg/kg | Time after the administration (hour) | ||||||||
| ????0 | ?0.25 | ?0.5 | ??1 | ??3 | ??6 | ??24 | ||||
| Elcatonin | ??10.0 | ??1.61 | ??8 | 12.03 ±1.02 | 11.95 ±1.40 | 15.64 ±2.39 | 14.95± 2.92 | 14.11± 0.94 | 10.45± 1.03 | 8.86± 1.23 |
| ??30.0 | ??4.83 | ??8 | 10.71 ±1.02 | 10.04 ±0.96 | 12.18 ±1.22 | 14.14 *±1.02 | 14.26± 1.39 | 15.12± 2.20 | 10.76 ±1.25 | |
| ??100.0 | ??16.1 | ??4 | 13.53 ±1.63 | 11.05 ±1.99 | ?9.63 ±1.21 | 11.20± 1.95 | 14.76± 1.19 | 14.00± 1.83 | 14.30 ±1.33 | |
| ??300.0 | ??48.3 | ??4 | 11.22 ±0.38 | 11.05 ±3.82 | 11.65 ±2.31 | 12.65± 2.09 | 17.37± 4.25 | 11.55± 2.80 | 10.52 ±1.74 | |
Higher dosage (100 with 300IU/kg) with compare than low dosage (10 with 30IU/kg), do not see that tangible analgesic effect strengthens, evening (3-6 hour) occurs and slightly prolong though 30-300IU/kg dosage and 10IU/kg dosage are compared the peak effect in the time of 30 minutes.Rat intravenous injection 10-300IU/kg body weight elcatonin is not seen tangible dose-response relationship (table 3).
The preclinical comparative analysis of hot plate of gained shows behind intravenous injection and the intrathecal injection elcatonin, in the time of 0.5 hour, quiet notes lowest dose level elcatonin (10.0IU/kg or 1.61 μ g/kg) produces and gives elcatonin 1.6IU/kg (0.25 μ g/kg) similarly hot plate incubation period (seeing embodiment 8) through intrathecal route.
In these trials, do not see behind the intravenous injection elcatonin that tangible body weight changes, morbidity is routine, dead routine or clinical and dystropy.
Embodiment 11: the analgesic activities of dosage in the elcatonin sheath repeatedly
This research assessment repeated doses intrathecal injection elcatonin is to the analgesic effectiveness.Implant the sheath interpolation pipe to rat as mentioned above, this after the 7-10 angel its from operation process, recover fully.
18 rats are divided into 3 groups, 6 every group.Research is carried out 2 kinds of processing with 1 weekly interval.Processed group I rats received no treatment.II group rat is accepted intrathecal injection sterile saline every day (excipient contrast), continuous 5 days.III group rat is accepted elcatonin (4.4IU/kg body weight), continuous 5 days every day.
Every group by above embodiment 5 is described, measures hot plate incubation period in back 1 hour, 3 hours and 24 hours in 0 minute (before the administration, baseline), the injection of research the 1st day, the 3rd day and the 5th day.
The result of Fig. 7 shows, repeats in the sheath dosage elcatonin and can prolong hot plate incubation period, and back 24 hours of each injection continuously, hot plate was longer incubation period, have through the time accumulation analgesic effect.In these trials, do not see behind the intrathecal injection elcatonin that tangible body weight changes, morbidity is routine, dead routine or clinical and dystropy.
Embodiment 12: the analgesic activities that rat intrathecal injection every day list agent elcatonin lasts 11 days
This research assessment gives the effectiveness of repeated doses elcatonin to rat through intrathecal route, and dosage regimen is for treating interim administration every day, totally 10 continuous single agent in 5 days not treating two of interval at interval in 48 hours.
Implant the sheath interpolation pipe to rat as mentioned above, this after the 7-10 angel its from operation process, recover fully.9 rats are divided into 3 groups, 3 every group.Research is not carried out not treat two 5 days treatment phases of interval in 48 hours at interval.Processed group I rats received no treatment (false injection).In each treatment phase, II group rat is accepted 5 successive intrathecal injection dosage sterile salines every day (excipient contrast), and totally 10 injections are handled in each research.In each treatment phase, III group rat is accepted 5 successive intrathecal injection dosage elcatonins every day (4.4IU/kg body weight), and each research is handled and injected 10 times (table 4) altogether.
Every group by above embodiment 5 is described, at research the 1st, 3,5,7,9 and 11 day 0 minute (before the administration, baseline), injection back 1 hour, 3 hours and 24 hours mensuration hot plates incubation period (table 4).
The result of Fig. 8 shows, repeat in the sheath dosage elcatonin can prolong hot plate incubation period and have through the time accumulation analgesic effect.
