TWI300717B - Novel use of botulinum toxin for the treatment of neoplasm - Google Patents

Description

1300717 A7 B7

This patent application is part of the continuation application of U.S. Patent No. 0 9/4 5 4,8 42, which is filed on the 7th of November, 1999. BACKGROUND OF THE INVENTION This invention relates to methods of treating neoplasms. In particular, the present invention relates to a method for treating secretory neoplasms (whether benign or malignant) and pheochromocytic cells by topical administration of neurotoxins. Adrenal medulla The adrenal gland is a small triangular tissue located at the tip of the kidney. Each adrenal gland includes the adrenal cortex (or outer layer) and the adrenal medulla (or inner layer). Cortical circumference = and surrounded by the medulla. ' . % Adrenal cortex secretes hormones such as hydrogen-based corticosterone and aldosterone. Hydrocorticosterone is produced when it is stressed, regulates sugar utilization and is necessary to maintain normal blood pressure. Aldosterone is the main blender of salt, potassium and water balance. Hydrogen-based corticosterone and aldosterone replacement therapy are necessary if both adrenal glands are removed. Adrenal medulla secretes catecholamine adrenaline (e.g., adrenaline) and noradrenalin (norepinealin). These hormones are important for the normal regulation of various body and body functions, including elevated blood pressure, cardiac withdrawal, and blood glucose concentrations during an urgent response. Removal of the adrenal medulla only results in a slight or no hormonal deficiency, which can be compensated for by other glands in the body. Conversely, excessive catecholamine production can be fatal. In normal adult males, about 85% of the total catechins produced by the adrenal medulla are adrenaline, and the remaining 15% are norepinephrine. The gramme of this medulla is applicable to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). 1300717 A7 B7 V. Description of the invention (. The tissue contains about 1.6 mg of catecholamine. Most of it is found in blood and urine. The medulla is not derived from the adrenal gland, but from the end of the sympathetic ganglia. If the fresh adrenal gland is placed in a fixative containing potassium dichromate, the medulla will turn brown. The reaction is called chromate reaction and indicates the affinity of the adrenal medulla tissue to the chromium salt. Therefore, the adrenal medulla cells are often called chromaffin cells. The collateral cells are also outside the adrenal medulla, but usually only secrete norepinephrine. , rather than adrenaline. Adrenal medulla can be regarded as a sympathetic nerve node innervated by preganglionic biliary stimulating nerve fibers. These nerve fibers can release acetylcholine and excretion of adrenal medullary chromaffin cells. The mode of secretion of catecholamines (mainly adrenaline). The normal adrenal medulla is dominated by the visceral 4 sacral (the sympathetic nervous system preganglionic biliary stimulating branch). Adrenal medulla activity is almost complete It is controlled entirely by this type of biliary stimulating nerve. The pheochromocytoma chromaffin cells (including adrenal medullary chromaffin cells) and sympathetic ganglion cells have many things in common, since they all originate from the same embryonic ancestor. That is, the sympathagonium of the nerve ridge (shown below). The various types of tumors that can be produced by various cell types are represented by brackets. Each of the listed cell types can secrete catechins. Neural ridges > 1 sympathetic germ cells (sympathagonium) (sympathagonioma-malignant) -5- This paper scale applies to Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1300717

V. Invention description G) chromoblastic sympathetic neuroblasts (chromoblastoma or malignant dark pigment (neuroblastoma, often malignant, phaebchromocytoma) found in children) collateral ganglion Cells (dark pigmented cytoma or pheochromocytoma) (ganglioblastoma _ benign) Most pheochromocytoma occur in the adrenal medulla, while atopic and multiple stromal tumors are known to be most common in children. . 1. Paraganglioma The accessory nerve (ie, chromosome) can be found in the heart (near aorta), kidney, liver, gonads, and other places, and is composed of chromaffin cells; these chromaffin cells are clearly sourced. From the nerve ridge cells and moving to tightly bind to the autonomic nervous system, the cells. The paraganglioma is a tumor composed of chromaffin fine f derived from the accessory ganglion. Carotid paraganglioma is called carotid paraganglioma and adrenal paraganglioma is called pheochromocytoma or pheochromocytoma. Carotid body is a circular structure found in the adventitia of the common carotid artery. It can be located after the branching blood vessels: color, brown τ soil ugly with ayer's ligament (ayer'sligament), mainly from the external carotid artery. The normal carotid body is 3 to 5 mm in diameter. The transmission of the human nerve should be = the pharyngeal nerve (the ninth pair of cranial nerves). The glossopharyngeal nerve provides motor nerve fibers. The styloid sphincter muscles, the parasympathetic nerves secrete the fibers to the ear 'one king + lower gland and the sensation of the nerve fibers to the tympanic, soft palate and tonsils." In terms of cytology, the carotid body contains a rich cytoplasmic prostitute, 〆, 八土 11] or an elliptical nucleus. This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) -6 · 1300717

