SCORPION VENOMS (For Pharmaceutical Applications) ABOUT US

Satix Pharma Co. began its activity in the pharmaceutical industry in to produce herbal medicines and venom products extracted from insects and ar- thropods.

This brochure highlights some infor- Satix Pharma Co. and its partners The Satix Pharma Co. installations mation about various that are honored to recruit experienced are located in the Fars province in can be found in Iran and the benefits and skilled staff, including toxicolo- Iran being entirely compatible with the of their venom in pharmaceutical ap- gists and laboratory experts who have ecology of the subject in plications. This brochure shows our been working under the supervision terms of climate. ability to breed and raise multiple spe- of the Pharmaceutical and Laboratory cies of and extract, collect Department of the Ministry of Health, Our products are accompanied by and distribute venom of scorpions for so guaranteeing the production of technical information including purity market consumption in accordance to high-quality scorpion venoms. and biological activity from examina- highest quality standards certified by tions certified by state supervised authentic laboratories. laboratories.

2 We will be pleased to host our cus- facility in any Middle East country, tomers who are interested in visiting with more than 3000 square meters the scorpion farm and facilities. Ad- of infrastructure. We are using expe- ditionally, samples are provided for all rienced technicians in this field, and pertinent testing and can be exam- able to produce qualified products. ined by certified laboratories.

Satix Pharma Co. and its partners have legal licenses from the Iranian government to operate the largest scorpion farm and venom extraction

SCORPION VENOMS 3 EXISTING SCORPION VENOMS

PRODUCT NAME ILLUSTRATION/PHOTO

Venom Androctonus Crassicauda

Venom Eupeus

Venom Odontobuthus Doriae

Venom Hottentotta Saulcyi

Venom Hottentotta Zagrosensis

Venom Hemiscorpius Lepturus

Venom Hottentotta Jayakari

4 PRODUCTION FARM Larestan, Iran

SCORPION VENOMS 5 INTRODUCTION

Scorpions constitute a very well adapted order of predatory neurotoxins in the venom. They can block or modify the that have been living in the Earth for nearly 400 mil- functioning of their targeted ion channels in excitable cells, lion years. They now inhabit virtually every terrestrial habitat, which results in autonomic excitation. Scorpion a-toxins except Antarctica. More than 1400 have been de- cause massive endogenous release of catecholamines. scribed to date. The key to their success is the production The combination of sympathetic excitation and the release of potent venoms that they use primarily to kill or paralyze of catecholamines in plasma generates a cascade of physi- their prey and to deter possible competitors and preda- ological events that can progress to arterial hypertension tors. The expansion of human civilization and the growth or hypotension, tachycardia or bradycardia, arrhythmia, un- of human population have led to increased interaction with consciousness, pulmonary edema, heart failure and death. these arthropods, frequently resulting in accidents when Scorpionism represents a major health problem in several people get stung. The immediate pain that the sting elic- countries. More than 1.2 million scorpion stings are regis- its and the devastating consequences that the envenoma- tered globally every year. Despite the fact that only about 25 tion can ultimately cause in human beings attribute to the scorpion species are known to be dangerous to humans, bad reputation of these animals. The effects of a scorpion about 3000 stings per year are fatal. sting can vary widely, from just local pain or inflammation to severe clinical complications, including death. The sever- ity of scorpion envenomation is related to the presence of

TAXONOMY

Scorpion, spiders, ticks and mites are all arthropods. They with thin delicate pedipalps would need extra assistance belong to the , which refers to the structure of from powerful venom. It sounds logical, but it is not known the front appendages. Various authors have proposed dif- if this correct. The color of the is not significant with ferent classification schemes. These are based inter alia on regard to toxicity. The identification of a scorpion at the the form of the sternum, the presence of protuberances species level is a job for specialists. The use of taxonomic on the legs, the morphology of the telson and the number keys presumes a good knowledge of the nomenclature, of lateral eyes. The number of trichobothria (small hair-like the jargon, the morphology and anatomy of the animals. sensory organs) on the femur of the pedipalps is also used. Drawings are generally diagrams which require some inter- Thus the have a triangular central sternal plate. pretation, for which prior knowledge is necessary. Several The other families have a pentagonal sternum. In general, it species are indistinguishable from one another on morpho- can be said that scorpions with massive claws (Scorpioni- logical grounds. These cryptic species can only be distin- dae, Chactidae, Diplocentridae, Bothriuridae) are relatively guished with chemical or genetic methods. harmless, while the more dangerous species (Buthidae, Vaejovidae) have finer claws. An often-heard explanation is that species with massive claws have no difficulty killing their prey, and so do not need powerful venom. Species

6 Families:

• Bothriuridae • Euscorpiidae • Scorpionidae • Buthidae • Hemiscorpiidae • Superstitioniidae • Caraboctonidae • Luridae • Vaejovidae • Chactidae • Microcharmidae • Akravidae • Chaerilidae • Pseudo-chactidae

About only 20-25 types of 1400 species of scorpion world wide are dangerous.

