CALGEVAX - BCG for immunotherapy

http://www.bulbio.com/media/pdf/en/leaflets/28.pdf

Patient Information Leaflet

CALGEVAX, powder for suspension

(BCG for immunotherapy)

Please read this leaflet carefully before you start taking your medicine because it contains information,

which is important for you.

- Keep this leaflet; you may need to read it again.

- If you have further questions, please ask your doctor or pharmacist.

- This medicine has been prescribed for you personally and you should not pass it on to others. It

may harm them, even if their symptoms are the same as yours.

- If you notice any adverse reactions, please, tell your doctor or pharmacist. This includes all

possible adverse reactions, which are not pointed out in this leaflet. Please, refer to item 4.

In this leaflet:

1.

What CALGEVAX is and what it is used for

2.

What you should know before you take CALGEVAX

3.

How to take CALGEVAX

4.

Possible adverse reactions

5.

Storing CALGEVAX

6.

Content of the pack and additional information

1.

What CALGEVAX is and what it is used for

CALGEVAX is BCG specially designed for cancer immunotherapy.

• Intravesically:

- for prophylaxis of the recurrences of the superficial tumors of the bladder after transurethral

resection;

- for treatment of carcinoma in situ of the bladder.

• Percutaneously:

- in the treatment of malignant melanoma;

- for nonspecific adjuvant immunostimulating therapy of other newly emerged malignant growths

(lung, breast cancer, acute leukemia, lymphosarcoma or osteosarcoma).

2.

What should be known before CALGEVAX is given

Do not administer CALGEVAX if:

-

there is allergy to the active substance or any of the other ingredients of this medicine (listed in

item 6).

Warnings and preventive measures

You should consult your doctor or pharmacyst before using CALGEVAX.

CALGEVAX IS NOT ADMINISTERED:

● in case of patients with immunosuppression;

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● in case of patients with inherent or acquired immune insufficiency caused by a disease or an

antitumor therapy;

● in case of HIV-positive persons and patients on immunosuppression doses of steroids or other

immunosuppresants;

● in febrile conditions;

● in case of infection of the urinary tracts or massive haematuria, as well as in a parallel anti-microbe

therapy;

• in active tuberculosis; a positive tuberculin Mantoux test is a contraindication only in case of

certified active tuberculosis infection;

• In case of ongoing or previous systemic BCG reaction (i.e. a systemic granulomatous disease,

which can be revealed after the administration of BCG; it is determined on the basis of the

following symptoms: fever ≥39.5°С for ≥12 hours; fever ≥38.5°С for ≥48 hours, pneumonitis,

hepatitis or another organ disfunction outside the urinary-genital tract involving a granulomatous

inflammation, which is established through biopsy, or the classical symptoms of sepsis).

NECESSARY PRECAUTIONS FOR USE:

• Patients, who have had the medicine administered intravesically, should maintain adequate

hydration.

• The intravesical administration of CALGEVAX may result in local inflammation of the urinary

bladder, accompanied by haematuria, dysuria, pollakiuria and flu-like symptoms. Patients with a

small capacity of the bladder are exposed to a higher risk of severe local reactions.

• In case of a doubt about a systemic BCG infection after the administration of CALGEVAX, which

may proceed with a fever above 390С, persisting temperature of over 38 0С or fatigue, a prompt

antituberculosis treatment is carried out after consulting a specialist.

• CALGEVAX contains live bacteria and should be treated as infectious material. Persons with

immunosuppression should not handle CALGEVAX.

• The catheterization of the bladder should be performed carefully avoiding traumatization of the

mucous membrane. If the doctor, performing the treatment, decides that the catheterization is

traumatic (for example, involves bleeding), the administration of the product should be postponed

by at least 14 days.

• In case of serious overall adverse reactions, the intervals between the separate administrations are

extended. The treatment with CALGEVAX may be terminated at the discretion of the doctor,

performing the treatment. Antituberculosis treatment is recommended in case of disseminated

BCG infection.

Children and adolescents

CALGEVAX is not recommended for children due to the absence of sufficient data about the safety

and efficiency of the product regarding children.

Other medicines and CALGEVAX:

You should inform your doctor or pharmacyst if you have taken or can possibly take other medicines.

Taking of CALGEVAX may result in an increased sensitivity towards tuberculin, which would

complicate the interpretation of the skin reaction to tuberculin in a future testing for suspected

mycobacterial infections. In this context, it is recommended to determine the patient’s reaction to

tuberculin prior to the immunotherapy with CALGEVAX.

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Pregnancy, lactation and fertility

Do not take this medicine without first consulting your doctor, if you are pregnant or breast-feeding,

think that you might be pregnant or plan to become pregnant.

The risk/benefit balance is assessed carefully, when the medicinal product should be administered

during pregnancy and lactation due to the absence of clinical data about its administration in these

cases. It is known that a lactating woman with a systemic BCG infection may infect her child.

Driving and using machines

The effect of CALGEVAX on the ability to drive and operate machinery has not been studied.

3.

How to administer CALGEVAX

You should take this medicine as described in this leaflet or as your doctor of pharmacyst has told you.

If you are uncertain about something, ask your doctor or pharmacyst.