Type I cells (collar cells). The cytoplasm contains dense-core particles apparently used to store and release catecholamines. The normal carotid system is responsible for the changes in the composition of the arterial blood. In general, carotid paraganglioma is a rare tumor, but it is the most common type of head and neck paraganglioma. Most of the treatment options for carotid paraganglioma are surgical resection. However, because of its extreme proximity to important blood vessels and nerves, the risk of surgical injury is high (mainly χ _ for cranial nerve defects and vascular damage) and the mortality is estimated to be approximately 3-9%. The size of the tumor is important, and the incidence of complications is significantly higher in patients with direct control greater than 5 cm. Perioperative alpha and beta-adrenergic blockers (if the carotid paraganglioma secretes catecholamines) can be administered or secondary to vascular k# emboli formation prior to surgery. Radiation therapy (whether applied alone or in combination with surgery) is a more humane consideration and still controversial. Unfortunately, paraganglioma (including carotid paraganglioma) may not be operated due to location and/or size. 2. A collateral cell tumor pheochromocytoma occurs in the adrenal medulla and can cause clinical symptoms associated with excessive production of catechins, including a sudden increase in blood pressure (hypertension), headache, tachycardia, excessive rest Sweating, sudden onset from a bent position, produces symptoms and anxiety. Abdominal development and 24-hour urine test catecholamines are usually sufficient to confirm the diagnosis. Blocking catechol with phen〇xybenzamine and metyrosine usually improves symptoms and prevents the risk of high blood pressure during surgery (selective therapy options). The standard treatment is laparoscopic adrenalectomy, but partial adrenalectomy is commonly used in familial form of pheochromocytoma. Malignant (cancer) pheochromocytoma is rare This paper scale applies to Chinese National Standard (CNS) A4 specification (210X297 public) 1300717 A7 B7 V. Invention (5) tumor. Approximately 0.3% of patients with secondary hypertension are estimated to have pheochromocytoma. Pro-chromoblastoma can be fatal if not diagnosed or treated improperly. Autopsy showed that many chromosomal cell tumors were not clinically suspected and undiagnosed tumors were significantly associated with pathological consequences. Changes in the adrenal medulla can progress from the normal adrenal medulla to the proliferative adrenal medulla (increased number and size of the entire adrenal medulla cells, without specific tumors), and progress to adrenal medullary tumors (pro-chromoblastoma). The pheochromocytoma treatment surgically removes the unilateral or bilateral adrenal glands. Whether to remove the bilateral adrenal glands depends on the degree of disease. Patients who have had bilateral adrenal glands have to take hydrogen corticosterone and aldosterone daily. Hydrogen corticosterone may be replaced by any of hydrocortisone, cortisone or prednisone and must be taken daily. The acid ketone can be replaced by fluorinated ketone ketone (Florineftm) per ounce. Patients need to increase the amount of hydrocorticosterone or prednisone replacement during an urgent period (including fever, colds, influenza, surgery, or anesthesia). 3. Hemanocytoma A glomus tumor (one of the paragangliomas) is usually a benign neoplasm, also derived from the neuroectodermal tissue, which can be found in various parts of the body. Hemangioma is the most common benign tumor that occurs in the tibia and is converted to malignant and metastatic less than 5 ° / 〇. Hemangiomas are derived from vascular spheres of parasympathetic nerves distributed along the base, chest, and neck of the skull. Each side of the ear typically has three vascular spheres. The vascular spheroids are often accompanied by Jacobsen's (CN IX) or Arnold's (CNX) nerves or in the outer membrane of the jugular bulb. However, its -8- paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1300717 A7 ι_____ B7 V. Description of invention (6) Physiological position is usually osteophyte mucosa (tympanic glomus tumor (gl 〇muS tympanicums)), or jugular bulb (jugular bulbar tumor), the incidence of jugular bulbar tumors is 1: 1, 3, the population is the most popular, and the epidemiology is the most attractive. For women, women: male incidence rate is at least 4: 1. The rate of catecholamine-releasing (functional) tumors is about 1% to 3% of the cases. 血管 The glomus tumor has the possibility of secreting catecholamines, which are similarly derived from the adrenal medulla of the spinal cord, which also secretes catecholamines. The corresponding tumor of the adrenal medullary hematopoietic tumor is a pheochromocytoma, and the glomus tumor is called an extra-adrenal pheochromocytoma. The glomus tumor that secretes catecholamines can cause irregular heart rhythm, excessive sweating, headache, nausea, and pale face. Hemangiomas can originate from different parts of the base of the skull. When confined to the middle ear region, it is called the epiphysis (tympanic vascular bulb). When present in the jugular foramen, the degree of extension is called jugular bulbar tumor. When it extends from the neck all the way to the carotid hole, it is called a vagal glomus tumor. When initiated from the carotid artery branching region, it is called a carotid body tumor. Other known glomus tumor sites include the larynx, eyelids, nose, and aortic arch. Jugular bulbar tumor is the most common tumor in the middle ear. These tumors are often densely packed with blood vessels and provide nutrients from the external carotid branches. Symptoms of jugular bulbar tumors include hearing loss, dizziness, and sometimes ear pain associated with irritating sounds in the ear. The patient may lose hearing due to obstruction of the middle ear, but hearing loss may also be caused by a tumor mass that damages the nerve. Cerebral nerve itch that controls swallowing, nausea, shrug and tongue activity may be part of the symptoms of jugular bulbar tumors. Red/blue turbulent masses are common when examining the tympanic membrane. The initial symptoms are concealed. The paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm). V. Description of the invention (7. Due to the location of the tumor and its vascular nature, the most common symptom is inflammatory ear 2 . In most cases, the tinnitus is secondary to the mechanical effects of tumor bulging. Other common symptoms are ear swelling and (conducting) hearing loss. Currently, the blood cell tumor of the amine can be secreted. The treatment consists of first administration and beta blocker followed by 'radiation therapy and/or surgical dissection. Treatment of cephalic bulbar tumors involves administration of alpha & beta blockers. X-ray therapy can be used to improve symptoms (although The block may remain.) It is also possible to use a substance that blocks the blood supply to embolize the tumor. However, this step can lead to problems such as tumor enlargement, which can press the brain stem and cerebellum, and the cells that die due to loss of blood supply will release The tea is tested for amine. Small tumors in appropriate position can be operated on. The complications of cephalic bulbar surgery are tickles in the ear that continuously leak cerebrospinal fluid and control facial activity, feeling and hearing. Even if the operation is successful, jugular bulbar tumor is still a problem, because of its high recurrence rate and may require multiple operations. Surgical dissection has the risk of #39, mainly due to iatrogenic cranial nerve defects and CSF leakage. Lack of cranial nerves Protection may be the most obvious reason for surgical intervention, because it is prone to complications associated with hypocerebral nerve defects. Radiation therapy also has serious complications, including radioactive osteonecrosis of the tibia, brain necrosis, pituitary gland Insufficient function, and secondary malignancy. Other postoperative complications include (10) leakage, respiratory syndrome, meningitis, pneumonia, and wound infection. The anaerobic, gram-positive bacterium, Clostridium faecalis (: 1 〇 汕 bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo bo Mother. Clostridium botulinum spores can be found in soil and can be grown in food cans from home-style canned workers, which are not properly sterilized and sealed, which is caused by many The cause of intestinal cuppox poisoning. Lactobacillus poisoning usually occurs 18 to 36 hours after ingestion of foods infected with Clostridium faecalis culture or spores. Lactobacillus can obviously be lining and attacking through the intestine without attenuation. Peripheral motor nerve 70. The symptoms of sputum toxin poisoning are light, walking, choked, and difficult to speak. 'Severely, the respiratory muscles are itchy and dead. Type A sputum toxin is the most deadly natural biological agent known to man. Enterotoxin (purified neurotoxin complex) has a LD^ of about 5 〇 picograms per mouse. The unit of sputum toxin is defined as one for each weight of 2 gram I Swiss Guardian (Swiss) Webster) ld^ dose of intraperitoneal injection in female rats. Seven immunologically distinct serotonin neurotoxins have been identified, namely, serotypes of serotypes of serotypes: 8, (1, 1), £, 17 and (5; each can be distinguished by type-specific antibody neutralization. Animal species infested by various serotypes of daids toxins and the severity and duration of sputum caused by them are different. For example, it has been determined that the A-type coliform toxin is 500 times stronger than the B-type faecal toxin in terms of the speed of groin. In addition, it has been determined that for the long-term class, the 480 unit/kg dose of the type 3 faecal toxin is still non-toxic, and this dose is about the type A sputum toxin to the primate [仏❹. The sputum toxin apparently binds to biliary motility motoneurons with high affinity and is transferred into neurons and blocks the release of acetylcholine. The sputum toxin has been used in clinical combinations for the treatment of neuromuscular disorders characterized by skeletal muscle hyperactivity. Type A sputum toxin has been approved for use in the United States by the Food and Drug Administration. -11 - This paper scale applies the SS standard (CNS) A4 specification (21GX297 public)~ ----- 1300717 A7 B7 V. INSTRUCTIONS (9) Eyelids, strabismus and half-faced sputum. The non-Α-type sputum toxin serotype is significantly less potent and/or shorter in activity than the sputum-type sputum toxin. The clinical effect of injection of type A sputum toxin into the distal muscle is usually seen within one week after injection. The typical symptom reduction of a single intramuscular injection of a toxin of type A is approximately 3 months. Although all of the serotypes of the sputum toxin can significantly inhibit the release of neurotransmitter acetylcholine from neuromuscular binding, they each affect different neurosecretory proteins and/or divide these proteins at different sites. For example, both type A and E-type faecal bacteria divide the 2,500 keloid (kilDalton, kD) synaptosome associated protein (SNAP-25), but both calibrate different amino acid sequences in the protein. The serotypes B, D, F, and G act on small sac-associated proteins (VAMP, also known as synaptobrevin), which cleave different parts of the serotype. Finally, the daricin toxin type C i was shown to simultaneously cleave syntaxin and SNAP-25. These differences in mechanism of action can affect the relative potency and/or duration of action of various serotypes of the serotype. The molecular weight of all of the four strains of the daricin toxin serotype is about 150 kD. Interestingly, the fusiform toxin complex released by Clostridium includes a 150 kD daricin toxin protein molecule and a related non-toxin protein. Therefore, Clostridium can produce A-type faecal toxin complexes of the type 900 kD, 500 kD and 300 kD. The toxin-like type of sputum is obviously only one size of 500 kD complex. Type D-rhinoxin has two complexes of 300 kD and 500 kD. Finally, there is only one complex of about 300 kD for E and F-type faecal toxins. These complexes (ie those with a molecular weight above about 150 kD) contain non-toxic hemagglutinin proteins and non-toxins and non-toxic non--12-paper scales applicable to China National Standard (CNS) A4 specification (210 X 297) PCT) 1300717 A7 B7 V. INSTRUCTIONS (10) Hemagglutinin protein. When the toxin is ingested, the two non-toxin proteins, which are a neurotoxin complex associated with the molecular composition of the daunoutoxin, provide stability, avoid denaturation of the daids toxin molecules and protect them against digestive acids. In addition, larger (greater than about 150 kD molecular weight) daricin toxin complexes may result in a slowing of the rate of diffusion of the daunoutoxin from the site of the intramuscular injection of the daricin toxin complex. In vitro studies have shown that the enterotoxin can simultaneously inhibit the release of the brain stem tissue from the original cell culture (caused by potassium cations) acetylcholine and norepinephrine. In addition, it has been reported that scorpion scorpion toxin inhibits the release of glycine red and sulphate from the original culture of spinal cord neurons and that the serotonin toxin inhibits the release of various neurotransmitters from brain synaptophysin culture preparations: Acetylcholine, dopamine, norepinephrine, CGRP and glutamate. Preparation of a type A sputum toxin A culture method in which a C. faecalis culture is established and cultivated in a fermenter by a known method, and the fermentation mixture is collected and purified. All of the serotypes of the sputum toxin are initially synthesized as non-activated single-chain proteins, which must be neuroactive by protease splitting or incision. The strains producing the serotypes A and G of the toxins have an endogenous protease, and thus the serotypes of serotonin and sputum obtained from the bacterial culture are mainly active forms. In contrast, the serotype Cl, D& E of the serotype of the serotype is synthesized from a non-proteolytic strain, and thus the one obtained from the culture is typically in an inactive form. Both proteolytic and non-proteolytic strains can produce B&F serotypes, so that the winner can be activated or inactivated. However, even a proteolytic strain which produces a serotype of B-type serotype of B. can only cleave some of the toxin produced. The actual ratio of the incision to the uncut molecule depends on the length of the incubation period and the culture temperature. F -13 - 1300717 A7 _____ B7 V. Inventive Note (11). Thus, some percentages of any preparation (e.g., a B. sinensis toxin) may not be activated, which may indicate the fact that the B-type faecal toxin is much less potent than the type A sputum toxin. The non-activated daricin toxin molecules contained in the clinical preparations contribute to the overall protein content of the preparation, which increases antigenicity without enhancing clinical efficacy. In addition, it is known that B-type sputum venom is less active during intramuscular injection than the serotonin toxin and is also less potent than the A-type faecal toxin at the same dose concentration. A type of sputum toxin has been used in the following clinical combinations: (1) Each intramuscular injection (multi-site muscle) of about 75-125 units of glass to treat cervical tension is insufficient; (2) each time Intramuscular injection of 5 _1 units of the glass to treat the eyebrow line (forehead wrinkles) (5 units of intramuscular injection into the pyramidal muscle and sputum unit intramuscular injection into each enemy eye muscle; (3) by sphincter Inject about 3 〇 _ 8 〇 units of the glass to the puborectal muscle to treat constipation; (4) by intramuscular injection of about 5 units of the glass to the upper eye and the front of the eye In the lower eyelid, the anterior orbital anterior ocular prosthetic muscle is used to treat the eye and the skin is thin and thin. (5) For the treatment of strabismus, the intravitreal injection of the osmosis between about 丨 5 units enters the extraocular muscle. Depending on the size of the injected muscle and the degree of paralysis required for the muscle (ie the required amount of diopter correction). (6) Intramuscular injection of glass through the five different upper limb flexors to treat upper extremity skinny after stroke, as follows: ) flexor deep muscle: 7.5 units to 30 units-14- Paper scale applicable standards (CNS) A4 size · (2ι- cm) 1300717

