WO1993013403A1 - FLUORESCENCE DIAGNOSTICS OF CANCER USING δ-AMINO LEVULINIC ACID - Google Patents
FLUORESCENCE DIAGNOSTICS OF CANCER USING δ-AMINO LEVULINIC ACID Download PDFInfo
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- WO1993013403A1 WO1993013403A1 PCT/SE1992/000879 SE9200879W WO9313403A1 WO 1993013403 A1 WO1993013403 A1 WO 1993013403A1 SE 9200879 W SE9200879 W SE 9200879W WO 9313403 A1 WO9313403 A1 WO 9313403A1
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- fluorescence
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- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 41
- 201000011510 cancer Diseases 0.000 title claims abstract description 14
- ZGXJTSGNIOSYLO-UHFFFAOYSA-N 88755TAZ87 Chemical compound NCC(=O)CCC(O)=O ZGXJTSGNIOSYLO-UHFFFAOYSA-N 0.000 title claims abstract description 6
- 238000000034 method Methods 0.000 claims abstract description 20
- 238000001514 detection method Methods 0.000 claims abstract description 12
- 239000002674 ointment Substances 0.000 claims abstract description 12
- 230000005284 excitation Effects 0.000 claims abstract description 10
- 239000000835 fiber Substances 0.000 claims description 7
- 239000000126 substance Substances 0.000 claims description 6
- 238000011835 investigation Methods 0.000 claims description 5
- 238000003384 imaging method Methods 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 230000003595 spectral effect Effects 0.000 claims description 3
- ZLHFONARZHCSET-UHFFFAOYSA-N 5-aminolevulinic acid hydrochloride Chemical group Cl.NCC(=O)CCC(O)=O ZLHFONARZHCSET-UHFFFAOYSA-N 0.000 claims description 2
- 230000037396 body weight Effects 0.000 claims description 2
- 238000012544 monitoring process Methods 0.000 claims description 2
- 230000003211 malignant effect Effects 0.000 claims 1
- 239000003795 chemical substances by application Substances 0.000 abstract description 9
- 238000010253 intravenous injection Methods 0.000 abstract description 4
- 230000036039 immunity Effects 0.000 abstract 1
- SSOLNOMRVKKSON-UHFFFAOYSA-N proguanil Chemical compound CC(C)\N=C(/N)N=C(N)NC1=CC=C(Cl)C=C1 SSOLNOMRVKKSON-UHFFFAOYSA-N 0.000 abstract 1
- 239000000439 tumor marker Substances 0.000 abstract 1
- 210000001519 tissue Anatomy 0.000 description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 230000005855 radiation Effects 0.000 description 4
- 206010004146 Basal cell carcinoma Diseases 0.000 description 3
- 241000700159 Rattus Species 0.000 description 3
- 210000003205 muscle Anatomy 0.000 description 3
- 206010006187 Breast cancer Diseases 0.000 description 2
- 208000026310 Breast neoplasm Diseases 0.000 description 2
- KSFOVUSSGSKXFI-GAQDCDSVSA-N CC1=C/2NC(\C=C3/N=C(/C=C4\N\C(=C/C5=N/C(=C\2)/C(C=C)=C5C)C(C=C)=C4C)C(C)=C3CCC(O)=O)=C1CCC(O)=O Chemical compound CC1=C/2NC(\C=C3/N=C(/C=C4\N\C(=C/C5=N/C(=C\2)/C(C=C)=C5C)C(C=C)=C4C)C(C)=C3CCC(O)=O)=C1CCC(O)=O KSFOVUSSGSKXFI-GAQDCDSVSA-N 0.000 description 2
- 206010027476 Metastases Diseases 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 238000001917 fluorescence detection Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 230000037311 normal skin Effects 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 150000004032 porphyrins Chemical class 0.000 description 2
- 229950003776 protoporphyrin Drugs 0.000 description 2
- 239000010453 quartz Substances 0.000 description 2
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 208000005440 Basal Cell Neoplasms Diseases 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 206010055113 Breast cancer metastatic Diseases 0.000 description 1
- MYMOFIZGZYHOMD-UHFFFAOYSA-N Dioxygen Chemical compound O=O MYMOFIZGZYHOMD-UHFFFAOYSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 208000009956 adenocarcinoma Diseases 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000005907 cancer growth Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 235000017168 chlorine Nutrition 0.000 description 1
- 125000001309 chloro group Chemical class Cl* 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 238000002405 diagnostic procedure Methods 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000011846 endoscopic investigation Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000001061 forehead Anatomy 0.000 description 1
- 150000003278 haem Chemical class 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 210000003470 mitochondria Anatomy 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 231100001078 no known side-effect Toxicity 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 229940109328 photofrin Drugs 0.000 description 1