INTRAVESICAL ADMINISTRATION:

CALGEVAX is administered at least 14 days after the performance of a biopsy, transurethral resection

or traumatic catheterization. The application of CALGEVAX should be done using aseptic equipment

and under the control of a urologist, having experience in the administration of the product. Three or

four ampoules of the product are used for each intravesical instillation at the discretion of the urologist

performing the treatment. The content of each ampoule is reconstituted into 1 ml of sterile saline (a

solution of sodium chloride with concentration of 9 g/l), carefully shaken until obtaining a

homogeneous suspension. The mixture is drawn by syringe and then returned to the ampoule three

times in order to ensure thorough mixing, which minimizes the clumping of the mycobacteria. The

content of the ampoules is transferred in a syringe of 50 ml. A supplementary volume of sterile saline

is added bringing the total volume to 50 ml. Light reduces considerably the efficiency of

CALGEVAX. Therefore, exposure to direct daylight should be avoided both before and after

preparing the suspension. The suspension should be used immediately.

After empting the bladder, the reconstituted CALGEVAX is instilled slowly into the bladder through

urethral catheter. The product remains in the urinary bladder in the course of two hours. While the

CALGEVAX is being retained in the bladder, every 15 minutes the patient alternates the following

positions: semi-left turning, semi-right turning, left side, right side.

The standard scheme of administration includes one intravesical instillation once a week in the course

of six weeks (inducing therapy). The maintenance therapy is determined on a case-by-case basis. The

following schemes can be used: monthly administration in the course of minimum 6-12 months or

three weekly instillations at the 3rd, 6th, 12th, 18th, 24th, 30th and 36th month of the date of the first

instillation.

PERCUTANEOUS ADMINISTRATION:

Ten horizontal and ten vertical lines are made by the syringe needle on an area of 5/5 сm. The

scarification should tear only the epidermal layer without causing extensive bleeding.

A multipuncture apparatus can also be used to regulate the depth of the scarification.

The scarification spot is treated by the following 0.5-ml suspension: the content of one ampoule is

mixed with 0.5 ml of sterile saline to obtain concentration of 75 mg/ml. One ampoule is used for one

scarification. Prior to the procedure, the area should be treated with acetone after which its full

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evaporation should be awaited. A new area is chosen for every following scarification. The frequency

and duration of administration are determined by the doctor, performing the treatment.

4. Possible undesirable reactions:

This medicine, like most other medicines, may cause adverse reaction, although this does not apply to

all people.

Intravesical administration

• Local reactions: transitory dysuria, pollakiuria, haematuria, bacterial infection of the urinary

tract, granulomatous prostatitis, epididymitis, orchitis.

• General reactions: malaise, increased temperature, fever, sweating, sickness, vomiting,

headache, muscle pain, abdominal colic, diffuse rash, liver toxicity, disseminated BCG

infection, generalized hypersensitivity, erythema nodosium, conjunctivitis, uveitis, vitiligo,

arthritis, leucopenia or pancytopenia, splenomegaly.

In case of intravesical administration, adverse reactions (dysuria, pollakiuria, fever) emerge 3-4 hours

after the instillation and are transitory – they last 24-72 hours. A localized (epididymistis, orchitis,

prostatitis) or a systemic BCG infection occurs more rarely.

Percutaneous administration

• Local reactions: localized itching or rash, painful ulcer at the place of administration,

regional adenopathy.

• General reactions: fatigue, increased temperature, fever, sweating, sickness, vomiting,

headache, muscle pain, abdominal colic, diffuse rash, liver toxicity, disseminated BCG

infection, generalized hypersensitivity, erythema nodosium, conjunctivitis, uveitis, vitiligo,

arthritis, leucopenia or pancytopenia, splenomegaly.

In case of serious adverse reactions, the intervals between the separate administrations are extended.

The treatment with CALGEVAX may be terminated at the discretion of the doctor, performing the

treatment. Antituberculosis treatment is recommended in case of disseminated BCG infection.

The patient must inform his doctor of any side effects, not pointed out in the leaflet.

Reporting adverse reactions

If you notice any adverse reactions, please, tell your doctor or pharmacist. This includes all possible

adverse reactions, which are not pointed out in this leaflet. You can also report adverse reactions

directly through the National Reporting System, pointed out below:

Bulgarian Drug Agency

8, Damyan Grouev St., 1303 Sofia,

tel.: + 359 28903417, website: www.bda.bg

By reporting adverse reactions you may contribute to ensuring more information about the safety of

this medicine.

5. Storage of CALGEVAX

Store at a temperature of +2 °C to +8 °С.

Store in a cardboard pack in order to avoid exposure to light.

Keep out of the reach of children.

After resuspending, avoid exposure to light and administer promptly.

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The product should not be used beyond its expiry date, pointed out on the pack.

The product is fit for use until the last day of the month indicated.

Special measures at disposal

Unused amounts of the suspension, needles, syringes and catheters are destroyed in accordance with

the rules for disposal of infectious materials. In case of a spill, the area is cleaned by a 70% solution of

ethyl alcohol.

6. Content of the package and additional information

Content of CALGEVAX

Active substance - Mycobacterium bovis BCG (Bacillus Calmette-Guérin) 11.25 mg (37.5 mg semi-

dry bacterial mass) 1,0 -3,0 x108 viable units

Excipients: Sodium glutamate 40 mg

What CALGEVAXlooks like and what the package contains

Powder for suspension

White lyophilized compact mass

The cardboard box contains four or ten ampoules and a patient leaflet.

Marketing authorization holder and manufacturer

BB-NCIPD Ltd., 1504 Sofia, 26, Y. Sakazov Blvd.

tel. +359 2 944 61 91

fax: +359 2 943 34 55

e-mail: bulbio@bulbio.com

Date of latest approval of the leaflet: March 2015

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CALGEVAX, прах за суспензия

( BCG за имунотерапия )

Моля, прочетете внимателно тази листовка, преди да започнете да приемате лекарството си, защото тя съдържа информация,

което е важно за вас.

- Запазете тази листовка; може да се наложи да я прочетете отново.