(b) Flexor digitorum: 7.5 units to 30 units (c) Scapular wrist muscle: 1 〇 unit to 4 〇 unit (d) 桡 屈 flexor muscle: 15 units to 6 units (4) Peptide biceps: 50 units Up to 200 units. Each of the five designated muscles is injected during the same-treatment period, allowing the patient's upper extremity flexor to receive 90 to 360 units of B. sinensis type A toxin in the same treatment period by intramuscular injection. The success of treating various clinical symptoms has raised interest in other serotypes of daids. Two types of sputum toxin preparations (DySp〇rt®) and b-type and ρ-type fluorocarbons have been prepared (one from white) Chemicals (products) preparations were studied to determine the efficacy, safety, and antigenic efficacy of attenuating local muscles. The preparation of the daricin toxin was injected into the head of the right gastrocnemius muscle (〇5 to 200.0 units/kg) and used The toe abduction scoring test (by 忖abduction scoring assay, DAS) was used to assess the degree of muscle attenuation. The eDw値 was calculated from the dose response curve. The ld50 dose was determined by intramuscular injection in different rats. LD^/EDm was calculated as the therapeutic index. Another group of rats received hindlimb injection of B. sinensis (5.0 to 1 〇. 〇 unit / kg) or b-type faecal toxin (5 〇〇 to 4 〇〇〇 unit / kg), and tested the degree of muscle weakness and increased water Intake, which is expected to cause dry mouth. Antigen efficacy is assessed by intramuscular injection of rabbits once a month (1.5 or 6.53⁄4 μg/kg of type B urinary toxin or 毫15 ng/kg of glass opas) Muscle weakening front of all serotypes The duration of action is dose-related. DAS ED5〇値 (units/kg) is as follows: Bosex: 6.7, Daisuke: 24.7, B-type faecal toxin: 27.0 to 244.0, F-type sausage -15- paper The scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1300717 A7 _ B7 V. Description of the invention (13) Toxin: 4.3. The effect period of the glass is better than the B-type faecal toxin or F-type 躐The enterotoxin is long-lasting. The therapeutic index is as follows: Bosex··1〇5, Daisuke: 6.3, B-type faecal toxin: 3.2. Although B-type faecal toxin weakens muscle function, injection type B The water intake of the sputum toxin was higher than that of the B. sinus group. In 4 months after the injection, 4 of the rabbits (treated at 15 ng/kg) and 4 of the rabbits (with 6.5 gram micrograms) / kg treatment) 4 of the rabbits produced antibodies against the toxin of the sputum-type sputum. In another group of the studies, 9 of the rabbits treated with chlorhexidine showed antibodies against the sputum-like toxin. The DAS results indicate that the relative potency of the type A sputum toxin is equivalent to the F-type faecal toxin, while the F-type faecal toxin It is higher than the B-type faecal toxin. In terms of the action period, the type A sputum toxin is longer than the type 3 faecal toxin, and the type 6 faecal toxin is more active than the type F faecal toxin. As shown by the therapeutic index ,, two The products of the type A sputum toxin preparations (both keith and daisy) were different. An increase in water intake was observed after injection of the B-type faecal toxin in the hind limbs. The behavior proved that the serotype entered the clinic in large quantities. The mice were systemically circulated. The results also showed that the same efficacy as the type A sputum toxin was required, and the dose of the tested serotype was increased. An increase in dose is not conducive to safety. In addition, the antigenicity of type B in rabbits is stronger than that of glass, which may be due to the higher protein content of beta-rhinoxin to achieve an effective dose. Acetaminophen The various types of neurons in the nervous system of the animal are typically capable of releasing only a single type of small molecule neurotransmitter. The neurotransmitter acetylcholine can be secreted by multiple brain neurons in the brain, but it is specifically called the large cones of the motor cortex, several different neurons of the basal ganglia, and the motor gods that govern the skeletal muscles. 16- 1300717 A7 B7 V. INSTRUCTIONS (14) Preganglionic neurons of the ectopic, autonomic nervous system (sympathetic and parasympathetic), postganglionic neurons of the parasympathetic nervous system, and some postganglionic neurons of the sympathetic nervous system Secreted. In essence, since most of the post-ganglionic neurons of the sympathetic nervous system secrete the neurotransmitter and norepinephrine, only the post-ganglionic sympathetic nerve fibers that connect the sweat glands, the pilose muscles, and some blood vessels are biliary stimulating. In most cases, acetylcholine has a stimulating effect. However, acetaminophen is known to have an inhibitory effect on some terminal parasympathetic ends, such as the vagus nerve, which inhibits heart rate. The output signal of the autonomic nervous system is transmitted to the body via the sympathetic or parasympathetic nervous system. The preganglionic neuronal system of the sympathetic nervous system is extended by the anterior sympathetic neuronal cell bodies located in the middle and outer meridian horns of the spinal cord. The preganglionic sympathetic nerve fibers extending from the cell body are in contact with the postganglionic neurons located in the parasympathetic ganglia or the anterior vertebral ganglia. Since the preganglionic neurons of both the sympathetic and parasympathetic nervous systems are biliary stimulating, the application of acetylcholine in the ganglion can simultaneously stimulate the postganglionic neurons of the sympathetic and parasympathetic nerves. Acetylcholine activates two types of receptors, scorpion toxin and nicotine receptors. The scorpion toxin receptor is found in all effector cells that receive post-ganglionic neurons of the parasympathetic nervous system and post-ganglionic biliary-stimulated neuronal stimulation of the sympathetic nervous system. The receptors are found in the synapses between the sympathetic and parasympathetic neurons before and after the ganglion. The nicotine receptor is also present on many skeletal muscle fiber membranes at the nerve junction of the nerve. When small, transparent intracellular vesicles fuse with presynaptic neuronal cell membranes, biliary stimulating neurons release acetylcholine. Many kinds of non-neurons -17- This paper scale applies to Chinese National Standard (CNS) A4 specification (210 X 297 mm) A7 B7 1300717 V. Description of invention (1S) Secretory cells, such as adrenal medulla (and PC12 cell line) And islet cells can each release catecholamines and parathyroid hormones from large dense-nuclear vesicles. The PC 12 cell line is a clonal cell line of murine pheochromocytoma cells, which is widely used in the tissue culture mode of sympathetic adrenal development research. In vitro exogenous toxins can inhibit the release of these two types of compounds from both cells via osmotic cells (via electrodeatinization) or by direct injection of toxin into cells isolated from the nerve. It is known that daricin toxin also blocks the release of neurotransmitter glutamate from cortical synaptophyte cultures. There is therefore a need for effective, non-surgical, non-radiotherapy treatment of neoplasms (e.g., proliferative and/or neoplastic, chromaffin cells that secrete catecholamines, including parasupiva, such as glomus tumors). DESCRIPTION OF THE INVENTION The present invention satisfies this need and provides an effective, non-surgical exfoliation, non-radiotherapy treatment for the treatment of various neoplasms, including paragangliomas, such as glomus tumors. The scope of the present invention encompasses a method of treating a tumor, which is directed to a Zanlu Bureau: administering a sausage between about 10-3 units/kg to 2_unit/kg to reduce the size of the tumor and/or The secretion of the tumor is reduced, thereby treating the tumor. The method of the invention can be directly injected into the tumor itself or by implanting the sausage toxin in the tumor or on the tumor. The method within the scope of the invention can be carried out by locally administering to the tumor a (4) ϋ8 = 〇〇 unit 'kg of the faecal toxin.