- 239000003504 photosensitizing agent Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 238000005086 pumping Methods 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000007704 transition Effects 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/0019—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules
- A61K49/0021—Fluorescence in vivo characterised by the fluorescent group, e.g. oligomeric, polymeric or dendritic molecules the fluorescent group being a small organic molecule
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0071—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by measuring fluorescence emission
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0075—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence by spectroscopy, i.e. measuring spectra, e.g. Raman spectroscopy, infrared absorption spectroscopy
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0082—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes
- A61B5/0084—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes for introduction into the body, e.g. by catheters
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/0059—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence
- A61B5/0082—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes
- A61B5/0091—Measuring for diagnostic purposes; Identification of persons using light, e.g. diagnosis by transillumination, diascopy, fluorescence adapted for particular medical purposes for mammography
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B5/00—Measuring for diagnostic purposes; Identification of persons
- A61B5/43—Detecting, measuring or recording for evaluating the reproductive systems
- A61B5/4306—Detecting, measuring or recording for evaluating the reproductive systems for evaluating the female reproductive systems, e.g. gynaecological evaluations
- A61B5/4312—Breast evaluation or disorder diagnosis
Definitions
- Cancer diseases are very widespread and have a major social and socio-economic impact. The incidence of cancer has shown a steady increase during the last tens of years. It is generally accepted that an early cancer diagnosis and a radical treatment is of great importance for the prognosis for the patient.
- Conventional diagnostic methods for cancer comprise different types of X-ray investigations, endoscopic investigations with biopsy sampling in the hollow organs of the body, cytological puncture of resistances, as well as smear cytology.
- the present invention describes the use of ⁇ -amino levulinic acid (abbreviated ALA below) as a specifically tumour seeking agent, to be visualized through its specific fluorescence on irradiation with a laser or other suitable radiation source.
- ALA ⁇ -amino levulinic acid
- a point monitoring or an imaging fluorescence system can be used for the detection.
- ALA intravenously administered, or applied as an unguent over superficial tumours, is very selectively taken up by tumour cells, in which the substance is transformed into protoporphyrin IX, which exhibits a very strong and characteristic fluorescence in the red wavelength region.
- This procedure can be used, e.g. for the detection of early bronchial or bladder cancer and for the demarcation of the extension of a tumour for assuring a radical tumour treatment.
- tumour marking can be used in two ways, for diagnostics thorugh the fluorescence of the agent and for tumour treatment by photodynamic techniques, where the photo- excited agent transfers its energy to oxygen molecules, that make a transition from their normal triplet state to an aggressive singlet state. The release of singlet oxygen selectively in the tumour causes tissue necrosis.
- Photofrin Quadra Logic Technologies, Vancouver, Canada
- the present invention is based on our observation, in animal experiments as well as clinically, that the agent ⁇ -amino levulinic acid is an extremely efficient fluorescence marker for tumours. This is true for intravenous injection as well as for superficial treatment with ALA unguent direct on the tumour.
- the small non-fluorescing ALA molecules penetrate the cell membrane of malignant tumour cells and undergo in the mitochondria parts of the haem cycle, leading to the formation of protoporphyrin IX. Fluorescence can be induced by illumination by radiation from, e.g. a pulsed laser. This can be a dye laser pumped by a nitrogen laser. An especially suitable excitation wavelength is 405 nm, where porphyrins have their absorption maximum.