- Ако имате допълнителни въпроси, моля попитайте Вашия лекар или фармацевт.

- Това лекарство е предписано лично на Вас и не трябва да го давате на други хора. То

може да им навреди, дори ако техните симптоми са същите като Вашите.

- Ако забележите някакви нежелани реакции, моля, уведомете Вашия лекар или фармацевт. Това включва всички

възможни нежелани реакции, които не са посочени в тази листовка. Моля, вижте точка 4.

В тази листовка :

1.

Какво CALGEVAX е и за какво се използва за

2.

Какво трябва да знаете, преди да приемете CALGEVAX

3.

Как да приемате CALGEVAX

4.

Възможни нежелани реакции

5.

Съхраняване CALGEVAX

6.

Съдържание на опаковката и допълнителна информация

1.

Какво CALGEVAX е и за какво се използва за

CALGEVAX се BCG, специално проектиран за имунотерапия на рак.

• интравезикално:

- За профилактика на рецидивите на повърхностни тумори на пикочния мехур след трансуретрална

резекция;

- За лечение на карцином на място на пикочния мехур.

• перкутанно:

- За лечение на злокачествена меланома;

- За неспецифични адювантна имуностимулиращ терапия на други нововъзникнали злокачествени новообразувания

(На белия дроб, рак на гърдата, остра левкемия, лимфосарком или остеосарком).

2.

Какво трябва да се знае преди CALGEVAX се дава

Да не се прилага CALGEVAX ако:

-

има алергия към активното вещество или към някоя от останалите съставки на това лекарство (изброени в

т.6).

Предупреждения и предпазни мерки

Трябва да се консултирате с Вашия лекар или pharmacyst преди да използвате CALGEVAX.

CALGEVAX НЕ се прилага:

● при пациенти с имуносупресия;

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● в случай на пациенти с присъща или придобита имунна недостатъчност, причинена от заболяване или

антитуморна терапия;

● в случай на ХИВ-позитивни лица и пациенти на имуносупресия дози стероиди или други

имуноподтискащи;

● в фебрилни състояния;

● в случай на инфекция на пикочните пътища или масивна хематурия, както и в паралелна анти-микробно

терапия;

• В активна туберкулоза; положителен туберкулинов Манту тест е противопоказание само в случай на

сертифицирана активна туберкулозна инфекция;

• В случай на постоянно или предишната реакция системна BCG (т.е. системен грануломатозна болест,

която може да бъде разкрита, след като администрацията на BCG; тя се определя въз основа на

следните симптоми: хрема ≥39.5 ° С в продължение на ≥ 12 часа; треска ≥38.5 ° С в продължение на ≥48 часа, пневмония,

хепатит или друг орган дисфункция извън пикочо-половата система, включваща грануломатозен

възпаление, което е установено чрез биопсия, или класическите симптоми на сепсис).

Необходими предпазни мерки при употреба:

• Пациенти, които са имали лекарството се прилага интравезикално, трябва да поддържат адекватно

хидратация.

• интравезикалното приложение на CALGEVAX може да доведе до локално възпаление на пикочния

пикочния мехур, придружено с хематурия, дизурия, полакиурия и грипоподобни симптоми. Пациенти с

малък капацитет на пикочния мехур са изложени на висок риск от сериозни локални реакции.

• В случай на съмнение за системна BCG инфекция след прилагане на CALGEVAX, които

може да продължи с по-висока температура над 39 0 С, персистираща температура над 38 0 С или умора, бързо

лечение противотуберкулозните се извършва след консултация със специалист.

• CALGEVAX съдържа живи бактерии и трябва да бъдат третирани като инфекциозен материал. Лицата с

имуносупресия не трябва да се справят CALGEVAX.

• трябва да се извършва на катетеризация на пикочния мехур внимателно избягва traumatization на

лигавиците. Ако лекарят, извършване на лечение, реши, че катетеризация е

травматично (например, включва кървене), приложението на продукта трябва да бъде отложено

от най-малко 14 дни.

• В случай на сериозни общи нежелани реакции, интервалите между отделните администрации са

удължен. Лечението с CALGEVAX може да бъде прекратен по преценка на лекаря,

извършване на лечение. лечение противотуберкулозните се препоръчва в случай на дисеминирана

BCG инфекция.

Деца и юноши

CALGEVAX не се препоръчва за деца, поради липса на достатъчно данни за безопасността

и ефективността на продукта по отношение на деца.

Други лекарства и CALGEVAX:

Трябва да информирате Вашия лекар или pharmacyst ако сте приемали или е възможно да приемате други лекарства.

Прием на CALGEVAX може да доведе до повишена чувствителност към туберкулин, което би

усложни тълкуването на реакцията на кожата към туберкулин в бъдещо изследване по подозрения

микобактериални инфекции. В този контекст, се препоръчва да се определи реакция на пациента към

туберкулин преди имунотерапията с CALGEVAX.

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Бременност, кърмене и плодородие

Не приемайте това лекарство без предварителна консултация с Вашия лекар, ако сте бременна или кърмите,

мисля, че може да сте бременна или планирате да забременеете.

Балансът на съотношението полза / риск се оценява внимателно, когато лекарственият продукт трябва да се прилага

по време на бременност и кърмене, поради липса на клинични данни за неговото прилагане в тях

случаи. Известно е, че за кърмещи жени със системно BCG инфекция може да зарази детето.

Шофиране и работа с машини

Ефектът на CALGEVAX върху способността за шофиране и работа с машини не е проучена.

3.

Как да администрира CALGEVAX

Трябва да приемате това лекарство, както е описано в тази листовка или както Ви лекар на pharmacyst Ви е казал.