Table 4
Research design
| Program | Research day | |||||||||||
| ??1 | ??2 | ??3 | ??4 | ??5 | ??6 | ??7 | ??8 | ??9 | ??10 | ??11 | ??12 | |
| The 1st phase | ||||||||||||
| Injection | ????× | ????× | ????× | ????× | ????× | |||||||
| Hot plate test | ????× | ????× | ????× | |||||||||
| Metabolism is observed | ????× | ????× | ????× | ????× | ????× | ????× | ||||||
| Clinical observation | ????× | ????× | ????× | ????× | ????× | ????× | ||||||
| The 2nd phase | ||||||||||||
| Injection | ????× | ????× | ????× | ????× | ????× | |||||||
| Hot plate test | ????× | ????× | ????× | |||||||||
| Metabolism is observed | ????× | ????× | ????× | ????× | ????× | |||||||
| Clinical observation | ????× | ????× | ????× | ????× | ????× | ????× | ||||||
In these trials, do not see behind the intrathecal injection elcatonin that tangible body weight changes, morbidity is routine, dead routine or clinical and dystropy.
Embodiment 13: the pharmaceutical composition that contains the human serum albumin
This research assessment is as the effect that contains the human serum albumin of elcatonin analgesic composition composition.Human serum albumin (HSA) is to preventing before patient or the laboratory animal administration that elcatonin from adhering to the candidate of medicine bottle or pipe etc.
In each research day, (0.3%NaCl solution, USP) standard 25% (weight/volume) solution of dilution HSA obtain required 0.2% (weight/volume) deposit concentration with normal saline under aseptic condition.Portion with elcatonin solution when needing mixes with equal-volume 0.2%HSA, obtains that the HSA final concentration is 0.1% in the elcatonin compositions in to be injected containing.
As mentioned above, obtain 32 male rats (320-420g), make its adaptation, carry out intubate and make its recovery.I group rat is with 10 μ l/kg volume intrathecal injection 4.4IU/kg dosage elcatonins (in normal saline).II group rat is with 10 μ l/kg volume intrathecal injection 4.4IU/kg dosage elcatonins (in containing the normal saline of 0.1%HSA).The injection of III group rat contains the normal saline 10 μ l/kg of 0.1%HSA.After each administration all with 8 μ l normal saline flushing intubate.
In each research week, behind 0 minute (baseline is before the administration) of the 1st, 3 and 5 researchs day, intrathecal injection, measured hot plate incubation period in 1 hour, 3 hours, 6 hours and 24 hours.Fig. 9 result showed, repeated doses intrathecal injection elcatonin (4.4IU/kg is in 0.1%HSA) back rat hot plate prolongation of latency in 5 days.
In these trials, do not see behind the intrathecal injection elcatonin that tangible body weight changes, morbidity is routine, dead routine or clinical and dystropy.
Embodiment 14: rat vein and intrathecal injection
125Pharmacokinetics behind the I-elcatonin
The design of describing according to table 5 is studied pharmacokinetics/material balance and in selected in-house distribution on 21 Sprague-Dawley rats.
With radioiodinated elcatonin (
125The I-elcatonin) makes the intubate that intravenous injection (IV) and underwent operative insert through jugular vein and make intrathecal injection (IT).After single agent 1 μ g/kgIT bolus dose or single agent 50 μ g/kg IV bolus dose, gather blood, blood plasma, urine, excrement, tissue and corpse sample in official hour (table 5).