- the radiation can be conducted to the tissue through a quartz fibre, which can also be used to collect fluorescence light and to conduct it to a spectral analyzer.
- a quartz fibre which can also be used to collect fluorescence light and to conduct it to a spectral analyzer.
- This can be a spectrometer equipped with a diode-array detector or a more simple instrument based on optical filters.
- Tumours are marked by the occurrence of a fluorescence peak at 635 nm and a weaker peak at 710 nm.
- a demarcation ratio (the ratio between the specific fluorescence signal in a tumour and the signal in close-lying normal tissue) of about 10 or better can be obtained using this naturally occurring molecule in the body. It has no known side effects.
- a strong tumour demarcation is obtained after intra-venous injection if the investigation is performed about 1/2 hour after injection.
- ALA can also be prepared in an unguent base (about 20 % ALA) and be spread over superficial tumours
- An especially effective form of diagnostics for ultraviolet or violet excitation is to evaluate the background-free fluorescence at 635 nm and divide it by the fluorescence intensity at about 470 nm. Such a ratio is dimensionless and therefore the signal will be independent of the measurement distance, the angle of incidence for the light, and fluctuations in the excitation and detection equipment.
- the procedure given in this invention can also be used in combination with an imaging cancer detection system, which has been described in the Swedish patent # 455646.
- a simultaneous recording of images in three different fluorescence colours is then made (635 nm, 600 nm and 470 nm).
- An image intensifier which is activated in synchronism with the exciting light source, is used for the recording.
- a new, generalized and tumour enhancing image is obtained by subtracting the 600 nm image (background) from the 635 nm image, followed by a division of the result by the blue image, pixel by pixel.
- Cancer detection with fluorescence techiques using ALA is illustrated in Figure 1, where 1 indicates the excitation source, 2 indicates optical components to bring the exciting light to the tissue, 3 indicates the tissue that has been exposed to ALA, 4 indicates optical components for conducting the fluorescence light to a wavelength-dividing system 5 placed in front of the detection system 6. Finally, 7 indicates a signal processing system for tumour recognition based on the fluorescence properties.
- the system proposed enables strongly improved possibilities for early detection of malignant tumours, also small tumours in the hollow organs of the body, and for clear demarcation of diseased tissue from surrounding normal tissue. No side effects occur.
- a patient with basal cell carcinoma of the forehead was investigated with the method described in this invention. 6 hours before the fluorescence investigation the tumour areas were exposed to an unguent containing ALA in hydrochloric form (Porphyrin Products, Inc., Logan, Utah; catalogue # A 167). The unguent also covered a 10 mm wide zone of normal tissue.
- Fluorescence light was collected through the same fibre, and after passing a dichroic beam splitter, used for injecting the laser light into the fibre, the fluorescence light was focussed on the entrance slit of a Jobin-Yvon 0.25 m grating monochromator equipped with an EG&G Optical Multichannel Analyser (OMA) HI system.
- Huorescence spectra from a basal cell cancer tumour and from close-lying, ALA-exposed normal skin are shown in Figure 2.
- Using the intensity of the characteristic red fluorescence a demarcation ratio of 50 between the tumour and the normal tissue is obtained. If instead the ratio between the red and the blue- green fluorescence is used a demarcation ratio of 150 is obtained.
- Figure 1 Schematic arrangment for fluorescence detection with ALA.
- Figure 2 Fluorescence recordings for basal cell carcinoma and normal tissue for a patient that was exposed to an ALA unguent in the tumour area.