Androctonus: They can make a noise by rubbing their sting over these Thick-tailed scorpions: A. australis, “Androctonus australis” small knobs. They are very venomous. Local pain and sys- is the most important species in the Old World. temic effects and even death can be the result. P. liosoma, “Parabuthus liosoma”, P. granulates, “Parabuthus granula- Buthus: tus”, P. transvaalicus, “Parabuthus transvaalicus”, P. palli- in the case of Buthus occitanus, “Buthus occitanus”, the dus, “Parabuthus pallidus” and P. fulvipes, “Parabuthus ful- major species within this , the venom varies sharply in vipes” are the most notorious species. Parabuthus species toxicity depending on the geographical region. are unique in that they can also spray venom in a fine jet. This probably involves a reflex which is triggered when the Mesobuthus: (earlier placed within the genus Buthotus): animal feels threatened. The animal does not aim the jet in Mesobuthus tumulus, “Mesobuthus tamulus” or “Red a specific direction, but can spray up to 1 meter away and Indian Scorpion” primarily causes cardiovascular problems 50 cm high. (hypertension, arrhythmia, pulmonary oedema). : Hottentotta: This genus includes the most important scorpions in the and H. minox, “Hottentotta minox” are often found under New World. Centruroides suffuses, “Centruroides suffusus” the bark of Acacia trees. The sting is painful. and C. sculpturatus “Centruroides sculpturatus”, and C. exilicauda, “Centruroides exilicauda” are notorious. C. suf- Leiurus: fuses, “Centruroides suffusus” measures up to 90 mm, is Leiurusquinquestriatus, “Leiurusquinquestriatus” is one of yellow to brown in color and has two longitudinal stripes on the most frequently encountered scorpions in the Sahel. the tergites of the pre-abdomen. Although the animal produces relatively little venom, the venom is quite powerful. Tityus: This genus includes more than 100 species, including T. Compsobuthus and Odonthobuthus: serrulatus, “Tityus serrulatus”, (Brazil). This creature is par- C. acuticarinatus, “Compsobuthus acuticarinatus” from thenogenetic. It is fairly common in urban environments and Egypt and O. doriae, “Odonthobuthus doriae” from Iran are – like T. bahiensis, “Tityus bahiensis” – enters houses. T. moderately venomous. trinitatis, “Tityus trinitatis” is highly venomous and is found in sugar cane fields and cocoa plantations. However, this Parabuthus: species is not found in houses. Most species are found in South Africa and Namibia. The animals measure 5-14 cm and are brown-yellow to black. The genus is distinguished from others through stridulatory granules on the dorsal side of the first two tail segments.

SCORPION VENOMS 7 OUR PRODUCTS

1 - Product Name: Venom Androctonus Crassicauda (Arabian fat-tailed scorpion)