Table 5
Research design and sampling time table
| Animal groups (n) | Route of administration | Dosage | Be used for the biological sample that gross activity is measured | Sampling time |
| ?1 (n＝4) | IV (injecting) | Single agent: 50 μ g/kg 125I-elcatonin (20 μ ci/ μ g) volume injected: 1mL/kg | 8 plasma samples of each animal are gathered brain, spinal cord, lung, liver, thyroid, kidney, blood and plasma sample (totally 34 parts of blood plasma, 2 parts of blood and 12 parts of tissue samples) from 2 animals | Behind the blood dosing 5 minutes, 0.5,1,2,4, after tissue, blood and the blood plasma administration in 8,24 and 48 hours 120 hours (n=2 rat) |
| ?2 (n＝4) | In the sheath | Single agent: 1 μ g/kg 125I-elcatonin (20 μ ci/ μ g) volume injected: 20 μ L/kg | 8 plasma samples of each animal are gathered brain, spinal cord, lung, liver, thyroid, kidney, blood and plasma sample (totally 34 parts of blood plasma, 2 parts of blood and 12 parts of tissue samples) from 2 animals | Behind the blood dosing 0.5,1,2,4,6, after tissue, blood and the blood plasma administration in 8,24 and 48 hours 120 hours (n=2 rat) |
| ?3 (n＝4) | IV (injecting) | Single agent: 50 μ g/kg 125I-elcatonin (20 μ ci/ μ g) volume injected: 1mL/kg | 7 parts of urine of each animal and 1 corpse sample of 5 parts of each animals of excrement sample (4 corpse samples, 28 parts of urine samples and 20 parts of excrement samples altogether) are washed cage water | 0-6 after the urine administration, 6-12,12-24,24-48,48-72,0-24 after the excrement administration in 72-96 and 96-120 hour, 24-48,48-72, and corpse administration sky was 120 hours in 72-96 and 96-120 hour |
| ????4 (n＝4) | IV (injecting) | Single agent: 50 μ g/kg 125I-elcatonin (20 μ ci/ μ g) volume injected: 1mL/kg | Each animal is got brain, spinal cord, lung, liver, thyroid, kidney, blood and plasma sample (totally 24 parts of tissues, 4 parts of blood and 4 parts of plasma samples) | 24 hours (n=2 rats after 1 hour (n=2 rat) and the administration after tissue, blood and the blood plasma administration |
| (n＝4) | In the sheath | Single agent: 1 μ g/kg 125I-elcatonin (20 μ ci/ μ g) volume injected: 20 μ L/kg | Each animal is got brain, spinal cord, lung, liver, thyroid, kidney, blood and plasma sample (totally 24 parts of tissues, 4 parts of blood and 4 parts of plasma samples) | 24 hours (n=2 rats after 1 hour (n=2 rat) and the administration after tissue, blood and the blood plasma administration |
| ????6 ??(n＝1) | Not administration | Do not have | Urine, excrement, blood plasma, tissue, this sample of corpse and wash cage water | Collect sample in 24 hours and be used for context analyzer |
Measure gross activity with γ-calculating instrument.Assess protein bound radioactivity in the blood plasma by measuring with the radioactivity in the plasma protein behind the trichloroacetic acid precipitation.
(the free I of gross activity in the blood plasma
2And iodination of protein) t1/2 (terminal phase half-life) is 29 hours behind intravenously administrable, is 32 hours behind the intrathecal drug delivery.The precipitable radioactive t1/2 of TCA similar (being estimated as 40 hours) behind intravenously administrable and the intrathecal drug delivery.
Behind the intrathecal drug delivery, the deutero-radioactivity of medicine at a good pace is distributed in the tissue and (reached Cmax C in 1 hour
Max).Putting in order of concentration is thyroid＞spinal cord＞brain＞kidney＞blood＞lung＞liver.Do not wish that by any specific theory the radioactivity of thyroid middle and high concentration as if having the result of free I2 in the preparation that gives.Intrathecal injection hindbrain blood plasma radioactive level is after intravenous injection, and expression is difficult to pass through blood brain barrier.86% dosage is recovered after the IV administration.Throw and radioactive about 46% in initial 24 hours, be recovered.The main path of eliminating is to discharge (71%) through urine.
Sequence table (1) general information: (i) applicant: Noble, John F.
Abajian, Henry B. is denomination of invention (ii): the method for treatment pain: (iii) sequence number: 5 (iv) addresses:
(A)ADDRESSEE：Greenlee，Winner?and?Sullivan，P.C.
(B)STREET：5370?Manhattan?Circle，Suite?201
(C)CITY：Boulder
(D)STATE：Colorado
(E)COUNTRY：US
(F) ZIP:80303 (v) computer-reader form:
(A) tool types: floppy disk
(B) computer: compatible type IBM PC
(C) operating system: PC-DOS/MS-DOS
(D) software: PatentIn Release#1.0, version #1.30 (vi) the application's data:
(A) application number: WO
(B) applying date: 18-APR-1996
(C) application materials formerly CLASSIFICATION:(vii)
(A) application number: US 08/424,866
(B) applying date: 18-APR-1995
(viii) agent's information:
(A) name: Ferber, Donna M.