- Figure 3 Fluorescence recordings for metastasis of breast cancer and normal tissue for a patient that was exposed to an ALA unguent in the tumour area.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Epidemiology (AREA)
- Medicinal Chemistry (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Pathology (AREA)
- Heart & Thoracic Surgery (AREA)
- Medical Informatics (AREA)
- Molecular Biology (AREA)
- Surgery (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
A procedure for the detection of malignant tumours using the agent δ-amino levulinic acid (hydrochloride) as a tumour marker is described. The tissue is exposed to the agent either by intravenous injection or by application of an unguent over superficial tumours. After a suitable time delay fluorescence is induced in the tissue using an excitation source, e.g. a laser. Cancer then emerges with a strong and specific fluorescence peak at 635 nm. By dividing this red peak intensity by the intensity of the blue-green autofluorescence from the tissue an improved demarcation is obtained and also immunity to variations in many experimental parameters.
Description
Fluorescence diagnostics of cancer using δ-amino levulinic acid
Field of the invention
Cancer diseases are very widespread and have a major social and socio-economic impact. The incidence of cancer has shown a steady increase during the last tens of years. It is generally accepted that an early cancer diagnosis and a radical treatment is of great importance for the prognosis for the patient. Conventional diagnostic methods for cancer comprise different types of X-ray investigations, endoscopic investigations with biopsy sampling in the hollow organs of the body, cytological puncture of resistances, as well as smear cytology. The present invention describes the use of δ-amino levulinic acid (abbreviated ALA below) as a specifically tumour seeking agent, to be visualized through its specific fluorescence on irradiation with a laser or other suitable radiation source. A point monitoring or an imaging fluorescence system can be used for the detection. ALA, intravenously administered, or applied as an unguent over superficial tumours, is very selectively taken up by tumour cells, in which the substance is transformed into protoporphyrin IX, which exhibits a very strong and characteristic fluorescence in the red wavelength region. This procedure can be used, e.g. for the detection of early bronchial or bladder cancer and for the demarcation of the extension of a tumour for assuring a radical tumour treatment.
Background for the invention
Since long it is known that certain agent that are intravenously injected can be accumulated in tumours. If photo sensitizing agents are utilized such a tumour marking can be used in two ways, for diagnostics thorugh the fluorescence of the agent and for tumour treatment by photodynamic techniques, where the photo- excited agent transfers its energy to oxygen molecules, that make
a transition from their normal triplet state to an aggressive singlet state. The release of singlet oxygen selectively in the tumour causes tissue necrosis. During the last few years the commercially available agent Photofrin (Quadra Logic Technologies, Vancouver, Canada) has been injected and used clinically for the double purpose just described. Experimentally it has been found, that the selectivity for a tumour over surrounding tissue seldom exceeds a factor of 2-3, which limits the power of the method both for diagnostics and for therapy. It is possible to increase the discrimination for diagnostics by using also the autofluorescence of the tissue in the blue-green spectral region. This is utilized in procedures and equipment described in the Swedish patent # 455646 (Fluorescence device) and the European patent application # 0411104 (Improvements in diagnosis by means of fluorescent light emission from tissue). Also other agents, such as phtalocyanines, chlorines, purpurines and bensopo hyrin exhibit an accumulation in tumour tissue, but also for these the selectivity is limited, which reduces their value. Normal sensitizers have the difficulty that also the skin generally is sensitized, calling for the patient to be protected from strong day light for a period of about 4 weeks.
Description of the invention
The present invention is based on our observation, in animal experiments as well as clinically, that the agent δ-amino levulinic acid is an extremely efficient fluorescence marker for tumours. This is true for intravenous injection as well as for superficial treatment with ALA unguent direct on the tumour. The small non-fluorescing ALA molecules penetrate the cell membrane of malignant tumour cells and undergo in the mitochondria parts of the haem cycle, leading to the formation of protoporphyrin IX. Fluorescence can be induced by illumination by radiation from, e.g. a pulsed laser. This can be a dye laser pumped by a nitrogen laser. An especially suitable excitation wavelength is 405 nm, where porphyrins have their absorption maximum. The radiation can be conducted to the tissue through a quartz fibre, which can also be used to collect fluorescence light and to conduct it to a
spectral analyzer. This can be a spectrometer equipped with a diode-array detector or a more simple instrument based on optical filters. Tumours are marked by the occurrence of a fluorescence peak at 635 nm and a weaker peak at 710 nm. A demarcation ratio (the ratio between the specific fluorescence signal in a tumour and the signal in close-lying normal tissue) of about 10 or better can be obtained using this naturally occurring molecule in the body. It has no known side effects. A strong tumour demarcation is obtained after intra-venous injection if the investigation is performed about 1/2 hour after injection. ALA can also be prepared in an unguent base (about 20 % ALA) and be spread over superficial tumours and be kept in position using an occlusion bandage. The fluorescence investigation is performed 2-8 hours after the application.