MG type of any kind, and preferably A paper size suitable for a @家标准(UNS) M specification X 297 mm) -18- 1300717 A7 B7 V. Description of the invention (type 16 glandular toxin, The clinical efficacy of the type A sputum toxin has been proven and easily obtained. The delivery rate of the carcass toxin is from about 1 unit to about 4 〇, the total unit of 〇〇〇, not per kilogram of disease. Suffering weight unit). In the range of higher doses of daricin toxin (ie 40,000 units), it can be administered by means of a controlled release delivery system (ie implant), whereby a portion of the dose of the sausage toxin reservoir (ie about 10 units of the sputum type) A sputum toxin or about 500 units of a serotype 8 toxin can be controlled via a controlled release delivery system over a period of 3 to 4 months (continuous release delivery system), or a multiphasic controlled release delivery system Release in a repeating cycle (pulsating release delivery system) within approximately 3 to 4 months. The appropriate controlled release delivery system of the present invention, whether continuous or sedating, releasing a therapeutically effective amount of a faecal toxin in the vicinity of the tumor or the surrounding tumor, is disclosed in the patent number of the patent in the same Q9/58725〇 'fNeUrotoxin Implant” and the US patent application Botulinum Toxin Implant applied for on July 2nd, 2nd, the serial number is undecided. In a preferred embodiment of the present invention, the local dosage of the type A sputum toxin to a certain part of the tumor body or the tumor body of the present invention may be between about 1 〇 3 and 3 jin to about 40 units / kg. The eight-type faecal toxin of about 1〇-3 units/kg does not pre-shot to achieve a significant effect, while the type A succulent above 4"/male may cause toxicity or near toxin toxicity. For type B glandular toxins, the present invention is administered topically to the tumor (IV) The dose of enterotoxin is between about 1 () · 3 units / kg to about 2 units / 82. Below about 1 〇 · 3 units/kg of sputum-type sputum toxin is not expected to have two obvious effects' and higher than 2_unit/kg (four) sputum venom -19-

1300717 A7 B7 V. INSTRUCTIONS (17 Toxic doses that are toxic or close to type B toxins. According to the reported intramuscular injection of about 2000 units/kg of b-type faecal toxin preparations, the commodity is close to the primate type B bacterium. The lethal dose of toxin. Meyer κ E. et al. A Comparative Systemic Toxicity Study of Neurobloc in

Adult and Juvenile Cynomolgus Monkeys, Mov. Disord 15 (Suppl 2); 54; 2000. As for (:, 1), £, 17, and (} type of sputum toxin, the dose of sputum to tumor can be determined according to the condition of each patient and should not exceed the dose range of type B toxin. In an embodiment, the amount of the a-type daricin toxin administered as in the disclosed method is from about 10.2 units/kg to about 25 units/kg. Preferably, the sustained release system is administered to the B during a fixed period of time. The dosage of the toxin is 34 to 2 units/kg, because, as described above, it has been reported that less than about 1 unit/kg of the type B. Toxins can be intramuscularly injected into primates without systemic effects. More preferably, the amount of toxins of type A is higher than about 1 〇·1 unit/kg to about 单位5 units/kg. Optimally, type A sausage The dosage of bacterin toxin is between about 单位' unit/male==two kilograms. In many cases, 'intratumoral administration of a type A sputum toxin from about 1 unit = 〇 early position can provide effective and long-term treatment. The effect 'is as described above. More preferably, the unit can be used for toxins (such as type A sputum toxin) The effect is remarkable. In the preferred embodiment of the present invention = effective treatment can be produced in the tissue with the praise of the tumor; two of the detailed methods of the invention are one of the two methods for the human patient - 20- 1300717 A7 Description of the invention (18 ′′ '丨 丨 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约 约This method also includes a method for treating functional neoplasms or functional chromophobias, and the second method is to reduce the secretion of edema by the tumor by administering the neurotoxin to the tumor, thereby using the "functionality," , refers to the secretion of catecholamines, and, in addition, the local drug directly injected neurotoxin into the tumor or its local area, and "赘 = (or its synonymous "tumor") refers to faster growth than normal tissue. Misalignment: tissue, which may be a benign tumor or a malignant tumor (ie, cancer). Systemic routes of administration (such as oral and intravenous administration) are not included in the scope of the present invention. Gangliomas or glomus tumors. A sputum toxin, that is, at least one amino acid of the genus Streptococcus toxin is deleted, modified or substituted compared to the natural sputum toxin. Therefore, the sputum toxin may be a recombinantly produced sputum toxin or A derivative or fragment thereof. The preferred method for treating secretory neoplasm according to the present invention comprises the step of locally administering a therapeutic amount of a darictoxin to a secreted neoplasm of a human patient, thereby reducing tumor secretion. The secretory system refers to the secretion of catecholamines, and the secretory tumor may be a functional paraganglioma. In addition, the functional paraganglioma may be, for example, tympanic glomus tumor, jugular bulbar tumor, vagal glomus tumor, and cervical tumor. The present invention also encompasses a method of improving the physical function of a patient, which comprises the steps of administering a neurotoxin to a functional paraganglioma of a human patient, thereby improving various physical functions such as pain relief and shortening of bed time. Increase walking, more cheerful and more lifestyles. -twenty one ·