Fattail scorpion or fat-tailed scorpion is the common name reading frame was composed of 80 amino acid residues given to scorpions of the genus Androctonus, one of the contain a signal peptide of 22 amino acid residues, followed most dangerous groups of scorpions species in the world. by a mature toxin of 58 amino acids. It had a molecular They are found throughout the semi-arid and arid regions mass of 6.84 kDa and isoelectric point of 4.58. The se- of the Middle East and Africa. They are a moderate sized quence similarity search revealed several matches with the scorpion, attaining lengths of 100 mm (just under 4 in). Their scorpion toxin-like domain of toxin-3 superfamily with a ho- common name is derived from their distinctly fat metaso- mology range of 35-75%. Multiple alignments and second- ma, or tail, while the Latin name originates from Greek and ary structure prediction demonstrated that the toxin peptide means “man killer”. Their venom contains powerful neuro- deduced from the amplified cDNA was related to the long- toxins and is especially potent. Stings from Androctonus chain neurotoxins in size but stabilized by three disulfide species are known to cause several human deaths each bridges instead of four. The level of difference implies that year. Several pharmaceutical companies manufacture an- the corresponding genes have originated from a common tivenom for treatment of Androctonus envenomations. ancestor. This level of difference may also confirm an evo- lutionary link between the ‘short-chain’ and ‘longchain’ Androctonus is widespread in North and West Africa, the toxins. The analysis showed one major segment within this Middle East and eastwards to the Hindukush region. Coun- neurotoxin with maximal hydrophilicity which was predicted tries where Androctonus species live include: , Mo- to be antigenic by inducing an antibody response. rocco, Algeria, Tunisia, Libya, Egypt, Togo, Palestine, Israel, India, Lebanon, , Jordan, Saudi Arabia, Yemen, Despite the risks of keeping such a dangerously venomous Oman, United Arab Emirates, Qatar, Kuwait, , Iran, Af- species in captivity, Androctonus scorpions are frequently ghanistan, Bahrain and . found in the exotic animal trade, A. amoreuxi and A. austra- lis being the most commonly available. The fat-tailed scor- Scorpion venom is the richest source of peptide toxins with pion’s main diet when in captivity consists of , high levels of specific interactions with different ion-channel grasshoppers, and crickets. Scorpions will generally try to membrane proteins. The present study involved the ampli- kill and eat anything which moves and is smaller than them- fication and sequencing of a 310-bp cDNA fragment en- selves. To simulate the desert environment, the enclosure coding a beta-like neurotoxin active on sodium ion-channel used to keep the scorpion in must be kept at a temperature from the venom glands of scorpion Androctonus crassi- of between 26 and 30 degrees Celsius (79–86 °F) cauda belonging to the Buthidae family using reverse tran- scription polymerase chain reaction (RT-PCR) technique. The amplified complementary DNA (cDNA) fragment had a coding sequence of 240 bp. The deduced precursor open-

8 2 - Product Name: Venom Mesobuthus Eupeus

M. Eupeus can reach a size of 40 to 50 millimeters (1.6 intense pain, hyperemia, swelling and a burning sensation to 2.0 in) in length. The entire body is yellow to yellowish in the affected area, while numbness and itching were also brown. The dorsal segments (tergites) of the mesosoma reported in some cases. often have longitudinal irregular stripes that are black to dark brown. They exhibit , the adult fe- As in other scorpions, the venom of M. Eupeus contains males being generally larger than males but have a lower various neurotoxic proteins that interact specifically with ion number of pectinal teeth (16 to 23, as opposed to 22 to 28 channels. A number of unique proteins in this scorpion’s in males). venom have been identified, cloned and investigated for clinical applications. For instance, MeuKTX, structurally The pedipalps of M. Eupeus have a maximum of ten diago- related to BmKTX (α-KTx3.6) from M. martensii, potently nal rows of granules on the fixed finger and eleven on the inhibits rKv1.1, rKv1.2 and hKv1.3 channels but does not movable finger. The pedipalp chelae (pincers) are wider than affect rKv1.4, rKv1.5, hKv3.1, rKv4.3, and hERG channels the patella (segment IV). The segments of the metasoma even at high concentrations. In contrast, BeKm-1 are thick and have eight keels (octocarinate). The telson is specifically inhibits hERG channels, which are potassium subglobose with a flat dorsal surface. channels critical to maintaining normal electrical activities in the heart, but showed no effects on various other potas- Taxonomy: M. Eupeus were first described by the German sium channels tested. Inhibitors of sodium channels have arachnologist Carl Ludwig Koch in 1839. It is classified also been found in this venom. under the genus Mesobuthus and belongs to the largest family of scorpions, the thick-tailed scorpion family Buthi- A number of antimicrobial peptides have also been found in dae. Currently, more than 23 of M. Eupeus are the venom of M. Eupeus. Meucin-13 and Meucin-18 exhib- recognized. ited extensive cytolytic effects on bacteria, fungi and yeasts. Furthermore, Meucin-24 and Meucin-25, first identified from Ecology: M. Eupeus feed on small insects such as crick- genetic sequences expressed in their venom gland, were ets or small cockroaches. Cannibalism is very rare in this shown to selectively kill and inhibit species. They do not dig burrows and prefer using natural the development of , both par- spaces and burrows under stones and other objects. They asites, but do not harm mammalian cells. These two ven- have slender pedipalps so they usually rely on their stings om-derived proteins are therefore attractive candidates for for killing their prey. the development of anti-malarial drugs.