Apart from exhibiting an increase in the specific red fluorescence, a decrease in the blue-green fluorescence from endogenous chromophores is observed in tumours. An especially effective form of diagnostics for ultraviolet or violet excitation is to evaluate the background-free fluorescence at 635 nm and divide it by the fluorescence intensity at about 470 nm. Such a ratio is dimensionless and therefore the signal will be independent of the measurement distance, the angle of incidence for the light, and fluctuations in the excitation and detection equipment.
The procedure given in this invention can also be used in combination with an imaging cancer detection system, which has been described in the Swedish patent # 455646. A simultaneous recording of images in three different fluorescence colours is then made (635 nm, 600 nm and 470 nm). An image intensifier, which is activated in synchronism with the exciting light source, is used for the recording. A new, generalized and tumour enhancing image is obtained by subtracting the 600 nm image (background) from the 635 nm image, followed by a division of the result by the blue image, pixel by pixel.
Cancer detection with fluorescence techiques using ALA is illustrated in Figure 1, where 1 indicates the excitation source, 2 indicates optical components to bring the exciting light to the tissue, 3 indicates the tissue that has been exposed to ALA, 4 indicates optical components for conducting the fluorescence light to a wavelength-dividing system 5 placed in front of the detection system 6. Finally, 7 indicates a signal processing system for tumour recognition based on the fluorescence properties.
The system proposed enables strongly improved possibilities for early detection of malignant tumours, also small tumours in the hollow organs of the body, and for clear demarcation of diseased tissue from surrounding normal tissue. No side effects occur.
The efficiency for this form of cancer detection is illustrated by the following three non-restricting examples.
Example 1.
A patient with basal cell carcinoma of the forehead was investigated with the method described in this invention. 6 hours before the fluorescence investigation the tumour areas were exposed to an unguent containing ALA in hydrochloric form (Porphyrin Products, Inc., Logan, Utah; catalogue # A 167). The unguent also covered a 10 mm wide zone of normal tissue. The fluorescence detection equipment used, as an excitation source, a Laser Science model VSL 337 pulsed nitrogen laser, pumping a Laser Science dye laser operating at a wavelength of 405 nm. The radiation from the dye laser was focussed by a lens into a quartz fibre. The other end of this fibre was put in contact with the tissue. Fluorescence light was collected through the same fibre, and after passing a dichroic beam splitter, used for injecting the laser light into the fibre, the fluorescence light was focussed on the entrance slit of a Jobin-Yvon 0.25 m grating monochromator equipped with an EG&G Optical Multichannel Analyser (OMA) HI system. Huorescence spectra from a basal cell cancer tumour and
from close-lying, ALA-exposed normal skin are shown in Figure 2. Using the intensity of the characteristic red fluorescence a demarcation ratio of 50 between the tumour and the normal tissue is obtained. If instead the ratio between the red and the blue- green fluorescence is used a demarcation ratio of 150 is obtained.
Example 2.
A patient with widespread local metastases of breast cancer was investigated with the same equipment and the same procedure as was described in Example 1. In Figure 3 fluorescence recordings 6 hours after application of the ALA unguent are shown for a skin area with lymphangitic metastatic breast cancer growth and for a close-lying normal skin area, also exposed to the ALA unguent. In this case the demarcation ratio is 13 and 25, respectively.
Example 3.