1300717 V. Description of invention (19 A7 B7

A further step in the method of treating a patient's secretion in the context of the present invention comprises administering to the patient an effective amount of a darictoxin to reduce secretion, wherein the secretion refers to catecholamine secretion. And the secretion can be endocrine secretion from chromaffin cells. In particular, chromaffin cells can be proliferative and/or hypertonic chromaffin cells and the secretion can be affected by biliary priming. Another method within the scope of the present invention is a method for treating the secretion of biliary stimulating I*, endocrine, and catechol-glutamine cells in a human patient by administering a therapeutically effective amount to a human patient. Type A sputum toxin to reduce secretion. The present invention also encompasses a method of treating glandular glands by administering a ganoxin to a gland to reduce glandular secretion activity, wherein the gland is a catecholamine secretory gland. The gland may be an excessively secreted gland and/or the glandular system is affected by the biliary stimulating nervous system. In addition, the toxin can be administered by injection into the gland or to a localized area of the gland. Another preferred method within the scope of the present invention is a method for treating excessive secretion of catecholamine foot glands, the method comprising the steps of injecting a therapeutically effective amount of a type A flutoxin to a human patient to excessive secretion of catecholamine, biliary stimulating Gland or local glandular area affected by the nervous system to reduce excessive catechin secretion. The invention also encompasses a method of treating an endocrine disorder, the method comprising the step of administering a neurotoxin to the mammal to reduce excessive secretion of the endocrine gland. Finally, the invention encompasses a method of treating a disorder of the adrenal gland, the method comprising the step of administering a neurotoxin to the adrenal gland of the mammal to reduce excessive secretion of the adrenal gland. The scope of the present invention also encompasses a method of treating a tumor, the method comprising the step of locally administering a toxin to the tumor or the tumor, thereby causing the tumor-22- the paper scale gg standard ((10)) A4 Specifications (_21Gχ 297 publicly recovered --

Binding

Line 1300717

DESCRIPTION OF THE INVENTION (2〇 size reduction>. The tumor can be a paraganglioma and the diameter of the tumor can be reduced by about 2% to about 1% after topical administration of the daricin toxin. The invention is based on the discovery that the size and/or secretion activity of various tumors can be reduced by treatment with the toxin. In addition, treatment with daunic toxin in vivo can reduce excessive secretion of chromaffin cells and chromaffin secretion. Active. The target tissue is biliary-induced neuronal distribution or sensitive to high toxin doses, so the proteolytic light chain of the parent can be absorbed by biliary stimulating neurons and #赘赘 tumors (such as chromaffin cells and/or The activity of the standard secretory chromaffin cells. Therefore, the functional paraganglioma, pheochromocytoma and glomus tumors of the biliary stimulating nerve distribution can be locally administered with neurotoxins (such as daids toxin). Treatment. By topical administration, the neurotoxin is administered to the tumor itself to be treated, or to the vicinity of the tumor or to the local area of the tumor. Topical administration includes

Intratumoral injection of neurotoxin. Non-cancer (benign), cancer (malignant) Z and/or hypertonic catecholamine secreted tissue can be treated as described within the scope of the invention. The precursor of pheochromocytoma such as nodular or proliferative proliferation can also be treated by the method of the present invention. Therefore, in the early diagnosis, the injection of the sputum toxin can be used to reduce the proliferative, biliary-exciting innervating chromogranin cells to secrete catecholamines. The Applicant has found that the use of a specific neurotoxin (raffin toxin) for the treatment of paraganglioma with catecholamine-secreting activity can result in great alterations and effects, and thus apparently replaces the current surgery and tumor 2 for this type of tumor. : Line therapy. It is noted that a single dose of daunouto toxin can substantially reduce the tachycardia associated with functional paraganglioma, headache, ancient, rm blood pressure, and its paper scale applicable to the Chinese National Standard (CNS) A4 specification. (210 X 297 mm)

-23· A7 B7 1300717 V. INSTRUCTIONS (21) The tea has an excess of amine test. The route of administration and dosage of daunouistic toxins vary widely, depending on the specific tumor disease being treated and the variables of various patients, including size, weight, age, severity of the disease, and response to treatment. The appropriate route of administration and dosage will usually be determined by the attending physician on a case-by-case basis. This type of decision-making method is routine for those skilled in the art (see, for example, Harrison's Principles of Internal Medicine (1997), Anthony Fauci et al., 14th edition, McGraw Hill). Published) For example, for the treatment of tinnitus caused by cerebral glomus tumors, the type A sputum toxin complex solution can be directly intramuscularly injected into the tumor by endoscopic administration, thereby substantially preventing the toxin from entering the systemic circulation. The particular dosage suitable for administration can be determined by those skilled in the art in accordance with the above factors. The dosage may also be determined by the size of the tumor to be treated or removed, and the type of toxin preparation product. In addition, the effective dose of the sputum required to determine neuroresection of other non-tumor tissues can also be inferred to assess the appropriate dose for human use. Therefore, the amount of toxin injected with type A is proportional to the size of the mass and the degree of activity required to treat the tumor. Usually, a total of about 1 to 2000 units of sputum toxin (such as type A sputum toxin) can be administered per kilogram of the patient's body weight to effectively complete the toxin after administration of the neurotoxin near the tumor body or the tumor. The tumor caused by atrophy. Less than about 〇.〇1 unit/kg of daricin toxin has no obvious effect on functional (ie, catecholamine-secreting) tumors, and higher than 2000 units/kg or 35 units/kg of B or A type of faecium Toxin, which achieves a toxic dose of the particular toxin. Careful placement of the needle and low doses of neurotoxin can prevent the systemic distribution of excess daunoutoxin. Better work-24- This paper scale applies to China National Standard (CNS) A4 specification (210X 297 mm) 1300717 V. Description of invention (22 energy paraganglioma dose range / kg + kg to about 25 units (single f,, (Bouokekexin). The actual administration of sputum toxin 2: == the tumor of the tumor _ block - the degree of activity and the use of the hair in the hair::: The main role of the yin is neuromuscular junction Here, the toxin is used to prevent and release the bilirubin. Therefore, although the toxicity of the bacterium is known, the stimulating, presynaptic, and peripheral motor neurons possess the binding affinity. The bacterium toxin can also bind and translocate into a non-neuronal secretory cell, where the toxin then acts on the secretory vascular membrane to stop the protein by known methods such as Dicer. Because of the intestinal tract: the secretory cell (e.g., chromaffin cells) have low affinity, and it is preferred to shoot pantoxin > king to secretory or glandular tissue to provide higher local toxin concentration. Therefore, the present invention can be applied to treat systemic secretion. (including catecholamine secretion) chromophore And tumors (including secretory tumors with little or no biliary-stimulating nerve distribution). The neurotoxins preferably used in the methods of the present invention are darictoxins, such as serotypes A, B, c, D, E, One of the F or G-type faecal toxins. The preferred sputum toxin is the eight-type faecal toxin, which is highly effective and easy to obtain for humans, and is known to be administered intramuscularly to treat bone and smooth muscle. Disorders. The administration method of the neurotoxin of the present invention for treating cancer can be selected according to various judgment criteria, such as the solubility of the selected neurotoxin toxin and the dose of neurotoxin. The amount of neurotoxin administered can be treated according to a specific treatment, and its severity is 25 - This paper scale applies to Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1300717 A7 B7 V. Description of invention (23) Sexuality and other various patient variables (including size, weight, age, and treatment) The response has changed drastically. For example, the range of impact of the tumor is proportional to the amount of neurotoxin injected, and the number of nerves removed is satisfactory (for most dose ranges) It is proportional to the concentration of neurotoxin injected. The method of determining the appropriate route of administration and dosage is usually determined by the attending physician on a case-by-case basis. Such determination is routine for those skilled in the art (see, for example, Harrison's Principles of Internal Medicine). (1997), edited by Anthony Fossey et al., 14th edition, published by McGherson, Inc.. The scope of the present invention encompasses the use of any neurotoxin, which is used locally as a functional paraganglioma for patients. It has a long-term therapeutic effect. For example, any neurotoxin prepared by the toxin type produced by Clostridium, such as Clostridium botulinum, Clostridium butyricum, and Clostridium utilis can be used or applied to the method of the present invention. In addition, all of the serotypes A, B, C, D, E, F and G of the faecal bacteria may be advantageous for carrying out the invention, but as described above, the type A is the best serotype. The invention can inhibit the atrophy and alleviation of human tumors for 27 months or longer. It is known that catecholamines which are released from the permeabilized adrenal medulla cells can be inhibited by the toxin. Furthermore, it is known that daricin toxin inhibits the release of insulin from cells which are permeable (e.g., via electroporation) to insulin-secreting cells. When exposed to the body, due to the lack of a cell surface sputum toxin receptor, these non-neuronal cell membranes are permeable to promote the entry of the sputum toxin into the cell cytosol. Therefore, B. sinensis toxin can obviously inhibit insulin secretion by lysing Synaptobre vin present in the insulin-secreting cell line HIT-15. Boyd RS et al, The Effect of Botulinum Neurotoxin-B On Insulin Release From a Beta -26- This paper scale applies to Chinese National Standard (CNS) A4 specification (210 x 297 mm) 1300717 A7 ____ B7 Five , invention description (24) "~"