Venom: The venom of M. Eupeus is not as potent as that of other dangerous buthid species. Victims of stings feel

SCORPION VENOMS 9 OUR PRODUCTS

3 - Product Name: Venom Odontobuthus Doriae

Odontobuthus doriae Thorell 1876 (Arachnida: Scorpionida: One of the medically important scorpions is O. Doriae, Buthidae) is a scorpion of medical importance and therefore which exists in relatively high numbers in Iran. LD50, i.e. its geographical distribution in Isfahan Province has been the lethal dose, of this ’s venom is 0.19mg/kg for studied. Methods: This descriptive cross–sectional study mice and thus studying its biological aspects is of value. was designed between March and June in 2012 and 2013 Treatment and preventive measures are important issues in in Province of Isfahan, central Iran. Overall, 164 O. Doriae the field of medicine and hygiene. As such, examining other scorpions were collected from their natural habitat by identi- life aspects of scorpions, including O. Doriae, such as their fying dug out burrows. This arthropod’s burrows were iden- biology and ecology is of importance. tified based on the presence of tumuli, particularly between May and June at the sloping foothills of pristine embank- ments. The sampling data was categorized and compared.

10 4 - Product Name: Venom Hottentotta Saulcyi

Hottentotta is a genus of scorpions belonging to the family They show a typical buthidhabitus with gracile pedipalp- Buthidae. It is distributed widely across Africa. Species also chelae and a moderately thickened metasoma. The vesicle occur in the Middle East, the Arabian Peninsula, southeast- is bulbous and proportionally large in some species. The ern Turkey, Iraq, Iran, , Pakistan, India, Nepal, cephalothorax and mesosoma shows distinct granulation in Cape Verde Islands, Slovenia (rarely), and Sri Lanka (intro- most species, some are strongly hirsute. The tergites of the duced). mesosoma bear three distinct, longitudinal carinae (ridges).

Members of Hottentotta are generally moderately sized As in other buthids the venom in at least some species of scorpions, with a total length of 70–90 millimeters (2.8–3.5 Hottentotta is relatively potent and can be of medical impor- in); the smallest species are 30 millimeters (1.2 in) long, tance to humans. while the largest species reach 100–130 millimeters (3.9– 5.1 in). They belong to the most colorful species of scor- H. tamulus, the Indian red scorpion, has evidently caused pions. The base color ranges from bright yellow to black, human fatalities by cardiovascular and pulmonary mani- with most species colored in brown or reddish tones. Many festations of intoxication and is considered the most lethal species are ornamented by color spots or bands especially scorpion species in the world. on the mesosoma. Coloration may be considerable variable between individuals of the same species or among regional Kleber et al. (1999) listed envenomation by at least four populations. other species as to cause clinically significant symptoms.

SCORPION VENOMS 11 OUR PRODUCTS

5 - Product Name: Venom Hottentotta Zagrosensis

Male holotypes are often 102 mm long, female allotypes part of internal surface. Ventral surfaces of femur and patella usually measure 103 mm long. Trichobothrium on the fixed are smooth to glossy. Patella with 8 carinae. Chela lacks finger of pedipalp located between trichobothria. Males carinae. Movable fingers of pedipalps of this scorpion have possess slightly longer and narrower metasomal segments, 16 rows of granules and 5 terminal granules. Seventh meta- the width of pedipalp chela is the same in both sexes. Pec- somal segment with 4 well marked ventral carinae. First and tinal teeth number 34–36 in males, 27–33 in females. Nearly second metasomal segments with 10 carinae; third seg- entire body is hirsute, pedipalps, dorsal surface of meso- ment with 8 or 10 carinae; fourth segment with 8 carinae; soma, legs, lateral and ventral surfaces of metasomal seg- fifth segment with 5 carinae, 3 ventral (1 median, 2 lateral) ments, and vesicle all densely hirsute. The hairs on patella and 2 dorsal, smooth and sometimes indistinct. Dorsal sur- of pedipalps are long. Color black except reddish brown face smooth and glossy, fifth segment bears 2 short, incon- chela of pedipalps; sometimes ends of first and second tar- spicuous carinae. First metasomal segment of female may someres yellow, coxa and trochanteron ventral side of mes- be wider than long, in the male it is always longer than wide. osoma marbled, and pectens are yellowish brown. Femur of pedipalps with 5 carinae and a row of granules in middle