An experimental animal tumour model (human adenocarcinoma induced in the muscle of Wistar/Furth rats) was investigated with the method described in the present invention. The rat was intravenously injected with ALA hydrochloride in saline at a concentration of 30 mg/kg bodyweight. The rat was killed 30 minutes after the injection and the exposed tumour and the surrounding muscle were investigated using the same fluorescence set-up as was described in Example 1. Fluorescence signals from tumour and surrounding muscle are shown in Figure 4. A demarcation ratio of 9 is obtained using the characteristic red fluorescence. In this particular case, a division by the blue fluorescence intensity does not yield any further demarcation, for the excitation wavelength chosen, but still the advantages of recording a dimensionless quantity, as discussed above, are retained.
Figure captions
Figure 1. Schematic arrangment for fluorescence detection with ALA.
Figure 2. Fluorescence recordings for basal cell carcinoma and normal tissue for a patient that was exposed to an ALA unguent in the tumour area.
Figure 3. Fluorescence recordings for metastasis of breast cancer and normal tissue for a patient that was exposed to an ALA unguent in the tumour area.
Figure 4- Fluorescence recordings for an experimental animal tumour and surrounding normal tissue after intravenous injection of ALA.
Claims
1. A method for the detection of malignant tumours by fluorescence characterized by a substance, δ- aminolevulinic acid, which is supplied to the tissue and that the ratio between the specific peak of fluorescence intensity in red and in the blue-green is used to enhance the detection of tumours in investigations where the excitation is made in the ultra violet, violet or blue spectral range.
2. The method as recited in claim 1, characterized in that the applied substance is δ-aminolevulinic acid hydro-chloride.
3. The method as recited in claim 1, characterized in that the substance is a salve which is applied on to the suspicious malignant tissue.
4. The method as recited in claim 3, characterized in that the fluorescense examination is performed between 2 and 8 hours after the salve is applied.
5. The method as recited in claim 1, characterized in that the substance is intravenously administered.
6. The method as recited in claim 5, characterized in that the substance is injected with a concentration of 0.5 to 50 mg per kilo body weight.
7. The method as recited in claim 5, characterized in that the fluorescence examination is performed between 15 minutes to 2 hours after the injection.
8. The method as recited in claim 1, characterized in that the fluorescence examination is performed with laser excitation.
9. The method as recited in claim 1, characterized in that the fluorescence examination is performed with a conventional light source.
10. The method as recited in claim 1, characterized in that the fluorescence examination is performed with a fibre optic point monitoring system with excitation and detection through the same fibre.
11. The method as recited in claim 1, characterized in that the fluorescence examination is performed with a multi-color imaging system with subsequent data handling system.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| SE9103837-2 | 1991-12-21 | ||
| SE9103837A SE9103837L (en) | 1991-12-21 | 1991-12-21 | FLUORESCENSE DIAGNOSIS OF CANCER USING DELTA AMINOLEVULIC ACID |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| WO1993013403A1 true WO1993013403A1 (en) | 1993-07-08 |
Family
ID=20384715
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/SE1992/000879 WO1993013403A1 (en) | 1991-12-21 | 1992-12-18 | FLUORESCENCE DIAGNOSTICS OF CANCER USING δ-AMINO LEVULINIC ACID |
Country Status (3)
| Country | Link |