Cell M〇v Disord 10(3): 376 (1995). The inventors' argument is that as long as the light chain of the daricin toxin is translocated into the cell matrix, the daids toxin can block any release from any secretory (ie, neuronal, glandular, secretory, pro-chromic) cell type. Small cystic mediator exocytosis. For example, the intracellular protein SNAP-25 is widely distributed in both neuronal and non-neuronal secretory cells, and the type A sputum toxin is an endoproteinase, and its specific receptor is 25 due to biliary stimulating neurons. Has a receptor with high affinity for daunicus and tetanus toxin (and therefore more sensitive to inhibition of small cystic exocrine secretory compounds than other neurons and other cells), so as the concentration of toxins increases, non Bilirubin-induced sympathetic neurons, chromaffin cells and their b-cell types accumulate daids toxin and show reduced exocytosis. Thus, by practicing the present invention, non-cholinergic neurofibrillary fibers and secretory neoplasms with little or no neurological distribution can be treated with a suitably high concentration of daricin to cause the secretory neoplasm (ie, therapeutic functionality ( Therapeutic atrophy of catecholamine secretory) and proliferative collateral cells. • In normal adrenal medulla, the rate of catecholamine secretion is controlled by stimulation of the activity of the pheochromococcal foot nerve. Contrary to the notion that there is no neuronal distribution of dark pigmented cell tumors and that these tumors are not neurologically controlled when they release catecholamines, there is evidence that the tumor has a biliary stimulating nerve distribution. For example, electron microscopy has shown that nerve contacts with small synaptic vesicles contain catechol-containing vesicle cells. In addition, factors such as emotional agitation, hypotension, or diminished conditions that suddenly accelerate the secretion of catecholamines into the circulation by dark pigmented cell tumors also show the fact that the nervous system can affect secretion. In addition, patients with dark smegmatoma who have not seen abnormalities will cause -27- paper scales to be applied to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). 1300717 Description of invention (25 There is a marked increase in urinary adrenaline, which may result from (&) mechanical action (ie, extrusion of § catechin-containing tumors) (b) sympathetic nervous system reflex activation, where the adrenaline system is increased by the tea test Amine production may have accumulated in the nerve endings of patients with dark pigmented cell tumors and/or (c) activation of existing neural distribution of dark pigmented cell tumors. Furthermore, methods of the present invention may provide improved patient function. Improve the patient's function and improve the quality of life by reducing the factors such as reducing pain, reducing bed time, increasing the health of walking ruts, changing lifestyles and/or normal muscle tone recovery. Hfe, Q〇L) is synonymous. q〇l can be assessed using the known SF12 or sF-36 health measurement scoring steps. s F _ 3 6 is used to assess the patient's body and mental health in eight areas. These include physical functions, role limitations due to physical problems, social functioning, physical pain, general mental health, role limitations due to emotional problems, vitality, and general health perception. As compared to the disclosure of the patient population, as described above, the Applicant has found that topical administration of neurotoxin to human patients with chromatin can achieve surprising effects and long-term efficacy. In its preferred embodiment, The invention is carried out by directly injecting a type A sputum toxin to a local area of a tumor or a tumor. It has been reported that at the junction of the nerve gland, the chemical neurotomy of the genus toxin (such as type A sputum toxin) has A fairly long period of action, ie 27 months compared to 3 months on other sites. The scope of the invention does include: neurotoxins obtained or prepared by bacterial culture, toxin extraction, concentration, preservation, freeze drying and/or reconstitution Retanning and neurotoxins' and (b) modifying or recombining neurotoxins, ie by having -28-

1300717 A7 j----_____ _ B7 V. INSTRUCTIONS (%) Know the chemical/biochemical amino acid tampering method or use one of the known host cell/recombinant to recombinant techniques to A neurotoxin deliberately deleted, modified or substituted with an amino acid sequence and a neurotoxin-derived I or tablet prepared therefrom and comprising a portion having one or more attached chromaffin and tumor cell types Neurotoxins. I. The toxins used in the invention can be cold-dried or hollowed out. The soil type 4 is stored in the scorpion. The daricin toxin can be mixed with a pharmaceutically acceptable excipient, a stabilizer, and/or a carrier (e.g., albumin) prior to lyophilization. Freeze-dried or hollowed out dry material can be reconstituted with saline or water. The following examples are provided to provide a general method of the present invention to those skilled in the art to practice the invention and are not intended to limit the scope of the invention. One- or two-eye, single or dual port middle ear endoscopy can be performed. Accordingly, the anatomy can be observed by transmeasal or transtympanic hard-view mirrors at different angles and through a soft-view mirror through the eustachian tube. There are three endoscopic approaches to the middle ear. (1) the transcranial approach after lifting the tympanic ear canal, (2) the tympanic approach through the tympanic membrane incision, and (3) the non-invasive nature of the natural passage through the eustachian tube. way. Example 1 Examination of middle ear blood with an endoscope ^^ A tympanic chamber can be used to observe the tympanic cavity. Teaching, steerable mirror with a diameter of 〇 8 mm (12,000 pixels, viewing angle, 650 mm in full length, 90° declination, and 25 mm in deflection); Maicon