Ако не сте сигурни за нещо, попитайте Вашия лекар или pharmacyst.

Интравезикален ПРИЛАГАНЕ:

CALGEVAX се прилага най-малко 14 дни след извършване на биопсия, трансуретрална резекция

или травматична катетеризация. Прилагането на CALGEVAX трябва да се направи като се използва асептична техника

и под контрола на уролог, който има опит в прилагането на продукта. Три или

четири ампули на продукта, се използват за всеки интравезикална инстилация по преценка на уролога

извършване на лечение. Съдържанието на всяка ампула се разтваря в 1 мл стерилен физиологичен разтвор (а

разтвор на натриев хлорид с концентрация от 9 г / л), внимателно се разклаща до получаване на

хомогенна суспензия. Сместа се изтегля със спринцовка и след това се връща в ампулата три

пъти, за да се осигури пълно смесване, което свежда до минимум образуването на буци микобактериите. Най-

съдържанието на ампулите се прехвърля в спринцовка от 50 мл. Допълнителна обем на стерилен физиологичен разтвор

се добавя което общият обем 50 мл. Светлината намалява значително ефективността на

CALGEVAX. Следователно, излагане на пряка светлина трябва да се избягва преди и след

изготвяне на окачването. Суспензията трябва да се използва веднага.

След изпразване на пикочния мехур, на разтворения CALGEVAX се всели бавно в пикочния мехур през

уретрален катетър. Продуктът остава в пикочния мехур в продължение на два часа. Докато

CALGEVAX се в пикочния мехур, на всеки 15 минути пациента заместници следното

професии: полу-лявата струговане, полу-надясно по струговане, лявата страна, дясната страна.

Стандартната схема на приложение включва една интравезикално вливане веднъж седмично в течение

на шест седмици (индуциране на лечение). Поддържащата терапия се определя въз основа на всеки отделен случай. Най-

могат да бъдат използвани следните схеми: месечно в продължение на 6-12 месеца или минимални

три седмични вливания на 3-ти, 6-ти, 12-ти, 18-ти, 24-ти, 30-ти и 36-ти месец от датата на първото

вливане.

Перкутанно приложение:

Десет хоризонтална и десет вертикални линии са направени от иглата на спринцовката върху площ от 5/5 см. Най-

обезобразяване трябва да откъсне само на епидермалния слой, без да причинява значително кървене.

Устройство за multipuncture може да се използва за регулиране на дълбочината на обезобразяване.

място за обезобразяване се лекува чрез следния 0,5 мл суспензия: съдържанието на една ампула е

се смесва с 0,5 мл стерилен физиологичен разтвор за да се получи концентрация от 75 мг / мл. Една ампула се използва за един

обезобразяване. Преди процедурата, площта трябва да бъдат лекувани с ацетон, след което неговото пълно

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изпаряване трябва да се очаква. А нова област е избрана за всяка следваща обезобразяване. честотата

и продължителност на приложение се определя от лекаря, извършване на лечение.

4. Възможни нежелани реакции:

Това лекарство, както и повечето други лекарства, може да причини нежелани реакции, въпреки че това не се отнася за

всички хора.

Интравезикално администрация

• Локални реакции: преходен дизурия, полакиурия, хематурия, бактериална инфекция на пикочния

тракт, грануломатозен простатит, епидидимит, орхит.

• Общи реакции: отпадналост, повишена температура, треска, изпотяване, гадене, повръщане,

главоболие, мускулни болки, колики, дифузен обрив, чернодробна токсичност, дисеминирана BCG

инфекция, генерализирана свръхчувствителност, еритема nodosium, конюнктивит, увеит, витилиго,

артрит, левкопения или панцитопения, спленомегалия.

В случай на мехура администрация, нежелани лекарствени реакции (затруднено уриниране, полакиурия, треска) се появяват 3-4 часа

след вливане и са преходни - те продължават по 24-72 часа. А локализиран (epididymistis, орхит,

простатит) или системна BCG инфекция се появява по-рядко.

перкутанно приложение

• Локални реакции: локализиран сърбеж или обрив, болезнени язви на мястото на приложение,

регионална аденопатия.

• Общи реакции: умора, повишена температура, треска, изпотяване, гадене, повръщане,

главоболие, мускулни болки, колики, дифузен обрив, чернодробна токсичност, дисеминирана BCG

инфекция, генерализирана свръхчувствителност, еритема nodosium, конюнктивит, увеит, витилиго,

артрит, левкопения или панцитопения, спленомегалия.

В случай на сериозни нежелани реакции, интервалите между отделните администрации са разширени.

Лечението с CALGEVAX може да бъде прекратен по преценка на лекаря, извършващ

лечение. лечение противотуберкулозните се препоръчва в случай на дисеминирана BCG инфекция.

Пациентът трябва да информират своя лекар за всякакви странични ефекти, не посочи в листовката.

Докладване нежелани реакции

Ако забележите някакви нежелани реакции, моля, уведомете Вашия лекар или фармацевт. Това включва всички възможни

нежелани лекарствени реакции, които не са посочени в тази листовка. Можете също така да се съобщава за нежелани реакции

директно чрез Националната система за докладване, посочи по-долу:

Българска агенция по лекарствата

8, Дамян Груев St., 1303 София,

тел .: + 359 28903417, уебсайт: www.bda.bg

Чрез отчетна нежелани реакции може да допринесе за осигуряване на повече информация относно безопасността на

това лекарство.

5. Съхранение на CALGEVAX

Да се ​​съхранява при температура от 2 ° С до 8 ° С.

Да се ​​съхранява в картонената опаковка, за да се избегне излагането на светлина.