|---|---|
| EP (1) | EP0588994A1 (en) |
| SE (1) | SE9103837L (en) |
| WO (1) | WO1993013403A1 (en) |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997011636A2 (en) * | 1995-09-26 | 1997-04-03 | Karl Storz Gmbh & Co. | Apparatus for photodynamic diagnosis |
| EP0783867A1 (en) * | 1995-12-21 | 1997-07-16 | Unilever Plc | Device for the identification of acne, micromedones, and bacteria on human skin |
| DE19618963A1 (en) * | 1996-05-10 | 1997-11-13 | Herbert Dr Stepp | Endoscope auxiliary system determining fluorescent or re-emitted light on surfaces |
| WO1998009155A1 (en) * | 1996-08-27 | 1998-03-05 | Medeikonos Ab | Method for detecting cancer on skin of humans and mammals and arrangement for performing the method |
| EP0845457A1 (en) * | 1995-07-12 | 1998-06-03 | Mitsubishi Chemical Corporation | 2,2-dideutero-5-aminolevulinic acid |
| US6989140B2 (en) | 2001-12-21 | 2006-01-24 | Threshold Pharmaceuticals, Inc. | Methods for cancer imaging |
| EP1742038A1 (en) * | 2005-07-06 | 2007-01-10 | Academisch Medisch Centrum bij de Universiteit van Amsterdam | Device and method for determining the concentration of a substance |
| US7289205B2 (en) | 2003-09-19 | 2007-10-30 | The General Hospital Corporation | Fluorescence polarization imaging devices and methods |
| US7627363B2 (en) | 2003-03-18 | 2009-12-01 | The General Hospital Corporation | Polarized light imaging devices and methods |
| CZ301399B6 (en) * | 2004-09-03 | 2010-02-17 | Stabilizing dispersive base for photodynamic diagnostics and therapy | |
| CN103608662A (en) * | 2011-06-29 | 2014-02-26 | 京都府公立大学法人 | Tumor site identification device and method |
| US10920260B2 (en) | 2008-08-15 | 2021-02-16 | Erasmus University Medical Center Rotterdam | Methods and devices for assessment of mitochondrial function |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SE455646B (en) * | 1984-10-22 | 1988-07-25 | Radians Innova Ab | FLUORESCENT DEVICE |
| GB2203831A (en) * | 1986-07-07 | 1988-10-26 | Academy Of Applied Sciences | Diagnosis of malignant tumours by fluorescence |
| WO1990010219A1 (en) * | 1989-02-22 | 1990-09-07 | Spectraphos Ab | Improvements in diagnosis by means of fluorescent light emission from tissue |
| WO1991001727A2 (en) * | 1989-07-28 | 1991-02-21 | Queen's University At Kingston | Method of detection and treatment of malignant and non-malignant lesions by photochemotherapy |
-
1991
- 1991-12-21 SE SE9103837A patent/SE9103837L/en not_active Application Discontinuation
-
1992
- 1992-12-18 WO PCT/SE1992/000879 patent/WO1993013403A1/en not_active Application Discontinuation
-
1993
- 1993-07-19 EP EP19930901458 patent/EP0588994A1/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| SE455646B (en) * | 1984-10-22 | 1988-07-25 | Radians Innova Ab | FLUORESCENT DEVICE |
| GB2203831A (en) * | 1986-07-07 | 1988-10-26 | Academy Of Applied Sciences | Diagnosis of malignant tumours by fluorescence |
| WO1990010219A1 (en) * | 1989-02-22 | 1990-09-07 | Spectraphos Ab | Improvements in diagnosis by means of fluorescent light emission from tissue |
| WO1991001727A2 (en) * | 1989-07-28 | 1991-02-21 | Queen's University At Kingston | Method of detection and treatment of malignant and non-malignant lesions by photochemotherapy |
Non-Patent Citations (1)
| Title |
|---|
| Applied Optics, Volume 28, No. 12, June 1989, G.C. TANG et al., "Pulsed and cw laser fluorescense spectra from cancerous, normal, and chemically treated normal human breast and lung tissues", see the whole document. * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0845457A4 (en) * | 1995-07-12 | 1998-10-07 | Mitsubishi Chem Corp | 2,2-dideutero-5-aminolevulinic acid |
| US5945564A (en) * | 1995-07-12 | 1999-08-31 | Mitsubishi Chemical Corporation | 2,2-dideutero-5-aminolevulinic acid |