-29-

1300717 A7 B7 V. INSTRUCTIONS (27) (Manufactured by Micromed Co., Dornbim, Austria) can be used for transsphenoidal endoscopy. The patient's head position is 3 0 ° on the side. The transarticular cannula can be inserted through a tubular catheter placed in the pharyngeal orifice of the eustachian tube via an endoscopic (hard 70° view) guide of the other side nasal passage. After removing the hard view mirror, advance the soft steerable mirror through the tubular catheter to the middle ear. To successfully advance the scope to the middle ear, the appropriate width of the Eustachian canal (ie 1.0 mm wide and 2 mm high) is required. The ear canal or the tympanic chamber mirror can be performed using a hard view mirror. Depending on the method chosen, the outer diameter of the mirror can be 2.3 or 1.9 mm and the angle is zero. , 30°, or 7〇. (Karl Storz, Tuttlingen, and Aesculap). When the ear canal approach is performed, the tympanic cavity can be opened by the endoscope to open the tympanic ear canal so that the scope can enter behind the tympanic cavity below the ankle joint. When the tympanic approach is used, the tympanic membrane can be radially incised depending on the area to be detected between the upper or lower 1/4 circumference or the anterior lower 1/4 circumference. The image can be recorded in the digits provided by the S_VHS image resource # Digi-Still Unit and S-VHS Video Recorder (Sony, Vienna, Austria). The visual range is the angle of the mirror (0., 30., Or 70.) depending on. Hey. The mirror only shows the long protrusions of the anvil and the inner wall (the labyrinth wall). 30. The sight glass provides a large, all-round view. Pointing the mirror upwards extends the field of view to the facial nerve tube. The mirror is pointed down to the bottom of the vortex window (round wind〇w niche), the tympanic sinus is visible after the mirror is pointed, and the snail is visible when the mirror is pointed. 7〇. The sight glass provides a wider field of view of the tympanic cavity. In this view, the upper part is the drum and the mastoid sinus, the lower part is the lower part of the tympanic cavity, the rear is the side treasure and the face crypt, and the front is the tube tympan. -30 - 1300717 A7 V. INSTRUCTIONS (28) ' " ---- And with the help of Qi's hand and turning the tip of the mirror can be used to treat the ear canal? Pass the Eustachian tube gorge. Once the steerable mirror reaches the front tympanum, there are options to proceed in two ways: (1) from the top of the tensor tendon to the drum 1: the epitympanum, and then along the tympanic cavity to the mastoid sinus; or (2) Advancing from the iliac humerus into the middle tympanic cavity and advancing to the anvil and tibia joint, and then selecting (a) advancing from the inner side of the sacral bone and the humerus into the mastoid f or (b) (d) the lateral side of the bone to the drum or ( The sac is advancing from the lateral sinus of the humerus. When the mirror passes through the middle tympanic cavity, it passes through the side wall formed by the entire tympanic membrane and can be inspected throughout. According to the approach, the soft mirror can be easily manipulated through the small bones. Without causing harm. In the following examples, the specific dosage of Bosex is based on the judgment of the attending physician based on various trade-offs and considerations and the amount of coliform toxin that does not produce significant systemic side effects in each case. Example 2 Treatment of catecholamine-secreting glomus tumors Female patients aged 58 years have tachycardia, headache, and elevated catecholamine metabolites in the urine. Adrenal function is normal. Confirmed as benign and functional. Hemangiomas are treated with endoscopic injection of 10 to 1 unit of B. pertussis to tumor mass. Serum catecholamines return to normal within 1-7 days and last for 2 to 24 months. The parasympathetic nerves distributed in the skull base, chest and neck, and glomus tumors derived from vascular spheres can be treated according to this method. Example 3 Carotid paraganglioma for the treatment of catecholamine secretion -31 - This paper scale applies to Chinese national standards ( CNS) A4 specification (210X 297 mm) V. Description of the invention "A 44-year-old male patient with a s, a group of lumps was diagnosed as carotid sacral 2, which is a tumor. The carotid paraganglioma is dark brown to the naked eye. Purple is well surrounded by a thin 2 fiber membrane, located in the non-sensitive neck mass on the anterior side of the sternocleidomastoid and the plane of the hyoid bone. The patient shows symptoms associated with catecholamine production, such as fluctuating hypertension Facial flushing and palpitations. Detecting urinary metanephrines and serum catecholamines. Give perioperative alpha and beta adrenergic blockers. Use neck approaching method to know from 10 units to 15 units. B. chlors is injected into the tumor mass, which is close to or controlled by the tongue and throat nerve. Serum catecholamines return to normal within -7 days and continue to be maintained for the next 2 to 27 months. Example 4 Treatment of proliferative adrenal gland A 62-year-old woman presents symptoms of excessive catecholamines, including tachycardia and high blood pressure. The patient quits bananas, vanilla, coffee, tea, cocoa, chocolate, cola drinks, or drugs (such as tranquilizers and cold nasal sprays) 2) 测量. Measurement of adrenaline and norepinephrine breakdown products in the blood and collecting 24-hour urine to confirm excessive catecholamine secretion. Perform a computed tomography (C Τ or MRI to provide a 3 degree spatial adrenal picture), or A nuclear medicine scan attempts to determine the location and size of the tumor. The results of the biopsy showed a proliferative adrenal medulla that had not yet become cancerous. Directly inject the glass from 1 〇 to 15 〇 to the adrenal medulla by endoscopy. Serum catecholamines returned to normal within 1-7 days and continued for the next 2 to 24 months. Example 5 Treatment of sputum with sputum toxins _ pain -32 - 1300717 A7 ___ ____ B7 _ V. Description of invention (30) A 24-year-old female patient with a history of tachycardia and hypertension. Diagnosed as paraganglioma. The tumor was injected directly into the tumor by X-ray photography. Within 1 to 7 days, tachycardia and hypertension were substantially alleviated and the symptoms of the patient disappeared. X-ray photography and biopsy 3 months after injection did not show any signs of neoplasm. Example 6 Treatment of neoplasms with B-type faecal bacteria A 42-year-old female patient with a history of tachycardia and hypertension. Diagnosed as paraganglioma. The tumor was directly injected with 1500 units of a B-type faecal toxic preparation under X-ray photography. Within 1 to 7 days, the tachycardia and high blood were substantially alleviated and the symptoms of the patient disappeared. X-ray photography and biopsy 3 months after the injection did not show any signs of cancer. Example 7 C-type B. sinensis toxin treatment of a tumor A 58-year-old female patient diagnosed with paraganglioma. Preparation of a c-type sputum toxin preparation of between about 1 〇 3 units/kg to about 35 units/kg (eg, between about 10 units and about 1 〇, a unit of C-type faecal toxin preparation) ()) injected directly into the tumor. Within 1 to 7 days, the symptoms of tachycardia and hypertension were substantially alleviated and the symptoms of the patient disappeared. X-ray photography and biopsy 3 months after the injection did not show any signs of cancer. Example 8 Congwei _^Fungus toxin treatment of blister A 56-year-old obese female patient diagnosed with paraganglioma. Preparation of D-type faecal toxins from about 10·3 units/kg to about 35 units/kg -33 - The paper size is suitable for the wealth of the family X 297 public ^--- 13〇〇717 A7 ---- --- B7 V. Description of the invention (31) ~ ---- The substance (such as between about 10 units to about 丨〇, the unit of D-type sputum toxin preparation) is injected directly into the tumor. During the 7th day, the symptoms of tachycardia and hypertension were alleviated and the symptoms of the patient disappeared. Photogrammetry and biopsy after 3 months after injection did not show any signs of nodules. Example 9 - Treatment of neoplasms with sputum toxins A 61-year-old female patient diagnosed with paraganglioma. A preparation of a sputum-type tocoplasma toxin of between about 1 〇 3 units/kg to about 35 units/kg (eg, a sputum-type tocopherol toxin preparation of between about 1 〇 unit and about 10 〇〇〇 units) Direct injection into the tumor. Within 1 to 7 days, the symptoms of tachycardia and hypertension were substantially alleviated and the symptoms of the patient disappeared. Photographic and biopsy of the sputum did not show any signs of sputum after 3 months after the injection. Example 1 0-34 Lactobacillus remedy 一名 A 52-year-old female patient diagnosed with paraganglioma. Forming a F-type sputum toxin preparation (e.g., between about 10 units and about 10,000 units of the F-type faecal toxin preparation) between about 丨〇3 units/kg to about 35 units/kg directly to the tumor . Within 1 to 7 days, the symptoms of tachycardia and hypertension were substantially alleviated and the symptoms of the patient disappeared. Photographic and biopsy of the sputum did not show any signs of sputum after 3 months after the injection. Example 1 1 〇 腊 腊 毒素 toxin treatment 赘痼 A 14-year-old male patient diagnosed with paraganglioma. a G-type faecal toxin preparation of between about 〇 3 units/kg to about 35 units/kg (eg, -34-