Да се ​​пази далеч от деца.

След ресуспендиране, се избягва излагането на светлина и администриране незабавно.

---

Page 5

5

Продуктът не трябва да се използва след изтичане на срока им, посочи върху опаковката.

Продуктът е годен за употреба до последния ден на месеца, посочен.

Специални мерки при изхвърляне

Неизползваните количества от окачването, игли, спринцовки и катетри са унищожени в съответствие с

правилата за изхвърляне на заразни материали. В случай на разлив, районът се почиства с 70% разтвор на

етилов алкохол.

6. Съдържание на опаковката и допълнителна информация

Съдържание на CALGEVAX

Активно вещество - Mycobacterium говежди BCG (Bacillus Calmette-Guérin) 11,25 мг (37,5 мг полу-

суха бактериална маса) 1,0 -3,0 x10 8 жизнеспособни единици

Помощни вещества: Натриев глутамат 40 мг

Какво CALGEVAXlooks харесва и какво пакетът съдържа

Прах за окачване

White лиофилизирана компактна маса

картонената кутия съдържа четири или десет ампули и листовка за пациента.

Притежател на разрешението и производител

BB-НЦЗПБ ЕООД, София 1504, 26, Я. Сакъзов бул

тел. +359 2 944 61 91

факс: +359 2 943 34 55

Е-мейл : bulbio@bulbio.com

Дата на последно одобрение на листовката: Март 2015

========================================================
https://www.ncbi.nlm.nih.gov/labs/articles/2807782/

ПАТЕНТ КАТРАПС - http://www.google.tl/patents/WO2006122380A1?cl=en

Antitumor agent on the base of bcg vaccine, method for his preparation and its use 
WO 2006122380 A1

РЕЗЮМЕ

The invention relates to antitumor agent on the base of BCG vaccine, the method his preparation and its application in patients diagnosed with oncological diseases. The agent according the invention is a mixture of two individual fractions: Fraction 1 - Live BCG bacteria and Fraction 2 - Extract-Filtrate of BCG Vaccine where the ratio between them may vary from 1:0.01 to 0.01:1. The agent is used in the treatment of all forms of cancer at all stages of the progress of the disease. The agent according the invention allows maximum opportunity for action of the BCG vaccine antitumor agent to terminate of fast and unchecked proliferation of tumor cells, reduced toxicity and blocking of the inhibitive effect of tumor cells on normal cells in the body.

ПРЕТЕНЦИИ  (текстът с оптично разпознаване на знаци може да съдържа грешки)

PATENT CLAIMS

1. An antitumor agent developed on the basis of BCG vaccine, containing standard solution of BCG vaccine with Mycobacterium bovis as active substance, that is characterized with the fact, that is composed of two different fractions - Fraction 1 "Live BCG Bacteria", containing a standard BCG vaccine diluted once by physiological solution or distilled water to a concentration from 100 to 10 μg BCG and secondly by physiological solution to a concentration from 10 to 0.001 μg/ml and Fraction 2 "BCG Vaccine Extract-filtrate", containing standard solution of BCG vaccine, diluted by 4-6 ml distilled water or 3-5 ml physiological solution, at ratio Fraction 1 : Fraction 2 from 1 :0.01 to 0.001 : 1.

2. A method for preparation a substance with antitumor activity based on standard solution of BCG vaccine with Mycobacterium bovis as active agent that is characterized with the fact, that are given two different fractions, termed as Fraction 1 "Live BCG Bacteria", obtained through double dilution of one vial standard BCG vaccine by physiological solution or distilled water and Fraction 2 "BCG Vaccine Extract- filtrate", obtained by adding a physiological solution or distilled water to a standard solution of BCG vaccine and subsequent treatment of the obtained suspension with ultrasound prior to refrigerate it from 2 to 60 days, preferably from 2 to 20 days, whilst controlling the live bacteria contents in the two fractions, which at the end are mixed at a ratio of Fraction 1 : Fraction 2 from 0.01 : 1 to 1:0.01 .

3. A method, according to claim 2, that is characterized with the fact, that Fraction 1 is obtained as standard solution of BCG vaccine, which is once diluted by physiological solution or distilled water to a concentration from 100 to 10 μg/ml and secondly by physiological solution from 10 to 0.001 μg/ml.

4. A method, according to claim 2, that is characterized with the fact, that Fraction 2 obtaines after to a standard solution of BCG vaccine are added 3 to 5 ml physiological solution or 4 to 6 ml distilled water and the resulting suspension, after ultrasound treatment for 8 to 12 minutes, refrigerates from 7 to 60 days, preferably from 2 to 20 days, at a temperature of 4-8⁰C, whilst periodically have controlled for presence of live BCG bacteria, after that the resulting suspension filters through a sterile filter and preserves in a refrigerator at 4-8⁰C.

5. Use of the BCG-vaccine based agent with antitumor activity, which applies intradermally to oncological patients in quantities between 0.1 and 0.2ml for treatment of a solid tumors and metastases originating from them in all systems of the human body, as well as for the prevention of relapses after surgical treatment, radiological treatment and chemotherapy.

6. Use, according to claim 5, for treatment of various forms of cancer and specifically cancer of the throat, breast cancer, stomach cancer, liver cancer, cancer of the uterus, cancer of the colon, skin cancer and any other type of solid tumors, brain tumors, malignant melanoma, as well as of the metastases originating from them in all systems of the human body.

ОПИСАНИЕ  (текстът с оптично разпознаване на знаци може да съдържа грешки)

ANTITUMOR AGINT BASED ON BCG VACCINE, METHOD OF ITS PREPARATION AND USE

Technical area

The invention relates to an agent with antitumor activity on the basis of BCG vaccine, a method for its preparation and its application for treatment of patients diagnosed with oncological diseases.