| EP0845457A1 (en) * | 1995-07-12 | 1998-06-03 | Mitsubishi Chemical Corporation | 2,2-dideutero-5-aminolevulinic acid |
| WO1997011636A3 (en) * | 1995-09-26 | 1997-06-05 | Storz Karl Gmbh & Co | Apparatus for photodynamic diagnosis |
| WO1997011636A2 (en) * | 1995-09-26 | 1997-04-03 | Karl Storz Gmbh & Co. | Apparatus for photodynamic diagnosis |
| EP0783867A1 (en) * | 1995-12-21 | 1997-07-16 | Unilever Plc | Device for the identification of acne, micromedones, and bacteria on human skin |
| US5760407A (en) * | 1995-12-21 | 1998-06-02 | Elizabeth Arden Co., Division Of Conopco, Inc. | Device for the identification of acne, microcomedones, and bacteria on human skin |
| DE19618963C5 (en) * | 1996-05-10 | 2004-01-22 | Stepp, Herbert, Dr. | Endoscope auxiliary device and method for endoscopic detection of fluorescent light using the same |
| DE19618963C2 (en) * | 1996-05-10 | 2000-01-27 | Herbert Stepp | Endoscope auxiliary device and method for endoscopic detection of fluorescent light using the same |
| DE19618963A1 (en) * | 1996-05-10 | 1997-11-13 | Herbert Dr Stepp | Endoscope auxiliary system determining fluorescent or re-emitted light on surfaces |
| WO1998009155A1 (en) * | 1996-08-27 | 1998-03-05 | Medeikonos Ab | Method for detecting cancer on skin of humans and mammals and arrangement for performing the method |
| AU718207B2 (en) * | 1996-08-27 | 2000-04-13 | Medeikonos Ab | Method for detecting cancer on skin of humans and mammals and arrangement for performing the method |
| US6421455B1 (en) | 1996-08-27 | 2002-07-16 | Medeikonos Ab | Method for detecting cancer on skin of humans and mammals and arrangement for performing the method |
| US6989140B2 (en) | 2001-12-21 | 2006-01-24 | Threshold Pharmaceuticals, Inc. | Methods for cancer imaging |
| US7627363B2 (en) | 2003-03-18 | 2009-12-01 | The General Hospital Corporation | Polarized light imaging devices and methods |
| US7289205B2 (en) | 2003-09-19 | 2007-10-30 | The General Hospital Corporation | Fluorescence polarization imaging devices and methods |
| US7564550B2 (en) | 2003-09-19 | 2009-07-21 | The General Hospital Corporation | Fluorescence polarization imaging devices and methods |
| US8139211B2 (en) | 2003-09-19 | 2012-03-20 | The General Hospital Corporation | Fluorescence polarization imaging device and method |
| CZ301399B6 (en) * | 2004-09-03 | 2010-02-17 | Stabilizing dispersive base for photodynamic diagnostics and therapy | |
| WO2007004873A1 (en) * | 2005-07-06 | 2007-01-11 | Academisch Medisch Centrum Bij De Universiteit Van Amsterdam | Device and method for determining the concentration of a substance |
| EP1742038A1 (en) * | 2005-07-06 | 2007-01-10 | Academisch Medisch Centrum bij de Universiteit van Amsterdam | Device and method for determining the concentration of a substance |
| EP2273255A1 (en) * | 2005-07-06 | 2011-01-12 | Academisch Medisch Centrum bij de Universiteit van Amsterdam | Device and method for determining the concentration of a substance |
| US8008038B2 (en) | 2005-07-06 | 2011-08-30 | Academisch Medicsh Centrum bij de Universiteit van Amsterdam | Methods for determining oxygen concentration with protoporphyrin IX |
| US10920260B2 (en) | 2008-08-15 | 2021-02-16 | Erasmus University Medical Center Rotterdam | Methods and devices for assessment of mitochondrial function |
| CN103608662A (en) * | 2011-06-29 | 2014-02-26 | 京都府公立大学法人 | Tumor site identification device and method |
| CN103608662B (en) * | 2011-06-29 | 2016-09-28 | 京都府公立大学法人 | The identification device of tumor locus and recognition methods |
Also Published As
| Publication number | Publication date |
|---|---|
| SE9103837D0 (en) | 1991-12-21 |
| SE9103837L (en) | 1993-06-22 |
| EP0588994A1 (en) | 1994-03-30 |
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