1300717 V. INSTRUCTIONS (Cry) The preparation of G-type sputum toxin from t 1 0 units to about 10 000 units is directly injected into the tumor. Within 1 to 7 days, the symptoms of tachycardia and hypertension were substantially relieved and the symptoms of the child disappeared. Three months after the injection, radiography and biopsy showed no signs of any tumor. The method disclosed herein has many advantages, and the package is as follows: (1) The present invention can avoid many unnecessary operations, and effectively treat functional chromophores, including proliferative, hypertonic and neoplastic catecholamine secretory tissues. . (2) The full administration can be avoided by direct topical application of a neurotoxin such as the present invention. u (3) As described above, the improvement effect of the single topical administration of the neurotoxin of the present invention can last for 2 years or longer. · ° Non-bacterial, however, the present invention has been described in detail with respect to certain preferred methods', but still in the scope of the invention: it is possible to implement the method, the modification, and the modification method. Many types of neurotoxins are useful in the methods of the invention. Further, the local ear administration method of the invention, wherein the diterpene neurotoxicity or the plurality of dauritoxins can be administered simultaneously or continuously. If the a-type adentoxin is administered continuously until the clinical response disappears or the neutralizing antibody is produced, the E-type faecal toxin is administered again. Alternatively, topical administration of A_G type II: a mixture of various toxins to control the desired therapeutic effect, .y month. In addition, the toxin compound can be administered prior to, concurrently with, or after administration of the neurotoxin to provide an additional effect, such as when the neurotoxin (eg, cousin) begins to exert its therapeutic effect, or the neuroresection is more rapidly occurring. The scope also includes the use of neurotoxins (such as daunouto toxin) to treat functional dysfunctional agents (for neurotoxicity of local administration, paper-based e ® home standard (CNS) A4 specifications (2l〇x297 public 35 1300717 A7 B7 V. Description of invention (33) Element). Therefore, the spirit and scope of the following claims should not be limited to the preferred embodiments described above. -36- This paper size is applicable to China National Standard (CNS) A4 specification (210X 297 mm) Ι3007Κ: ^ 本中曰1 Case No. 090118320- Category (The above sheds are filled by this Bureau) Α4 C4 Chinese manual replacement page ( September 1996) issued ^ I --- new patent specification Chinese name English

New use of daunouto toxin for treating tumor 1; iDVEruSE^B〇TULIHUKrrDXIN FOKTHE TREATMENT OF NEOPLASM Name Nationality

Stephen Renovan STEPHEN DONOVAN Canada i Inventor's Residence, Residence, 27252 Calinai Joe Street, Capizentino, California, USA Bind Name (Name) Nationality Ottoman ALLERGAN, INC. US Line III, Applicant Residence, Residence (Company) Representative Name: 2525 Dupont Road, Irvine Township, California, USA

Martin Affitte MARTIN A. VOET This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm)

Claims (1)

I300$l〇y〇ii832 专利 Patent Application Patent Application Scope Replacement [96^^]) Patent Application Fan Park - A pharmaceutical composition for treating sputum, which comprises (10) 2,000 units/kg of faecal toxin. a kg to 2. 6. 9. I, as in the pharmaceutical composition of the first paragraph of the patent range, wherein the "in the W" is between 1 unit and 4 〇, 〇〇〇 unit (total unit). ΠSpecial: The pharmaceutical composition of the first item, in which the tablets of the toxin toxin are in the range of 103 units/kg to 35 units/kg. ', the pharmaceutical combination of the first item of the second patent range The dosage of the waxy fungus is between 10.2 units/kg and 25 units/kg. The pharmaceutical composition of the invention is in the range of 10-2 units/ From kilograms to 15 units/kg. For example, the pharmaceutical group composition of the scope of patent application, i.e., the dosage of the sausage is between 1 unit/kg and 10 units/kg. ” Shen: Patent No. 1 The pharmaceutical composition, wherein the administration method of the daunic toxin is to implant the genus of the genus toxin into the tumor or on the tumor. A pharmaceutical composition according to the scope of the patent application, wherein the tumor is a paraneoplastic tumor. For example, the pharmaceutical composition of the patent scope of the first aspect, wherein the tumor is vascular tumor 0 1 0 · The pharmaceutical composition of the invention patent range i, wherein the faecal fungus is selected from the free toxin 8 , 8 . A group consisting of 1,0, £, 卩 and 0 serotypes. 1 1 The pharmaceutical composition of claim i, wherein the neurotoxin is a toxin of type A. 1 2 · For example, the pharmaceutical composition of the patent scope of the patent, i.e. A8 B8 C8 Sixth, the scope of application for patents 1300717 by the direct injection of sputum toxins for local injection of drugs. 1 5 · The pharmaceutical composition of claim 13 of the patent scope is nodules. Wherein the tumor is an accessory nerve, a pharmaceutical composition for treating a tumor, and reducing the size of the tumor, which comprises 朦' wherein the tumor is an accessory nerve. 17. The pharmaceutical or composition of the patent scope 16 is a tumor. i 8 • A pharmaceutical composition as claimed in claim 13 wherein the tumor scale can be reduced by 20% to 1%. 2. A medical group of humans that treats tumors and reduces the diameter of the tumor by 2% to 丨〇〇%, which includes a therapeutically effective dose of a toxin. 2〇. The use of 10·/unit/kg to 2〇〇〇 unit/kg of daricin toxin is used to prepare a medicament for treating a tumor. 2 i For the purposes of application No. 20 of the patent scope, the dose of the daunic toxin is between 1 unit and 40, and the unit is 总 〇 ( (total unit). 2 2 • For the purpose of claim 20 of the patent scope, the dosage of the daunic toxin is between 10 and 3 units/kg to 35 units/kg. 23•If the application of the scope of patent application is 20, the agent of the sputum toxin is 10 丨 10 10 10 10 / / / / / / / / / / / / 24 • For the purpose of applying for the scope of patent item 20, the agent for daunic toxin 1300717 Patent application park A8 B8 C8 D8 The quantity system is between 10.2 units/kg to 15 units/kg. 25. In the case of the second paragraph of the patent application, the dose of the daunic toxin is between 1 unit/kg and 10 units/kg. 26. The use of the second aspect of the patent application, wherein the drug is administered by implanting the drug implant into the tumor or on the tumor. 2 7 ·If the application of the patent scope of the 20th item, the use of the tumor-type paraneoplastic 〇28·such as the scope of the patent scope of the second paragraph, the towel of the tumor glomus tumor 〇 2 9 · The scope of the patent application scope is the second item*, wherein the faecal strain is selected from the group consisting of serotypes of sputum A 'B, C 丨, D, E, F and G. 〇 · For the purpose of the second paragraph of the patent scope, the neurotoxin is a type A sputum toxin. 31. The use of the second aspect of the patent application, wherein the daricin toxin is administered locally by direct injection of a daricin toxin. 32. Use of a 10-3 unit/kg to 2 〇〇〇 unit/kg type A darictoxin for the preparation of a secretory drug for treating a tumor and reducing a tumor. ' 33. For the purposes of application No. 32 of the patent application, wherein the secretory refers to the (4) amine secretion. 34. For the use of the scope of claim 32, wherein the tumor is a paraganglioma 0 35_ a use of a toxin, which is used for the preparation of a tumor for treatment 8 8 8 8 A BCD 1300717 VI. Scope of application for patents Small tumor size drugs. 3 6 · For the purpose of claim 35, wherein the tumor is a paraganglioma. 3 7 ·If the application of patent application category 32 is used, the diameter of the tumor can be reduced by 20% to 100% 〇3 8 · The therapeutic amount of daric bacteria. Toxin is used for preparation for treatment. It is a tumor that is less than 20% to 100% of the diameter of the tumor. This paper>^ Cancer view 嘉爵丽^ Standard (CNS) A4 specification (210 χ 297 public)