Background of the invention

The results of a number of scientific studies published over the past several years show that the antitumor properties of the Mycobacteriaceae family are due to active agents it contains or produces.

Thus, US patent 6, 274, 356 describes a carbohydrate complex, in essence representing a complex of polysaccharides, extracted from Mycobacterium vaccae, which shows high antitumor activity in the treatment of different oncological diseases. The method of preparation of the carbohydrate complex has also been described as a series of actions, involving obtaining precipitates and dialisates by extraction and purification.

Summary of the invention

The problem of the present invention is obtaining of an agent with antitumor activity based on the BCG vaccine, which gives maximum opportunity for the action of the antitumor factor of the BCG vaccine in order to terminate the rapid and uncontrolled proliferation of tumor cells in an environment of reduced toxicity and blocking of the inhibitive effect of tumor cells on normal cells in the body as well the method of obtaining of BCG-based agent with antitumor activity and its use. According to the invention this problem was solved with a BCG vaccine based agent (BCG with Mycobacterium bovis as active substance, controlled by the manufacturer in compliance with WHO requirements regarding the physical, chemical and biological properties of the bacterial strains and of the final product), which has a powerful inhibitive effect on tumor cells and has demonstrated no undesirable side effects.

The agent according to the invention is a mixture of two independent fractions the ratio between which may vary from 1 :0.01 to 0.01:1.

Fraction 1 named "Live BCG Bacteria" contains a standard BCG vaccine, diluted once by physiological solution or distilled water to a concentration from 100 to 10 μg/ml and secondly diluted by physiological solution from 10 to 0.001 μg/ml.

Our definition of standard BCG vaccine solution means 1 ampoule BCG vaccine containing 500 μg dry substance, diluted by 1 ml distilled water used as standard solvent.

Fraction 2 termed "BCG Vaccine Extract-filtrate" contains standard solution of BCG vaccine, diluted by 4-6 ml distilled water or 3-5 ml physiological solution.

Another object of the invention is a method for preparation of an agent with antitumor activity as per the invention. The essence of the method lies in obtaining two different fractions, respectively termed Fraction 1 - "Live BCG Bacteria" and Fraction 2 - "BCG Vaccine Extract-filtrate", which under specific processing conditions are mixed in a certain proportion.

Fraction 1 is made from a standard ampoule of 500 μg dry substance of BCG vaccine, the shelf-life (exp.date) of which must not have expired. Following the instructions the ampoule is diluted with standard solvent whilst strictly observing sterility rules in a sterile laboratory box, treated by UV lamps for 18-24 hours in advance. The resulting suspension is additionally diluted with physiological solution or distilled water up to a concentration of 100 to 10 μg/ml BCG. This suspension is diluted again until a concentration of 10 to 0.001 μg/ml is obtained (accepted as 10 provisional units according in line with the invention).

The content of live BCG bacteria is controlled by periodic microbiological samples on Leuvenstein-Jensen media for tuberculosis bacteria containing (in g/1 to double distilled water):

L-Aspargine 3.6

Monocalcium phosphate 2.4

Magnesium sulfate 0.24

Magnesium citrate 0.6

Potato flour 30.0

Malachite green 0.4

Glycerol 12.0 ml

Eggs 1 litre pH 7.2 ± 0.2 at 25 ⁰C

Growth is controlled by the presence of specific colonies and specific morphology of the bacteria stained by Ziehl-Nielsen and examined under the microscope.

Fraction 2 is generated from standard BCG vaccine by adding 3 to 5ml physiological solution or 4 to 6 ml distilled water and ultrasound treatment for 8 to 12 minutes. The resulting suspension is left for a period of 2 to 60 days, preferably 2 to 20 days in a refrigerator at a temperature of 4-8⁰C.

The content of live BCG bacteria is controlled by periodic microbiological samples on Leuvenstein-Jensen media for tuberculosis bacteria and the growth is demonstrated by the presence of specific colonies and specific morphology of the bacteria stained by Ziehl-Nielsen and examined under the microscope.

Between the 2^nd and the 60^th day the suspension is filtered though a sterile filter and the filtrate obtained is stored in a refrigerator at a temperature of 4 to 8⁰C as initial extract-filtrate, which contains no live BCG bacteria.

Fraction 2 is a clear solution without any visible impurities, colour or odour. The chemical components of Fraction 2 include proteins, nucleoproteides, free aminoacids, polysaccharides, and small quantity of fats with the ratio between individual ingredients varying uncontrollably.

After the control examination aimed to establish the presence of live bacteria, the two fractions are mixed at Fraction 1 to Fraction 2 ratio from 0.01:1 to 1:0.01.

Yet another aspect of the invention is the use of the BCG-vaccine based agent with antitumor properties. The agent as per the invention is used for the treatment of all forms of malignancies in all stages of the disease.

The antitumor acting substance as per the invention is further applied for the treatment of various forms of cancer and specifically cancer of the throat, breast cancer, stomach cancer, liver cancer, cancer of the uterus, cancer of the colon, brain tumors, malignant melanoma, skin cancer and any other type of solid tumors, as well as of the metastases originating from various solid tumors in all systems of the human body.

Immediately after mixing the two fractions in the ratio determined as per the invention the BCG vaccine based agent with antitumor activity is injected intradermally to patients with cancer in a ratio from 0.1 to 0.2 ml.