12 6 - Product Name: Venom Hottentotta Jayakari

Carapace and tergites are usually orange or brown, but uleus is usually more reddish or reddish orange. Pedipalp depending on population can vary from yellowish brown to femur is orange yellow, patella and chela with reddish brown blackish, with eyes surrounded with black, interocular trian- dusky pigment. Legs are yellow to orange yellow or some- gle marked with darker dusky pigment and keels marked times greenish yellow without markings. Ventral surface and with darker brown to black. Tergites often have a slightly sternites are yellowish to orange and paler than dorsum. paler yellowish-brown median strip. Metasomal segments I Pectines are creamish. Chelicerae yellowish to brown with to III are orange yellow (yellowish green in some specimens) dusky reticulations on manus. to orange brown, with segments IV and V often darker. Telson vesicle is orange yellow to orange brown and ac-

7 - Product Name: Venom Hemiscorpius lepturus

Gardium (scientific name: Hemiscorpius lepturus) is the tion of the male is creamy, and the female is brown. It is name of a poisonous scorpion in the Khuzestan prov- rumored that if the victim bites if she does not get paralyzed, ince. The venom of this type of scorpion can easily kill an she will have about 5 minutes to have her antipyretic serum. adult, This scorpion is the most poisonous scorpion in the The severity of muscle contraction is so severe that some- world. Even the Mexican tail scorpion is a distant second. times the victim’s jaw drops out of place. The venom of the Its poison has nothing to do with the killer spider or black scorpion can also destroy the red blood cells. widow poison that lives in the Amazon forests. The colora-

SCORPION VENOMS 13 AVAILABLE VENOMS in our stock:

1 - Venom Androctonus Crassicauda 4 - Venom Hottentotta Saulcyi 2 - Venom Mesobuthus Eupeus 6 - Venom Hottentotta Jayakari 3 - Venom Odontobuthus Doriae

1 2 3 4 6

14 AVAILABLE VENOMS that can be produced:

5 - Venom Hottentotta Zagrosensis 7 - Venom Hemiscorpius Lepturus

5 7

Table - Protein test for venoms

SERIES VENOM SAMPLE TYPES TEST MEASUREMENT UNIT METHOD

Kjeldahl Odontobuthus doriae 90.4 protein g/100 1 method

Kjeldahl Androctonus crassicauda 94.1 protein g/100 2 method

Kjeldahl Hottentotta Saulcyi 91.3 protein g/100 3 method

Kjeldahl Mesobuthus eupeus 95.8 protein g/100 4 method

Kjeldahl Hottentotta zagrosensis 93.1 protein g/100 5 method

The Kjeldahl method was developed in 1883 by a brewer factor (F) is needed to convert the measured nitrogen con- called Johann Kjeldahl. A food is digested with a strong acid centration to a protein concentration. A conversion factor so that it releases nitrogen which can be determined by a of 6.25 (equivalent to 0.16 g nitrogen per gram of protein) suitable titration technique. The amount of protein present is used for many applications, however, this is only an av- is then calculated from the nitrogen concentration of the erage value, and each protein has a different conversion food. The same basic approach is still used today, although factor depending on its amino-acid composition. The Kjel- a number of improvements have been made to speed up dahl method can conveniently be divided into three steps: the process and to obtain more accurate measurements. It digestion, neutralization and titration. is usually considered to be the standard method of deter- mining protein concentration. Because the Kjeldahl method does not measure the protein content directly a conversion

SCORPION VENOMS 15 LABORATORY EQUIPMENT

Venom laboratories requires large variety of equipment and instrumentation to run tests and research.

Our laboratory is equipped to the modern instruments to check the venom quality and activity of several kinds of arthropods and scorpion venoms.

16 LABORATORY EQUIPMENT

Meantime we are using the Pasteur Institute of Iran laboratories that are specialized and equipped in this subject as a well-known and experienced refer- ence source and research center for toxicology and venomous science in Iran.

SCORPION VENOMS 17 CERTIFICATES

Pasteur Institute of Iran is a medical research center lo- Pasteur Institute is a leading regional facility in the develop- cated in Tehran, Iran. The institute is one of the oldest lead- ment and manufacture of vaccines. ing research and public health centers in Iran and the Middle This institute is a center that provides public health services East, was established in 1920 following an agreement be- and has played a significant role in the prevention and tween the Institute Pasteur of Paris and the Iranian govern- control of infectious diseases in Iran and the world during ment. It meets the specialized and scientific health demands approximate one hundred years of its activities of the local community and tries to establish a link between applied research and industry.

18 SCORPION VENOMS 19 SCORPION VENOMS (For Pharmaceutical Applications)

Satix PHARMA Co. Ltd. Unit 2, 152, Zeynali St., North Sohrevardi St., 15769-84418 Tehran-Iran TEL: +98 21 8875 2243 www.satixpharma.com [email protected]

Edition Oct. 2020