(B) number of registration: 33,878
(C) case number: 1-95A WO
(ix) telecom information
(A) phone: (303) 499-8080
(B) fax: the information of (303) 499-80892.SEQ ID NO:1:
(i) sequence signature:
(A) length: 32 aminoacid
(B) type: aminoacid
(C) chain: irrelevant
(D) topological structure: the unknown
(ii) molecule type: polypeptide
(iii) suppose: not
(iv) primary source:
(A) organism: eel
(xi) sequence description: SEQ ID NO:1:Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu1 5 10 15His Lys Leu Gln Thr Tyr Pro Arg Thr Asp Val Gly Ala Gly Thr Pro
The information of 20 25 302.SEQ ID NO:2:
(i) sequence signature:
(A) length: 33 aminoacid
(B) type: aminoacid
(C) chain: irrelevant
(D) topological structure: the unknown is molecule type (ii): polypeptide is (iii) supposed: (iv) primary source not:
(A) organism: people (xi) sequence description: SEQ ID NO:2:Cys Gly Asn Leu Ser Thr Cys Val Met Leu Gly Thr Tyr Thr Gln Asp1 5 10 15Phe Asn Lys Phe His Thr Phe Pro Gln Thr Ala Ile Gly Val Gly Ala
The information of 20 25 30Pro2.SEQ ID NO:3:
(i) sequence signature:
(A) length: 32 aminoacid
(B) type: aminoacid
(C) chain: irrelevant
(D) topological structure: the unknown
(ii) molecule type: polypeptide
(iii) suppose: not
(iv) primary source:
(A) organism: pig (xi) sequence description: SEQ ID NO:3:Cys Ser Asn Leu Ser Thr Cys Val Leu Ser Ala Tyr Trp Arg Asn Leu1 5 10 15Asn Asn Phe His Arg Phe Ser Gly Met Gly Phe Gly Pro Glu Thr Pro
The information of 20 25 302.SEQ ID NO:4: (i) sequence signature:
(A) length: 32 aminoacid
(B) type: aminoacid
(C) chain: irrelevant
(D) topological structure: the unknown is molecule type (ii): polypeptide is (iii) supposed: (iv) primary source not:
(A) organism: salmon (xi) sequence description: SEQ ID NO:4:Cys Ser Asn Leu Ser Thr Cys Val Leu Gly Lys Leu Ser Gln Glu Leu1 5 10 15His Lys Leu Gln Thr Tyr Pro Arg Thr Asn Thr Gly Ser Gly Thr Pro
The information of 20 25 302.SEQ ID NO:5: (i) sequence signature:
(A) length: 32 aminoacid
(B) type: aminoacid
(C) chain: irrelevant
(D) topological structure: the unknown is molecule type (ii): polypeptide is (iii) supposed: (iv) primary source not:
(A) organism: rat (xi) sequence description: SEQ ID NO:5:Cys Gly Asn Leu Ser Thr Cys Met Leu Gly Thr Tyr Thr Gln Asp Leu1 5 10 15Asn Lys Phe His Thr Phe Pro Gln Thr Ser Ile Gly Val Gly Ala Pro
20??????????????????25??????????????????30
Claims (9)
1. the elcatonin of treatment effective dose and pharmaceutically acceptable excipient are used for intrathecal drug delivery with the application on the medicine of treatment people's intractable pain in preparation.
2. application as claimed in claim 1, the throwing of wherein said medicine is that the about 0.2IU of every kg body weight is to about 15IU with the dosage that makes elcatonin.
3. application as claimed in claim 1, wherein said medicine is mixed with the normal volume of intrathecal injection.
4. application as claimed in claim 1, wherein said pharmaceutically acceptable excipient is the sterilization isotonic saline solution.
5. application as claimed in claim 1, wherein intractable pain be cancer with.
6. application as claimed in claim 1, wherein said medicine is injected or is repeated to inject through the intrathecal route administration by single agent.
7. application as claimed in claim 1, wherein said medicine are pressed about 24-48 hour interval through the intrathecal route administration.
8. application as claimed in claim 1, wherein said medicine by continuous infusion through intrathecal drug delivery.
9. application as claimed in claim 1, wherein said medicine further comprises the human serum albumin.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN96194784A CN1187773A (en) | 1995-04-18 | 1996-04-18 | Method for treatment of pain |
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US08/424,866 | 1995-04-18 | ||
| CN96194784A CN1187773A (en) | 1995-04-18 | 1996-04-18 | Method for treatment of pain |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1187773A true CN1187773A (en) | 1998-07-15 |
Family
ID=5128822
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN96194784A Pending CN1187773A (en) | 1995-04-18 | 1996-04-18 | Method for treatment of pain |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1187773A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107921067A (en) * | 2015-07-10 | 2018-04-17 | 杜克大学 | Method for treating pain |
| CN113230385A (en) * | 2021-05-08 | 2021-08-10 | 深圳市保尔医疗服务有限公司 | Pharmaceutical formulation for relieving cancer pain and cancer and use method thereof |
-
1996
- 1996-04-18 CN CN96194784A patent/CN1187773A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107921067A (en) * | 2015-07-10 | 2018-04-17 | 杜克大学 | Method for treating pain |
| US11389483B2 (en) | 2015-07-10 | 2022-07-19 | Duke University | Methods for treating pain |
| CN113230385A (en) * | 2021-05-08 | 2021-08-10 | 深圳市保尔医疗服务有限公司 | Pharmaceutical formulation for relieving cancer pain and cancer and use method thereof |
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