For the purpose of sustained use the agent as per the invention is transferred to an ice bath and inserted into vacuum ampoules of 0.2 ml. Advantages of the Invention

As per the invention the main advantages of the invention of the areHT with antitumor effect are:

(1) powerful effect on tumor cells and termination of their rapid and uncontrolled proliferation;

(2) powerful effect on metastases, stopping their growth and leading to their regression and disappearance;

(3) blocking the repressing effect of tumor cells on normal cells in the system;

(4) low toxicity;

(5) possibly acts as cell growth factors, controlling the non- differentiated proliferation of cancer cells;

(6) simplified method of obtaining the agent.

Example for implementation of the invention The invention can be illustrated by the examples below, which clarify it, without prejudice to the scope of its protection.

Example 1

Obtaining Fraction 1 - "Live BCG Bacteria"

One standard BCG vaccine ampoule (manufactured by BUL BIO National Center of Infectious and Parasitic Diseases, Sofia) within the designated shelf-life, containing 500 μg dry substance is opened in sterile laboratory box treated with UV lamps for 18 hours in advance. One ml of distilled water is added and the resulting suspension is diluted with physiological solution to a concentration of 100 μg BCG in 1 ml physiological solution. The resulting suspension is further diluted to 10 μg/ml BCG. The content of live BCG bacteria is controlled by periodic microbiological samples on Lowenstein-Jensen media for tuberculosis bacteria. The growth is demonstrated by the presence of specific colonies 6 000011

and specific morphology of the bacteria stained by Ziehl-Melsen and examined under the microscope.

Obtaining Fraction 2 - "BCG Vaccine Extract-filtrate"

One standard BCG vaccine vial is opened in a sterile laboratory box treated with UV lamps in advance. One ml of a standard vaccine solvent is added. Three ml physiological solution are added to the resulting suspension, which is then treated by ultrasound for 8 min and refrigerated at a temperature of 4 to 8 ⁰C for 2 days. On the 1^st and the 2^nd day the suspension is periodically controlled by microbiological samples on Lowenstein- Jensen media and then filtered though sterile filter Falcon® where the pores are < 0.1 mm and the filtrate obtained is a ready-for-use Fraction 2.

Final preparation of the Antitumor Agent

The two fractions obtained in the manner described above are mixed under sterile conditions in a sterile container in the ratio = Fraction 1 : Fraction 2 as 1 : 0.01 where the resulting mixture is a ready-for-use agent as per the invention.

The agent with antitumor activity as per the invention is applied to the patients strictly intradermally on the volar part of the hand as Mantoux in a quantity of 0.1 ml.

Example 2

Obtaining Fraction 1 - "Live BCG Bacteria"

One standard BCG vaccine ampoule within the designated shelf- life, containing 500 μg dry substance is opened in sterile laboratory box treated with UV lamps for 20 hours in advance and ImI of distilled water is added. The resulting suspension is diluted with distilled water to a BG2006/000011

concentration of 50 μg/ml. The resulting suspension is additionally diluted with physiological solution to a concentration of 0.1 μg/ml.

The presence of live BCG bacteria is controlled through periodic microbiological samples as described in Example 1.

Fraction 2 - "BCG Vaccine Extract-filtrate"

One standard BCG vaccine vial is opened in a sterile laboratory box treated with UV lamps in advance. Then 1 ml of standard vaccine diluent is added. To the resulting suspension 4 ml physiological solution is added. The resulting suspension is then treated by ultrasound for 9 min and refrigerated at a temperature of 4 to 8⁰C for 10 days whilst periodically controlled for the presence of live bacteria as described in Example 1. On the 10^th day the suspension is filtered though a sterile filter Falcon® where the pores are < 0.1 mm and the filtrate obtained is a ready-for-use Fraction2.

Final preparation of the Antitumor Agent

The two fractions obtained in the manner described above are mixed under sterile conditions in a sterile container in the ratio = Fraction 1 : Fraction 2 equal to 1 : 0.1 where the resulting mixture is a ready-for-use agent as per the invention.

The agent with antitumor activity as per the invention is applied strictly intradermally on the volar part of the hand as Mantoux in a quantity of 0.1 ml.

Example 3

Obtaining Fraction 1 - "Live BCG Bacteria"

One standard BCG vaccine ampoule is diluted with standard vaccine solvent in a sterile laboratory box, treated by UV lamp for 22 hours in advance, as described in Example 1. The obtained suspension is diluted with physiological solution to a concentration of 10 μg/ml. The resulting suspension is additionally diluted with physiological solution to a concentration of 0.001 μg/ml.

The presence of live BCG bacteria is controlled as described in Example 1 above.

Fraction 2 - "BCG Vaccine Extract-filtrate"

One standard BCG vaccine vial is opened in a sterile laboratory box treated with UY lamps in advance and after that 1 ml distilled water is added. The obtained suspension is then diluted by 5 ml physiological solution, which is treated by ultrasound for 10 min and refrigerated at a temperature of 4 to 8⁰C for 20 days whilst periodically controlled for the presence of live bacteria as described in Example 1. On the 20^th day the suspension is filtered though sterile filter Falcon® where the pores are <0.1 mm. The resulting filtrate doesn't contain live BCG bacteria and is a ready-for-use Fraction 2.

Final preparation of the Antitumor Agent

The two fractions obtained in the manner described above are mixed under sterile conditions in a sterile container in the ratio = Fraction 1 : Fraction 2 equal to 1 : 1. During transportation the suspension is transferred to an ice bath and then into vacuum ampoules of 0.2 ml and refrigerated at 4-8⁰C.

The resulting agent with antitumor activity as per the invention is fit for use within 1 month of production.

The agent with antitumor activity as per the invention is applied to the patients with oncological disease strictly intradermally on the volar part of the hand as Mantoux in a quantity of 0. ImI. Example 4

Obtaining Fraction 1 - "Live BCG Bacteria"

In a sterile laboratory box treated by UV lamp for 24 hours in advance one standard ampoule of BCG vaccine is diluted with a standard vaccine solvent as described in Example 1. The obtained suspension is diluted with physiological solution to a concentration of 10 μg/ml. The resulting suspension is additionally diluted with physiological solution to a concentration of 0.001 μg/ml.

The presence of live BCG bacteria is controlled as described in Example 1 above.

Obtaining Fraction 2 - "BCG Vaccine Extract-filtrate"

One standard BCG vaccine vial is opened in a sterile laboratory box treated with UV lamps in advance and after that 1 ml distilled water is added. Then the obtained suspension is diluted with 6 ml distilled water which is treated by ultrasound for 12 min and refrigerated at a temperature of 4 to 8⁰C for 40 days whilst periodically controlled for the presence of live bacteria as described in Example 1. On the 40^th day the suspension is filtered though a sterile filter Falcon® where the pores are < 0.1 mm. The resulting filtrate doesn't contain live BCG bacteria and is a ready-for-use Fraction 2.

Final preparation of the Antitumor Agent

The two fractions obtained in the manner described above are mixed under sterile conditions in a sterile container in the ratio = Fraction 1 : Fraction 2 equal to 0.1 : 1.

The agent with antitumor activity as per the invention is applied strictly intradermally on the volar part of the hand as Mantoux in a quantity of 0.1 ml. 6 000011

10

Example 5

Obtaining Fraction 1 - "Live BCG Bacteria"

In a sterile laboratory box treated by UV lamp for 23 hours in advance one standard ampoule of BCG vaccine is diluted with standard vaccine solvent as described in Example 1. The obtained suspension is diluted with distilled water to a concentration of 20 μg/ml. The resulting suspension is additionally diluted with physiological solution to a concentration of 0.01 μg/ml.

The presence of live BCG bacteria is controlled as described in Example 1 above.

Obtaining Fraction 2 - "BCG Vaccine Extract-filtrate"

One standard BCG vaccine vial is opened in a sterile laboratory box treated with UV lamps in advance and 1 ml distilled water is added. Then, the obtained suspension is diluted with 4 ml physiological solution, which is treated by ultrasound for 10 min and refrigerated at a temperature of 4 to 8⁰C for 60 days whilst periodically controlled for the presence of live bacteria as described in Example 1. On the 60^th day the suspension is filtered though a sterile filter Falcon® where the pores are < 0.1 mm. The resulting filtrate doesn't contain live BCG bacteria and is a ready-for-use Fraction 2.

Final preparation of the Antitumor Agent

The two fractions obtained in the manner described above are mixed in a sterile container under sterile conditions in the ratio = Fraction 1 : Fraction 2 equal to 0.01 : 1.

The agent with antitumor activity as per the invention is applied strictly intradermally on the volar part of the hand as Mantoux in a quantity of 0.2 ml. _u T/BG2006/000011

Examples of Clinical Tests of the Agent with Antitumor Activity as per the Invention

In Brain Tumors

Example 6

Patient B. V., 57 year-old male diagnosed with Craniopharingeoma. Partial resection performed. At that time the patient complained of severe headache, disturbances of the conscious, visual impairments and motor weakness when moving. The ensuing treatment was terminated due to deterioration of symptoms. Soon after the first administration of the agent with antitumor activity as per the invention visible improvement in the patient's condition could be observed to the gradual complete disappearance of symptoms. The patient is now leading a normal, healthy life.

Example 7

Patient, M. G., 24 year-old male diagnosed with Meduloblastoma causing spastic paralysis of the right leg and full patient immobilization. Within 6 months after the first administration of the agent as per the invention the paralysis gradually disappears and full recovery is gradually achieved.

Example 8

Patient S. T. 21 year-old female diagnosed with Ewing's Sarcoma. The therapy with the agent as per the invention commenced after surgical treatment and chemotherapy. At this stage crawling paralysis of the patient's leg is present and the patient was immobilized in a wheelchair. During the course of treatment with the agent as per the invention she regained the ability to walk on her own without assistance. In Cancer of the Urinary Bladder

Example 9

Patient D.K., 51 year-old diagnosed with bladder cancer is subjected to Trans-Urethral Resection (TUR) every 3-4 months and subsequently went through surgery due to massive chematuria. Several months after administration of the agent as per the invention the patient is in good condition and leads a normal life.

Example 10

Patient A. A., 46 year-old male, diagnosed with bladder cancer in bad general condition due to profuse chematuria. Soon after administration of the agent as per the invention the patient's condition improved and no chematuria is presently observed. Several subsequent cytoscopies showed negative results for the presence of tumor cells.

Example 11

Patient CC, 57 year-old male, physician, diagnosed with bladder cancer. TUR is applied once in 2-3 months. After administration of the agent as per the invention the chematuria disappeared and the following cytoscopies showed negative results for the presence of tumor cells.

In Skin Cancer

Example 12

Patient N.T., diagnosed with melanoma malignum. After application of the agent the melanoma spot disappeared. BG2006/000011

13

Example 13

Patient G.G., diagnosed with Angiosarcoma with large tumor mass in the area of the armpit, which gradually disappeared after administration of the agent as per the invention leaving only a crust.

Example 14

Patient D.D. diagnosed with Sarcoma and large tumor mass in the area of the arm. Following treatment with the agent as per the invention for several months the tumor disappeared leaving only a scar.

In Lung Cancer

Example 15

Patient LT. , male, diagnosed with Ca pulmonum  - http://www.google.tl/patents/WO2006122380A1?cl=en

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